WO1998018930A2 - Streptococcus pneumoniae antigens and vaccines - Google Patents

Streptococcus pneumoniae antigens and vaccines Download PDF

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Publication number
WO1998018930A2
WO1998018930A2 PCT/US1997/019422 US9719422W WO9818930A2 WO 1998018930 A2 WO1998018930 A2 WO 1998018930A2 US 9719422 W US9719422 W US 9719422W WO 9818930 A2 WO9818930 A2 WO 9818930A2
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Prior art keywords
seq
amino acid
polypeptide
polypeptides
nucleotide
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PCT/US1997/019422
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French (fr)
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WO1998018930A3 (en
Inventor
Charles A. Kunsch
Gil H. Choi
L. Sydnor Johnson
Alex Hromockyj
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Human Genome Sciences, Inc.
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Application filed by Human Genome Sciences, Inc. filed Critical Human Genome Sciences, Inc.
Priority to DK97946850.1T priority Critical patent/DK0942983T4/en
Priority to DE69737125.5T priority patent/DE69737125T3/en
Priority to EP97946850.1A priority patent/EP0942983B2/en
Priority to JP52066798A priority patent/JP4469026B2/en
Priority to CA002269663A priority patent/CA2269663A1/en
Priority to AU51945/98A priority patent/AU5194598A/en
Priority to ES97946850.1T priority patent/ES2277362T5/en
Publication of WO1998018930A2 publication Critical patent/WO1998018930A2/en
Publication of WO1998018930A3 publication Critical patent/WO1998018930A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/315—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci
    • C07K14/3156—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci from Streptococcus pneumoniae (Pneumococcus)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/315—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies

Definitions

  • the present invention relates to novel Streptococcus pneumoniae antigens for the detection of Streptococcus and for the prevention or attenuation of disease caused by Streptococcus.
  • the invention further relates to isolated nucleic acid molecules encoding antigenic polypeptides of S. pneumoniae.
  • Antigenic polypeptides are also provided, as are vectors, host cells and recombinant methods for producing the same.
  • the invention additionally relates to diagnostic methods for detecting Streptococcus gene expression.
  • Streptococcus pneumoniae has been one of the most extensively studied microorganisms since its first isolation in 1881. It was the object of many investigations that led to important scientific discoveries. In 1928, Griffith observed that when heat-killed encapsulated pneumococci and live strains constitutively lacking any capsule were concomitantly injected into mice, the nonencapsulated could be converted into encapsulated pneumococci with the same capsular type as the heat-killed strain. Years later, the nature of this "transforming principle," or carrier of genetic information, was shown to be
  • pneumococcus is responsible for the death of a large number of children under the age of 5 years from pneumococcal pneumonia.
  • the incidence of pneumococcal disease is highest in infants under 2 years of age and in people over 60 years of age.
  • Pneumococci are the second most frequent cause (after Haemophilus influenzae type b) of bacterial meningitis and otitis media in children.
  • H. influenzae type b conjugate vaccines for H. influenzae type b
  • pneumococcal meningitis is likely to become increasingly prominent.
  • S. pneumoniae is the most important etiologic agent of community-acquired pneumonia in adults and is the second most common cause of bacterial meningitis behind Neisseria meningitidis.
  • the antibiotic generally prescribed to treat S. pneumoniae is benzylpenicillin, although resistance to this and to other antibiotics is found occasionally.
  • Pneumococcal resistance to penicillin results from mutations in its penicillin-binding proteins.
  • Erythromycin or clindamycin can be used to treat pneumonia in patients hypersensitive to penicillin, but resistant strains to these drugs exist.
  • Broad spectrum antibiotics e.g., the tetracyclines
  • S. pneumoniae Various proteins have been suggested to be involved in the pathogenicity of S. pneumoniae, however, only a few of them have actually been confirmed as virulence factors.
  • Pneumococci produce an IgAl protease that might interfere with host defense at mucosal surfaces. (Kornfield, S.J., et al. , Rev. Inf. Dis. 5:521-534 ( 1981 ).
  • S. pneumoniae also produces neuraminidase, an enzyme that may facilitate attachment to epithelial cells by cleaving sialic acid from the host glycolipids and gangliosides.
  • neuraminidase Partially purified neuraminidase was observed to induce meningitis-like symptoms in mice; however, the reliability of this finding has been questioned because the neuraminidase preparations used were probably contaminated with cell wall products.
  • Other pneumococcal proteins besides neuraminidase are involved in the adhesion of pneumococci to epithelial and endothelial cells. These pneumococcal proteins have as yet not been identified. Recently, Cundell et al. , reported that peptide permeases can modulate pneumococcal adherence to epithelial and endothelial cells.
  • pneumoniae genome consists of one circular, covalently closed, double- stranded DNA and a collection of so-called variable accessory elements, such as prophages, plasmids, transposons and the like. Most physical characteristics and almost all of the genes of S. pneumoniae are unknown. Among the few that have been identified, most have not been physically mapped or characterized in detail. Only a few genes of this organism have been sequenced. (See, for instance current versions of GENBANK and other nucleic acid databases, and references that relate to the genome of S. pneumoniae such as those set out elsewhere herein.) Identification of in v ⁇ -expressed, and broadly protective, antigens of 5. pneumoniae has remained elusive.
  • the present invention provides isolated nucleic acid molecules comprising polynucleotides encoding the S. pneumoniae polypeptides described in Table 1 and having the amino acid sequences shown as SEQ LD NO:2, SEQ ID NO:4, SEQ ID NO:6, and so on through SEQ ID NO:226.
  • one aspect of the invention provides isolated nucleic acid molecules comprising polynucleotides having a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence encoding any of the amino acid sequences of the polypeptides shown in Table 1 ; and (b) a nucleotide sequence complementary to any of the nucleotide sequences in (a).
  • nucleic acid embodiments of the invention relate to isolated nucleic acid molecules comprising polynucleotides which encode the amino acid sequences of epitope-bearing portions of an S. pneumoniae polypeptide having an amino acid sequence in (a) above.
  • the present invention also relates to recombinant vectors, which include the isolated nucleic acid molecules of the present invention, and to host cells containing the recombinant vectors, as well as to methods of making such vectors and host cells and for using these vectors for the production of S. pneumoniae polypeptides or peptides by recombinant techniques.
  • the invention further provides isolated S. pneumoniae polypeptides having an amino acid sequence selected from the group consisting of an amino acid sequence of any of the polypeptides described in Table 1.
  • polypeptides of the present invention also include polypeptides having an amino acid sequence with at least 70% similarity, and more preferably at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% similarity to those described in Table 1 , as well as polypeptides having an amino acid sequence at least 70% identical, more preferably at least 75% identical, and still more preferably 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to those above; as well as isolated nucleic acid molecules encoding such polypeptides.
  • the present invention further provides a vaccine, preferably a multi-component vaccine comprising one or more of the S. pneumoniae polynucleotides or polypeptides described in Table 1 , or fragments thereof, together with a pharmaceutically acceptable diluent, carrier, or excipient, wherein the S. pneumoniae polypeptide(s) are present in an amount effective to elicit an immune response to members of the Streptococcus genus in an animal.
  • a vaccine preferably a multi-component vaccine comprising one or more of the S. pneumoniae polynucleotides or polypeptides described in Table 1 , or fragments thereof, together with a pharmaceutically acceptable diluent, carrier, or excipient, wherein the S. pneumoniae polypeptide(s) are present in an amount effective to elicit an immune response to members of the Streptococcus genus in an animal.
  • a vaccine preferably a multi-component vaccine comprising one or more of the S. pneumoniae polynu
  • pneumoniae polypeptides of the present invention may further be combined with one or more immunogens of one or more other streptococcal or non-streptococcal organisms to produce a multi-component vaccine intended to elicit an immunological response against members of the Streptococcus genus and, optionally, one or more non-streptococcal organisms.
  • the vaccines of the present invention can be administered in a DNA form, e.g., "naked" DNA, wherein the DNA encodes one or more streptococcal polypeptides and, optionally, one or more polypeptides of a non-streptococcal organism.
  • the DNA encoding one or more polypeptides may be constructed such that these polypeptides are expressed fusion proteins.
  • the vaccines of the present invention may also be administered as a component of a genetically engineered organism.
  • a genetically engineered organism which expresses one or more S. pneumoniae polypeptides may be administered to an animal.
  • such a genetically engineered organism may contain one or more S. pneumoniae polypeptides of the present invention intracellularly, on its cell surface, or in its periplasmic space.
  • such a genetically engineered organism may secrete one or more S. pneumoniae polypeptides.
  • the vaccines of the present invention may be co-administered to an animal with an immune system modulator (e.g., CD86 and GM-CSF).
  • an immune system modulator e.g., CD86 and GM-CSF
  • the invention also provides a method of inducing an immunological response in an animal to one or more members of the Streptococcus genus, preferrably one or more isolates of the S. pneumoniae genus, comprising administering to the animal a vaccine as described above.
  • the invention further provides a method of inducing a protective immune response in an animal, sufficient to prevent or attenuate an infection by members of the Streptococcus genus, preferrably at least S. pneumoniae, comprising administering to the animal a composition comprising one or more of the polynucleotides or polypeptides described in Table 1 , or fragments thereof. Further, these polypeptides, or fragments thereof, may be conjugated to another immunogen and/or administered in admixture with an adjuvant.
  • the invention further relates to antibodies elicited in an animal by the administration of one or more S. pneumoniae polypeptides of the present invention and t methods for producing such antibodies.
  • the invention also provides diagnostic methods for detecting the expression of genes of members of the Streptococcus genus in an animal.
  • One such method involves assaying for the expression of a gene encoding S. pneumoniae peptides in a sample from an animal. This expression may be assayed either directly (e.g. , by assaying polypeptide levels using antibodies elicited in response to amino acid sequences described in Table 1) or indirectly (e.g. , by assaying for antibodies having specificity for amino acid sequences described in Table 1).
  • An example of such a method involves the use of the polymerase chain reaction (PCR) to amplify and detect Streptococcus nucleic acid sequences.
  • PCR polymerase chain reaction
  • the present invention also relates to nucleic acid probes having all or part of a nucleotide sequence described in Table 1 (shown as SEQ ID NO: l, SEQ ID NO:3, SEQ ID NO:5, and so on through SEQ ID NO:225) which are capable of hybridizing under stringent conditions to Streptococcus nucleic acids.
  • the invention further relates to a method of detecting one or more Streptococcus nucleic acids in a biological sample obtained from an animal, said one or more nucleic acids encoding Streptococcus polypeptides, comprising: (a) contacting the sample with one or more of the above-described nucleic acid probes, under conditions such that hybridization occurs, and (b) detecting hybridization of said one or more probes to the Streptococcus nucleic acid present in the biological sample.
  • the invention also includes immunoassays, including an immunoassay for detecting Streptococcus, preferrably at least isolates of the S.
  • pneumoniae genus comprising incubation of a sample (which is suspected of being infected with Streptococcus) with a probe antibody directed against an antigen/epitope of S. pneumoniae, to be detected under conditions allowing the formation of an antigen-antibody complex; and detecting the antigen-antibody complex which contains the probe antibody.
  • An immunoassay for the detection of antibodies which are directed against a Streptococcus antigen comprising the incubation of a sample (containing antibodies from a mammal suspected of being infected with Streptococcus) with a probe polypeptide including an epitope of S. pneumoniae , under conditions that allow the formation of antigen-antibody complexes which contain the probe epitope containing antigen.
  • kits are those for: investigating samples for the presence of polynucleotides derived from Streptococcus which comprise a polynucleotide probe including a nucleotide sequence selected from Table 1 or a fragment thereof of approximately 15 or more nucleotides, in an appropriate container; analyzing the samples for the presence of antibodies directed against a Streptococcus antigen made up of a polypeptide which contains a S. pneumoniae epitope present in the polypeptide, in a suitable container; and analyzing samples for the presence of Streptococcus antigens made up of an anti-5. pneumoniae antibody, in a suitable container.
  • the present invention relates to recombinant antigenic S. pneumoniae polypeptides and fragments thereof.
  • the invention also relates to methods for using these polypeptides to produce immunological responses and to confer immunological protection to disease caused by members of the genus Streptococcus, at least isolates of the S. pneumoniae genus.
  • the invention further relates to nucleic acid sequences which encode antigenic S. pneumoniae polypeptides and to methods for detecting S. pneumoniae nucleic acids and polypeptides in biological samples.
  • the invention also relates to S. pneumoniae -specific antibodies and methods for detecting such antibodies produced in a host animal.
  • pathogenic agent means an agent which causes a disease state or affliction in an animal. Included within this definition, for examples, are bacteria, protozoans, fungi, viruses and metazoan parasites which either produce a disease state or render an animal infected with such an organism susceptible to a disease state (e.g., a secondary infection). Further included are species and strains of the genus Streptococcus which produce disease states in animals.
  • the term "organism” means any living biological system, including viruses, regardless of whether it is a pathogenic agent.
  • Streptococcus means any species or strain of bacteria which is members of the genus Streptococcus. Such species and strains are known to those of skill in the art, and include those that are pathogenic and those that are not.
  • one or more S. pneumoniae polypeptides of the present invention means polypeptides comprising the amino acid sequence of one or more, of the S. pneumoniae polypeptides described in Table 1 and disclosed as SEQ ID NO:2, SEQ ID NO:4, SEQ ED NO:6, and so on through SEQ ID NO:226. These polypeptides may be expressed as fusion proteins wherein the 5. pneumoniae polypeptides of the present invention are linked to additional amino acid sequences which may be of streptococcal or non- streptococcal origin. This phrase further includes polypeptide comprising fragments of the S. pneumoniae polypeptides of the present invention.
  • Table 1 provides information describing 1 13 open reading frames (ORFs) which encode potentially antigenic polypeptides of S. pneumoniae of the present invention.
  • ORFs open reading frames
  • the table lists the ORF identifier which consists of the letters SP, which denote S. pneumoniae, followed immediately by a three digit numeric code, which arbitrarily number the potentially antigenic polypeptides of S. pneumoniae of the present invention and the nucleotide or amino acid sequence of each ORF and encoded polypeptide.
  • the table further correlates the ORF identifier with a sequence identification number (SEQ ID NO:).
  • SEQ ID NO: sequence identification number
  • the actual nucleotide or amino acid sequence of each ORF identifier is also shown in the Sequence Listing under the corresponding SEQ ID NO.
  • SPI 26 refers to both the nucleotide and amino acid sequences of S. pneumoniae polypeptide number 126 of the present invention.
  • SP126 correlates with the nucleotide sequence shown as SEQ ED NO:223 and with the amino acid sequence shown as SEQ ID NO:224 as is described in Table 1.
  • each "ORF” The open reading frame within each "ORF" begins with the second nucleotide shown.
  • the first codon for each nucleotide sequence shown is bases 2-4, the second 5-7, the third 8-10, and so on.
  • Table 2 lists the antigenic epitopes present in each of the S. pneumoniae polypeptides described in Table 1 as predicted by the inventors. Each S . pneumoniae polypeptide shown in Table 1 has one or more antigenic epitopes described in Table 2. It will be appreciated that depending on the analytical criteria used to predict antigenic determinants, the exact address of the determinant may vary slightly. The exact location of the antigenic determinant may shift by about 1 to 5 residues, more likely 1 to 2 residues, depending on the criteria used. Thus, the first antigenic determinant described in Table 2 ,
  • “Lys-1 to Ile-10" of SP001 represents a peptide comprising the lysine at position 1 in SEQ ED NO:2 through and including the isoleucine at position 10 in SEQ ID NO:2, but may include more or fewer residues than those 10. It will also be appreciated that, generally speaking, amino acids can be added to either terminus of a peptide or polypeptide containing an antigenic epitope without affecting its activity, whereas removing residues from a peptide or polypeptide containing only the antigenic determinant is much more likely to destroy activity. It will be appreciated that the residues and locations shown described in Table 2 correspond to the amino acid sequences for each ORF shown in Table 1 and in the Sequence Listing.
  • Table 3 shows PCR primers designed by the inventors for the amplification of polynucleotides encoding polypeptides of the present invention according to the method of Example 1.
  • PCR primer design is routine in the art and those shown in Table 3 are provided merely for the convenience of the skilled artisan. It will be appreciated that others can be used with equal success.
  • the present invention provides a select number of ORFs from those presented in the fragments of the S. pneumoniae genome which may prove useful for the generation of a protective immune response.
  • the sequenced S . pneumoniae genomic DNA was obtained from a sub-cultured isolate of S . pneumoniae Strain 7/87 14.8.91 , which has been deposited at the American
  • Type Culture Collection as a convenience to those of skill in the art.
  • the 5. pneumoniae isolate was deposited on October 10, 1996 at the ATCC, 12301 Park Lawn Drive, Rockville, Maryland 20852, and given accession number 55840.
  • a genomic library constructed from DNA isolated from the S. pneumoniae isolate was also deposited at the ATCC on October 1 1 , 1996 and given ATCC E ⁇ eposit No. 97755.
  • a more complete listing of the sequence obtained from the S. pneumoniae genome may be found in co-pending U.S . Provisional Application Serial No. 60/029,960, filed 10/31/96, inco ⁇ orated herein by reference in its entirety.
  • Some ORFs contained in the subset of fragments of the S. pneumoniae genome disclosed herein were derived through the use of a number of screening criteria detailed below.
  • the selected ORFs do not consist of complete ORFs.
  • a polypeptide representing a complete ORF may be the closest approximation of a protein native to an organism, it is not always preferred to express a complete ORF in a heterologous system. It may be challenging to express and purify a highly hydrophobic protein by common laboratory methods.
  • the polypeptide vaccine candidates described herein may have been modified slightly to simplify the production of recombinant protein. For example, nucleotide sequences which encode highly hydrophobic domains, such as those found at the amino terminal signal sequence, have been excluded from some constructs used for in vitro expression of the polypeptides.
  • polypeptide which represents a truncated or modified ORF may be used as an antigen.
  • Type I signal sequence An amino terminal type I signal sequence generally directs a nascent protein across the plasma and outer membranes to the exterior of the bacterial cell.
  • Type IV signal sequence The type IV signal sequence is an example of the several types of functional signal sequences which exist in addition to the type I signal sequence detailed above.
  • type IV signal sequence Although functionally related, the type IV signal sequence possesses a unique set of biochemical and physical attributes (Strom, M. S. and Lory, S., J. Bacteriol. 174:7345-735 1 ( 1992)). These are typically six to eight amino acids with a net basic charge followed by an additional sixteen to thirty primarily hydrophobic residues.
  • the cleavage site of a type IV signal sequence is typically after the initial six to eight amino acids at the extreme amino terminus.
  • type EV signal sequences generally contain a phenylalanine residue at the +1 site relative to the cleavage site. 3.
  • Lipoprotein Studies of the cleavage sites of twenty-six bacterial lipoprotein precursors has allowed the definition of a consensus amino acid sequence for lipoprotein cleavage. Nearly three-fourths of the bacterial lipoprotein precursors examined contained the sequence L-(A,S)-(G,A)-C at positions -3 to +1, relative to the point of cleavage (Hayashi, S. and Wu, H . C, J. Bioenerg. Biomembr. 22:451-471 ( 1990)).
  • LPXTG motif It has been experimentally determined that most anchored proteins found on the surface of gram-positive bacteria possess a highly conserved carboxy terminal sequence. More than fifty such proteins from organisms such as S. pyogenes, S. mutans, E. faecalis, S. pneumoniae, and others, have been identified based on their extracellular location and carboxy terminal amino acid sequence (Fischetti, V. A., ASM News 62:405-410 ( 1996)). The conserved region consists of six charged amino acids at the extreme carboxy terminus coupled to 15-20 hydrophobic amino acids presumed to function as a transmembrane domain. Immediately adjacent to the transmembrane domain is a six amino acid sequence conserved in nearly all proteins examined.
  • the amino acid sequence of this region is L-P-X-T-G-X, where X is any amino acid.
  • An algorithm for selecting antigenic and immunogenic S. pneumoniae polypeptides including the foregoing criteria was developed. Use of the algorithm by the inventors to select immunologically useful S. pneumoniae polypeptides resulted in the selection of a number of the disclosed ORFs. Polypeptides comprising the polypeptides identified in this group may be produced by techniques standard in the art and as further described herein.
  • the present invention provides isolated nucleic acid molecules comprising polynucleotides encoding the S. pneumoniae polypeptides having the amino acid sequences described in Table 1 and shown as SEQ ED NO:2,
  • SEQ ID NO:4 SEQ ID NO:6, and so on through SEQ ED NO:226, which were determined by sequencing the genome of S. pneumoniae and selected as putative immunogens.
  • nucleotide sequences determined by sequencing a DNA molecule herein were determined using an automated DNA sequencer (such as the Model 373 from Applied Biosystems, Inc.), and all amino acid sequences of polypeptides encoded by DNA molecules determined herein were predicted by translation of DNA sequences determined as above. Therefore, as is known in the art for any DNA sequence determined by this automated approach, any nucleotide sequence determined herein may contain some errors. Nucleotide sequences determined by automation are typically at least about 90% identical, more typically at least about 95% to at least about 99.9% identical to the actual nucleotide sequence of the sequenced DNA molecule. The actual sequence can be more precisely determined by other approaches including manual DNA sequencing methods well known in the art.
  • a single insertion or deletion in a determined nucleotide sequence compared to the actual sequence will cause a frame shift in translation of the nucleotide sequence such that the predicted amino acid sequence encoded by a determined nucleotide sequence will be completely different from the amino acid sequence actually encoded by the sequenced DNA molecule, beginning at the point of such an insertion or deletion.
  • nucleotide sequence set forth herein is presented as a sequence of deoxyribonucleotides (abbreviated A, G , C and T).
  • nucleic acid molecule or polynucleotide a sequence of deoxyribonucleotides
  • RNA molecule or polynucleotide the corresponding sequence of ribonucleotides (A, G, C and U), where each thymidine deoxyribonucleotide (T) in the specified deoxyribonucleotide sequence is replaced by the ribonucleotide uridine (U).
  • RNA molecule having a sequence described in Table 1 set forth using deoxyribonucleotide abbreviations is intended to indicate an RNA molecule having a sequence in which each deoxyribonucleotide A, G or C described in Table 1 has been replaced by the corresponding ribonucleotide A, G or C, and each deoxyribonucleotide T has been replaced by a ribonucleotide U.
  • Nucleic acid molecules of the present invention may be in the form of RNA, such as mRNA, or in the form of DNA, including, for instance, cDNA and genomic DNA obtained by cloning or produced synthetically.
  • the DNA may be double-stranded or single-stranded.
  • Single-stranded DNA or RNA may be the coding / Strand, also known as the sense strand, or it may be the non-coding strand, also referred to as the anti-sense strand.
  • isolated nucleic acid molecule(s) is intended a nucleic acid molecule, DNA or RNA, which has been removed from its native environment.
  • recombinant DNA molecules contained in a vector are considered isolated for the purposes of the present invention.
  • Further examples of isolated DNA molecules include recombinant DNA molecules maintained in heterologous host cells or purified (partially or substantially) DNA molecules in solution.
  • Isolated RNA molecules include in vivo or in.vitro RNA transcripts of the DNA molecules of the present invention. Isolated nucleic acid molecules according to the present invention further include such molecules produced synthetically.
  • Isolated nucleic acid molecules of the present invention include DNA molecules comprising a nucleotide sequence described in Table 1 and shown as SEQ ED NO: l , SEQ ID NO:3, SEQ ID NO:5, and so on through SEQ ED
  • the genetic code is well known in the art. Thus, it would be routine for one skilled in the art to generate such degenerate variants.
  • the invention also provides nucleic acid molecules having sequences complementary to any one of those described in Table 1. Such isolated molecules, particularly DNA molecules, are useful as probes for detecting expression of Streptococcal genes, for instance, by Northern blot analysis or the polymerase chain reaction (PCR).
  • the present invention is further directed to fragments of the isolated nucleic acid molecules described herein.
  • a fragment of an isolated nucleic acid molecule having a nucleotide sequence described in Table 1 is intended fragments at least about 15 nt, and more preferably at least about 17 nt, still more preferably at least about 20 nt, and even more preferably, at least about 25 nt in length which are useful as diagnostic probes and primers as discussed herein.
  • larger fragments 50-100 nt in length are also useful according to the present invention as are fragments corresponding to most, if not all, of a nucleotide sequence described in Table 1.
  • fragments which include 20 or more contiguous bases of a nucleotide sequence as described in Table 1. Since the nucleotide sequences identified in Table 1 are provided as SEQ ED NO: l , SEQ ID NO:3, SEQ ID NO:5, and so on through SEQ ED NO:225, generating such DNA fragments would be routine to the skilled artisan. For example, such fragments could be generated synthetically.
  • nucleic acid fragments of the present invention also include nucleic acid molecules comprising nucleotide sequences encoding epitope-bearing portions of the S. pneumoniae polypeptides identified in Table 1.
  • Such nucleic acid fragments of the present invention include, for example, nucleotide sequences encoding polypeptide fragments comprising from about the amino terminal residue to about the carboxy terminal residue of each fragment shown in Table 2.
  • the above referred to polypeptide fragments are antigenic regions of the S. pneumoniae polypeptides identified in Table 1.
  • the invention provides isolated nucleic acid molecules comprising polynucleotides which hybridize under stringent hybridization conditions to a portion of a polynucleotide in a nucleic acid molecule of the invention described above, for instance, a nucleic acid sequence identified in Table 1.
  • stringent hybridization conditions is intended overnight incubation at 42°C in a solution comprising: 50% formamide, 5x SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5x
  • polynucleotides which hybridize to a "portion" of a polynucleotide is intended polynucleotides (either DNA or RNA) which hybridize to at least about 15 nucleotides (nt), and more preferably at least about 17 nt, still more preferably at least about 20 nt, and even more preferably about 25-70 nt of the reference polynucleotide. These are useful as diagnostic probes and primers as discussed above and in more detail below.
  • polynucleotides hybridizing to a larger portion of the reference polynucleotide are also useful as probes according to the present invention, as are polynucleotides corresponding to most, if not all, of a nucleotide sequence as identified in Table 1.
  • a portion of a polynucleotide of "at least 20 nt in length,” for example, is intended 20 or more contiguous nucleotides from the nucleotide sequence of the reference polynucleotide (e.g., a nucleotide sequences as described in Table 1).
  • portions are useful diagnostically either as probes according to conventional DNA hybridization techniques or as primers for amplification of a target sequence by PCR, as described in the literature (for instance, in Molecular Cloning, A Laboratory Manual, 2nd. edition, Sambrook, J., Fritsch, E. F. and Maniatis, T., eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. ( 1989), the entire disclosure of which is hereby inco ⁇ orated herein by reference).
  • nucleic acid sequences encoding the S. pneumoniae polypeptides of the present invention are identified in Table 1 and provided as SEQ ED NO: 1 , SEQ ID NO:3, SEQ ED NO:5, and so on through SEQ ED NO:225, generating polynucleotides which hybridize to portions of these sequences would be routine to the skilled artisan.
  • the hybridizing polynucleotides of the present invention could be generated synthetically according to known techniques.
  • nucleic acid molecules of the present invention which encode S. pneumoniae polypeptides of the present invention may include, but are not limited to those encoding the amino acid sequences of the polypeptides by themselves; and additional coding sequences which code for additional amino acids, such as those which provide additional functionalities.
  • the sequences encoding these polypeptides may be fused to a marker sequence, such as a sequence encoding a peptide which facilitates purification of the fused polypeptide.
  • the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (Qiagen, Inc.), among others, many of which are commercially available. As described by Gentz and colleagues (Proc. Natl. Acad. Sci. USA 86:821-824 ( 1989)), for instance, hexa-histidine provides for convenient purification of the resulting fusion protein.
  • the present invention also includes genetic fusions wherein the S . pneumoniae nucleic acid sequences coding sequences identified in Table 1 are linked to additional nucleic acid sequences to produce fusion proteins.
  • These fusion proteins may include epitopes of streptococcal or non-streptococcal origin designed to produce proteins having enhanced immunogenicity.
  • the fusion proteins of the present invention may contain antigenic determinants known to provide helper T-cell stimulation, peptides encoding sites for post-translational modifications which enhance immunogenicity (e.g. , acylation), peptides which facilitate purification (e.g., histidine "tag”), or amino acid sequences which target the fusion protein to a desired location (e.g.
  • the invention includes polypeptides shown in Table 1 with, and without an N-termainal methionine.
  • the present invention thus includes nucleic acid molecules and sequences which encode fusion proteins comprising one or more S. pneumoniae polypeptides of the present invention fused to an amino acid sequence which allows for post-translational modification to enhance immunogenicity.
  • This post-translational modification may occur either in vitro or when the fusion protein is expressed in vivo in a host cell.
  • An example of such a modification is the introduction of an amino acid sequence which results in the attachment of a lipid moiety.
  • the present invention includes genetic fusions wherein a S. pneumoniae nucleic acid sequence identified in Table 1 is linked to a nucleotide sequence encoding another amino acid sequence.
  • These other amino acid sequences may be of streptococcal origin (e.g. , another sequence selected from Table 1) or non-streptococcal origin.
  • the present invention further relates to variants of the nucleic acid molecules of the present invention, which encode portions, analogs or derivatives of the S. pneumoniae polypeptides described in Table 1.
  • Variants may occur naturally, such as a natural allelic variant.
  • allelic variant is intended one of several alternate forms of a gene occupying a given locus on a chromosome of an organism (Genes II, Lewin, B., ed., John Wiley & Sons, New York ( 1985)).
  • Non-naturally occurring variants may be produced using art-known mutagenesis techniques.
  • Such variants include those produced by nucleotide substitutions, deletions or additions.
  • the substitutions, deletions or additions may involve one or more nucleotides.
  • These variants may be altered in coding regions, non-coding regions, or both. Alterations in the coding regions may produce conservative or non-conservative amino acid substitutions, deletions or additions. Especially preferred among these are silent substitutions, additions and deletions, which do not alter the properties and activities of the S. pneumoniae polypeptides disclosed herein or portions thereof.
  • Silent substitution are most likely to be made in non-epitopic regions. Guidance regarding those regions containing epitopes is provided herein, for example, in
  • nucleotide sequence encoding any of the amino acid sequences of the polypeptides identified in Table 1; and (b) a nucleotide sequence complementary to any of the nucleotide sequences in (a) above.
  • nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding the subject S . pneumoniae polypeptide.
  • a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence.
  • These mutations of the reference sequence may occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence.
  • nucleotides within some of the nucleic acid sequences shown in Table 1 were ambiguous upon sequencing. Completely unknown sequences are shown as an "N". Other unresolved nucleotides are known to be either a purine, shown as “R”, or a pyrimidine, shown as "Y”. Accordingly, when determining identity between two nucleotide sequences, identity is met where any nucleotide, including an "R”, “Y” or “N", is found in a test sequence and at the corresponding position in the referece sequence (from Table 1). Likewise, an A, G or "R” in a test sequence is identical to an "R” in the reference sequence; and a T, C or "Y” in a test sequence is identical to a "Y” in the reference sequence.
  • nucleotide sequence described in Table 1 can be determined conventionally using known computer programs such as the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wl 5371 1). Bestfit uses the local homology algorithm of Smith and Waterman (Advances in Applied Mathematics 2:482-489 ( 1981)), to find the best segment of homology between two sequences.
  • the parameters are set, of course, such that the percentage of identity is calculated over the full length of the reference nucleotide sequence and that gaps in homology of up to 5% of the total number of nucleotides in the reference sequence are allowed.
  • nucleic acid molecules at least 90%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequences described in Table 1.
  • nucleic acid molecules for instance, as a hybridization probe or a polymerase chain reaction (PCR) primer.
  • Uses of the nucleic acid molecules of the present invention include, inter alia, ( 1) isolating Streptococcal genes or allelic variants thereof from either a genomic or cDNA library and (2) Northern Blot or PCR analysis for detecting Streptococcal mRNA expression.
  • nucleic acid molecules having a sequence at least 90%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence identified in Table 1 will encode the same polypeptide.
  • degenerate variants of these nucleotide sequences all encode the same polypeptide, this will be clear to the skilled artisan even without performing the above described comparison assay.
  • antigenic epitopes have been identified which contain as few as six amino acids (see Harlow, et al, Antibodies: A Laboratory Manual, 2nd Ed.; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York (1988), page 76), in instances where a polypeptide has multiple antigenic epitopes the alteration of several amino acid residues would often not be expected to eliminate all of the antigenic epitopes of that polypeptide. This is especially so when the alterations are in regions believed to not constitute antigenic epitopes.
  • Vectors and Host Cells The present invention also relates to vectors which include the isolated
  • DNA molecules of the present invention are DNA molecules of the present invention, host cells which are genetically engineered with the recombinant vectors, and the production of S. pneumoniae polypeptides or fragments thereof by recombinant techniques.
  • Recombinant constructs may be introduced into host cells using well known techniques such as infection, transduction, transfection, transvection, electroporation and transformation.
  • the vector may be, for example, a phage, plasmid, viral or retroviral vector. Retroviral vectors may be replication competent or replication defective. In the latter case, viral propagation generally will occur only in complementing host cells.
  • the polynucleotides may be joined to a vector containing a selectable marker for propagation in a host.
  • a plasmid vector is introduced in a precipitate, such as a calcium phosphate precipitate, or in a complex with a charged lipid.
  • the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells.
  • Appropriate frans-acting factors may be supplied by the host, supplied by a complementing vector or supplied by the vector itself upon introduction into the host.
  • the vectors provide for specific expression, which may be inducible and/or cell type-specific.
  • Expression vectors useful in the present invention include chromosomal-, episomal- and virus-derived vectors, e.g. , vectors derived from bacterial plasmids, bacteriophage, yeast episomes, yeast chromosomal elements, viruses such as baculoviruses, papova viruses, vaccinia viruses, adenoviruses, fowl pox viruses, pseudorabies viruses and retroviruses, and vectors derived from combinations thereof, such as cosmids and phagemids.
  • vectors derived from bacterial plasmids, bacteriophage, yeast episomes, yeast chromosomal elements, viruses such as baculoviruses, papova viruses, vaccinia viruses, adenoviruses, fowl pox viruses, pseudorabies viruses and retroviruses and vectors derived from combinations thereof, such as cosmids and phagemids.
  • the DNA insert should be operatively linked to an appropriate promoter, such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few.
  • an appropriate promoter such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few.
  • Other suitable promoters will be known to the skilled artisan.
  • the expression constructs will further contain sites for transcription initiation, termination and, in the transcribed region, a ribosome binding site for translation.
  • the coding portion of the mature transcripts expressed by the constructs will preferably include a translation initiating site at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated.
  • the expression vectors will preferably include at least one selectable marker.
  • markers include dihydrofolate reductase or neomycin resistance for eukaryotic cell culture and tetracycline or ampicillin resistance genes for culturing in E. coli and other bacteria.
  • Representative examples of appropriate hosts include, but are not limited to, bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS and Bowes melanoma cells; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.
  • vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from Qiagen; pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNHl ⁇ a, pNH18A, pNH46A available from Stratagene; pET series of vectors available from Novagen; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRET5 available from Pharmacia.
  • eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXTl and pSG available from Stratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia.
  • Other suitable vectors will be readily apparent to the skilled artisan.
  • known bacterial promoters suitable for use in the present invention include the E. coli lacl and lacZ promoters, the T3 and T7 promoters, the gpt promoter, the lambda PR and PL promoters and the trp promoter.
  • Suitable eukaryotic promoters include the CMV immediate early promoter, the HSV thymidine kinase promoter, the early and late SV40 promoters, the promoters of retroviral LTRs, such as those of the Rous sarcoma virus (RSV), and metallothionein promoters, such as the mouse metallothionein-I promoter.
  • Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection or other methods. Such methods are described in many standard laboratory manuals (for example. Davis, et al, Basic Methods In Molecular Biology (1986)).
  • Enhancers are cw-acting elements of DNA, usually about from 10 to 300 bp that act to increase transcriptional activity of a promoter in a given host cell-type.
  • enhancers include the SV40 enhancer, which is located on the late side of the replication origin at bp 100 to 270, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.
  • secretion signals may be inco ⁇ orated into the expressed polypeptide.
  • the signals may be endogenous to the polypeptide or they may be heterologous signals.
  • the polypeptide may be expressed in a modified form, such as a fusion protein, and may include not only secretion signals, but also additional heterologous functional regions. For instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence in the host cell, during purification, or during subsequent handling and storage. Also, peptide moieties may be added to the polypeptide to facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to polypeptides to engender secretion or excretion, to improve stability and to facilitate purification, among others, are familiar and routine techniques in the art.
  • a preferred fusion protein comprises a heterologous region from immunoglobulin that is useful to solubilize proteins.
  • EP-A-O 464 533 (Canadian counte ⁇ art 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobin molecules together with another human protein or part thereof.
  • the Fc part in a fusion protein is thoroughly advantageous for use in therapy and diagnosis and thus results, for example, in improved pharmacokinetic properties (EP-A 0232 262).
  • Fc portion proves to be a hindrance to use in therapy and diagnosis, for example when the fusion protein is to be used as antigen for immunizations.
  • human proteins such as, hEL5-receptor has been fused with Fc portions for the pu ⁇ ose of high-throughput screening assays to identify antagonists of hIL-5. See Bennett, D. et al, J. Molec. Recogn. 5:52-58 ( 1995) and Johanson, K. et al, J. Biol Chem. 270 (76,1:9459-9471 (1995). The S.
  • pneumoniae polypeptides can be recovered and purified from recombinant cell cultures by well-known methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography, lectin chromatography and high performance liquid chromatography ("HPLC”) is employed for purification.
  • Polypeptides of the present invention include naturally purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect and mammalian cells.
  • the invention further provides isolated polypeptides having the amino acid sequences described in Table 1. and shown as SEQ ED NO:2, SEQ ED NO:4, SEQ ED NO:6, and so on through SEQ ID NO:226, and peptides or polypeptides comprising portions of the above polypeptides.
  • the terms "peptide” and “oligopeptide” are considered synonymous (as is commonly recognized) and each term can be used interchangeably as the context requires to indicate a chain of at least two amino acids coupled by peptidyl linkages.
  • the word "polypeptide” is used herein for chains containing more than ten amino acid residues. All oligopeptide and polypeptide formulas or sequences herein are written from left to right and in the direction from amino terminus to carboxy terminus.
  • S. pneumoniae polypeptides described in Table 1 can be varied without significantly effecting the antigenicity of the polypeptides. If such differences in sequence are contemplated, it should be remembered that there will be critical areas on the polypeptide which determine antigenicity. In general, it is possible to replace residues which do not form part of an antigenic epitope without significantly effecting the antigenicity of a polypeptide. Guidance for such alterations is given in Table 2 wherein epitopes for each polypeptide is delineated.
  • polypeptides of the present invention are preferably provided in an isolated form.
  • isolated polypeptide is intended a polypeptide removed from its native environment.
  • a polypeptide produced and/or contained within a recombinant host cell is considered isolated for pu ⁇ oses of the present invention.
  • isolated polypeptide is a polypeptide that has been purified, partially or substantially, from a recombinant host cell.
  • recombinantly produced versions of the S. pneumoniae polypeptides described in Table 1 can be substantially purified by the one-step method described by Smith and Johnson (Gene 67:31-40 ( 1988)).
  • polypeptides of the present invention include: (a) an amino acid sequence of any of the polypeptides described in Table 1 ; and (b) an amino acid sequence of an epitope-bearing portion of any one of the polypeptides of (a); as well as polypeptides with at least 70% similarity, and more preferably at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% similarity to those described in (a) or (b) above, as well as polypeptides having an amino acid sequence at least 70% identical, more preferably at least 75% identical, and still more preferably 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to those above.
  • % similarity for two polypeptides is intended a similarity score produced by comparing the amino acid sequences of the two polypeptides using the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science
  • polypeptide having an amino acid sequence at least, for example,
  • a reference amino acid sequence of a S. pneumoniae polypeptide is intended that the amino acid sequence of the polypeptide is identical to the reference sequence except that the polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the reference amino acid sequence.
  • up to 5% of the amino acid residues in the reference sequence may be deleted or substituted with another amino acid, or a number of amino acids up to 5% of the total amino acid residues in the reference sequence may be inserted into the reference sequence.
  • These alterations of the reference sequence may occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence.
  • amino acid sequences shown in Table 1 may have on or more "X" residues. "X” represents unknown. Thus, for pu ⁇ oses of defining identity, if any amino acid is present at the same position in a reference amino acid sequence (shown in Table 1) where an X is shown, the two sequences are identical at that position.
  • any particular polypeptide is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to, for instance, an amino acid sequence shown in Table 1 , can be determined conventionally using known computer programs such the Bestfit program
  • polypeptides of the present invention can also be used to raise polyclonal and monoclonal antibodies, which are useful in assays for detecting Streptococcal protein expression.
  • the invention provides peptides and polypeptides comprising epitope-bearing portions of the S. pneumoniae polypeptides of the invention.
  • These epitopes are immunogenic or antigenic epitopes of the polypeptides of the invention.
  • An "immunogenic epitope" is defined as a part of a protein that elicits an antibody response when the whole protein or polypeptide is the immunogen. These immunogenic epitopes are believed to be confined to a few loci on the molecule.
  • an antigenic determinant or "antigenic epitope.”
  • the number of immunogenic epitopes of a protein generally is less than the number of antigenic epitopes (Gey sen, et al, Proc. Natl Acad. Sci. USA 81:3998- 4002 ( 1983)).
  • Predicted antigenic epitopes are shown in Table 2, below. As to the selection of peptides or polypeptides bearing an antigenic epitope (i.e.
  • Peptides capable of eliciting protein-reactive sera are frequently represented in the primary sequence of a protein, can be characterized by a set of simple chemical rules, and are confined neither to immunodominant regions of intact proteins (i.e. , immunogenic epitopes) nor to the amino or carboxyl terminals. Peptides that are extremely hydrophobic and those of six or fewer residues generally are ineffective at inducing antibodies that bind to the mimicked protein; longer, peptides, especially those containing proline residues, usually are effective (Sutcliffe, et al , supra, p. 661).
  • Antigenic epitope-bearing peptides and polypeptides of the invention are therefore useful to raise antibodies, including monoclonal antibodies, that bind specifically to a polypeptide of the invention.
  • hybridomas obtained by fusion of spleen cells from donors immunized with an antigen epitope-bearing peptide generally secrete antibody reactive with the native protein (Sutcliffe, et al , supra, p. 663).
  • the antibodies raised by antigenic epitope-bearing peptides or polypeptides are useful to detect the mimicked protein, and antibodies to different peptides may be used for tracking the fate of various regions of a protein precursor which undergoes post-translational processing.
  • the peptides and anti-peptide antibodies may be used in a variety of qualitative or quantitative assays for the mimicked protein, for instance in competition assays since it has been shown that even short peptides (e.g., about 9 amino acids) can bind and displace the larger peptides in immunoprecipitation assays (for instance, Wilson, et al, Cell 37:767-778 ( 1984) p. 777).
  • the anti-peptide antibodies of the invention also are useful for purification of the mimicked protein, for instance, by adso ⁇ tion chromatography using methods well known in the art.
  • Antigenic epitope-bearing peptides and polypeptides of the invention designed according to the above guidelines preferably contain a sequence of at least seven, more preferably at least nine and most preferably between about 15 to about 30 amino acids contained within the amino acid sequence of a polypeptide of the invention.
  • peptides or polypeptides comprising a larger portion of an amino acid sequence of a polypeptide of the invention, containing about 30 to about 50 amino acids, or any length up to and including the entire amino acid sequence of a polypeptide of the invention also are considered epitope-bearing peptides or polypeptides of the invention and also are useful for inducing antibodies that react with the mimicked protein.
  • the amino acid sequence of the epitope-bearing peptide is selected to provide substantial solubility in aqueous solvents (i.e. , the sequence includes relatively hydrophilic residues and highly hydrophobic sequences are preferably avoided); and sequences containing proline residues are particularly preferred.
  • Non-limiting examples of antigenic polypeptides or peptides that can be used to generate Streptococcal-specific antibodies include portions of the amino acid sequences identified in Table 1. More specifically, Table 2 discloses antigenic fragments of polypeptides of the present invention, which antigenic fragments comprise amino acid sequences from about the first amino acid residues indicated to about the last amino acid residue indicated for each fragment. The polypeptide fragments disclosed in Table 2 are believed to be antigenic regions of the S. pneumoniae polypeptides described in Table 1. Thus the invention further includes isolated peptides and polypeptides comprising an amino acid sequence of an epitope shown in Table 2 and polynucleotides encoding said polypeptides.
  • the epitope-bearing peptides and polypeptides of the invention may be produced by any conventional means for making peptides or polypeptides including recombinant means using nucleic acid molecules of the invention.
  • an epitope-bearing amino acid sequence of the present invention may be fused to a larger polypeptide which acts as a carrier during recombinant production and purification, as well as during immunization to produce anti-peptide antibodies.
  • Epitope-bearing peptides also may be synthesized using known methods of chemical synthesis.
  • Houghten has described a simple method for synthesis of large numbers of peptides, such as 10-20 mg of 248 different 13 residue peptides representing single amino acid variants of a segment of the HA1 polypeptide which were prepared and characterized (by ELISA-type binding studies) in less than four weeks (Houghten, R. A. Proc.
  • Epitope-bearing peptides and polypeptides of the invention are used to induce antibodies according to methods well known in the art (for instance, Sutcliffe, et al., supra; Wilson, et al, supra; Chow, M., et al, Proc. Natl. Acad. Sci. USA 82:910-914; and Bittle, F. J., et al, J. Gen. Virol 66:2347-2354 ( 1985)).
  • animals may be immunized with free peptide; however, anti-peptide antibody titer may be boosted by coupling of the peptide to a macromolecular carrier, such as keyhole limpet hemacyanin (KLH) or tetanus toxoid.
  • KLH keyhole limpet hemacyanin
  • peptides containing cysteine may be coupled to carrier using a linker such as m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), while other peptides may be coupled to carrier using a more general linking agent such as glutaraldehyde.
  • Animals such as rabbits, rats and mice are immunized with either free or carrier-coupled peptides, for instance, by intraperitoneal and/or intradermal injection of emulsions containing about 100 ⁇ g peptide or carrier protein and Freund's adjuvant. Several booster injections may be needed, for instance, at intervals of about two weeks, to provide a useful titer of anti-peptide antibody which can be detected, for example, by ELISA assay using free peptide adsorbed to a solid surface.
  • the titer of anti-peptide antibodies in serum from an immunized animal may be increased by selection of anti-peptide antibodies, for instance, by adso ⁇ tion to the peptide on a solid support and elution of the selected antibodies according to methods well known in the art.
  • Immunogenic epitope-bearing peptides of the invention i.e., those parts of a protein that elicit an antibody response when the whole protein is the immunogen, are identified according to methods known in the art. For instance, Geysen, et al , supra, discloses a procedure for rapid concurrent synthesis on solid supports of hundreds of peptides of sufficient purity to react in an enzyme-linked immunosorbent assay. Interaction of synthesized peptides with antibodies is then easily detected without removing them from the support. In this manner a peptide bearing an immunogenic epitope of a desired protein may be identified routinely by one of ordinary skill in the art.
  • the immunologically important epitope in the coat protein of foot-and-mouth disease virus was located by Geysen et al. supra with a resolution of seven amino acids by synthesis of an overlapping set of all 208 possible hexapeptides covering the entire 213 amino acid sequence of the protein. Then, a complete replacement set of peptides in which all 20 amino acids were substituted in turn at every position within the epitope were synthesized, and the particular amino acids conferring specificity for the reaction with antibody were determined.
  • peptide analogs of the epitope-bearing peptides of the invention can be made routinely by this method.
  • U.S. Patent No. 4,708,781 to Geysen (1987) further describes this method of identifying a peptide bearing an immunogenic epitope of a desired protein.
  • U.S. Patent No. 5, 194,392, to Geysen ( 1990) describes a general method of detecting or determining the sequence of monomers (amino acids or other compounds) which is a topological equivalent of the epitope (i.e. , a "mimotope") which is complementary to a particular paratope (antigen binding site) of an antibody of interest. More generally, U.S. Patent No. 4,433,092, also to Geysen ( 1989), describes a method of detecting or determining a sequence of monomers which is a topographical equivalent of a ligand which is complementary to the ligand binding site of a particular receptor of interest. Similarly, U.S. Patent No. 5,480,971 to Houghten, R.
  • A. et al. discloses linear C -C -alkyl peralkylated oligopeptides and sets and libraries of such peptides, as well as methods for using such oligopeptide sets and libraries for determining the sequence of a peralkylated oligopeptide that preferentially binds to an acceptor molecule of interest.
  • non-peptide analogs of the epitope-bearing peptides of the invention also can be made routinely by these methods.
  • polypeptides of the present invention and the epitope-bearing fragments thereof described above can be combined with parts of the constant domain of immunoglobulins (IgG), resulting in chimeric polypeptides.
  • IgG immunoglobulins
  • These fusion proteins facilitate purification and show an increased half-life in vivo. This has been shown, e.g. , for chimeric proteins consisting of the first two domains of the human CD4-polypeptide and various domains of the constant regions of the heavy or light chains of mammalian immunoglobulins (EPA 0,394,827; Traunecker et al, Nature 337:84-86 ( 1988)).
  • Fusion proteins that have a disulfide-linked dimeric structure due to the IgG part can also be more efficient in binding and neutralizing other molecules than a monomeric S. pneumoniae polypeptide or fragment thereof alone (Fountoulakis et al, J. Biochem. 270:3958-3964 ( 1995)).
  • the present invention further relates to a method for assaying for
  • Streptococcal infection in an animal via detecting the expression of genes encoding Streptococcal polypeptides (e.g., the polypeptides described Table 1 ).
  • This method comprises analyzing tissue or body fluid from the animal for Streptococcus-specific antibodies or Streptococcal nucleic acids or proteins. Analysis of nucleic acid specific to Streptococcus can be done by PCR or hybridization techniques using nucleic acid sequences of the present invention as either hybridization probes or primers (cf. Molecular Cloning: A Laboratory Manual, second edition, edited by Sambrook, Fritsch, & Maniatis, Cold Spring Harbor Laboratory, 1989; Eremeeva et al, J. Clin.
  • Streptococcus has already been made, the present invention is useful for monitoring progression or regression of the disease state whereby patients exhibiting enhanced Streptococcus gene expression will experience a worse clinical outcome relative to patients expressing these gene(s) at a lower level.
  • assaying for Streptococcal infection in an animal via detection of genes encoding Streptococcal polypeptides is intended qualitatively or quantitatively measuring or estimating the level of one or more Streptococcus polypeptides or the level of nucleic acid encoding Streptococcus polypeptides in a first biological sample either directly (e.g. , by determining or estimating absolute protein level or nucleic level) or relatively (e.g. , by comparing to the
  • Streptococcus polypeptide level or mRNA level in a second biological sample may be undetectable if obtained from an animal which is not infected with Streptococcus.
  • the Streptococcus polypeptide level or nucleic acid level may be compared to a second sample obtained from either an animal infected with Streptococcus or the same animal from which the first sample was obtained but taken from that animal at a different time than the first.
  • a standard Streptococcus polypeptide level or nucleic acid level which corresponds to a particular stage of a Streptococcus infection is known, it can be used repeatedly as a standard for comparison.
  • biological sample any biological sample obtained from an animal, cell line, tissue culture, or other source which contains Streptococcus polypeptide, mRNA, or DNA.
  • Biological samples include body fluids (such as plasma and synovial fluid) which contain Streptococcus polypeptides, and muscle, skin, and cartilage tissues. Methods for obtaining tissue biopsies and body fluids are well known in the art. The present invention is useful for detecting diseases related to
  • Streptococcus infections in animals include monkeys, apes, cats, dogs, cows, pigs, mice, horses, rabbits and humans. Particularly preferred are humans.
  • Total RNA can be isolated from a biological sample using any suitable technique such as the single-step guanidinium-thiocyanate-phenol-chloroform method described in Chomczynski and Sacchi, Anal Biochem. 762: 156-159 ( 1987).
  • mRNA encoding Streptococcus polypeptides having sufficient homology to the nucleic acid sequences identified in Table 1 to allow for hybridization between complementary sequences are then assayed using any appropriate method. These include Northern blot analysis, S I nuclease mapping, the polymerase chain reaction (PCR), reverse transcription in combination with the polymerase chain reaction (RT-PCR), and reverse transcription in combination with the ligase chain reaction (RT-LCR).
  • PCR polymerase chain reaction
  • RT-PCR reverse transcription in combination with the polymerase chain reaction
  • RT-LCR reverse transcription in combination with the ligase chain reaction
  • RNA is prepared from a biological sample as described above.
  • an appropriate buffer such as glyoxal/dimethyl sulfoxide/sodium phosphate buffer
  • agarose gel electrophoresis is transferred onto a nitrocellulose filter.
  • the filter is prehybridized in a solution containing formamide, SSC,
  • pnuemoniae DNA sequence of the present invention is used as a template to synthesize labeled antisense DNA.
  • the antisense DNA can then be digested using an appropriate restriction endonuclease to generate further DNA probes of a desired length.
  • Such antisense probes are useful for visualizing protected bands corresponding to the target mRNA (i.e., mRNA encoding Streptococcus polypeptides).
  • levels of mRNA encoding Streptococcus polypeptides are assayed using the RT-PCR method described in Makino et al,
  • this method involves adding total RNA isolated from a biological sample in a reaction mixture containing a RT primer and appropriate buffer. After incubating for primer annealing, the mixture can be supplemented with a RT buffer, dNTPs, DTP, RNase inhibitor and reverse transcriptase. After incubation to achieve reverse transcription of the RNA, the RT products are then subject to PCR using labeled primers. Alternatively, rather than labeling the primers, a labeled dNTP can be included in the PCR reaction mixture.
  • PCR amplification can be performed in a DNA thermal cycler according to conventional techniques. After a suitable number of rounds to achieve amplification, the PCR reaction mixture is electrophoresed on a polyacrylamide gel. After drying the gel, the radioactivity of the appropriate bands (corresponding to the mRNA encoding the Streptococcus polypeptides)) is quantified using an imaging analyzer. RT and PCR reaction ingredients and conditions, reagent and gel concentrations, and labeling methods are well known in the art. Variations on the RT-PCR method will be apparent to the skilled artisan.
  • Assaying Streptococcus polypeptide levels in a biological sample can occur using any art-known method.
  • Preferred for assaying Streptococcus polypeptide levels in a biological sample are antibody-based techniques.
  • Streptococcus polypeptide expression in tissues can be studied with classical immunohistological methods.
  • the specific recognition is provided by the primary antibody (polyclonal or monoclonal) but the secondary detection system can utilize fluorescent, enzyme, or other conjugated secondary antibodies.
  • an immunohistological staining of tissue section for pathological examination is obtained. Tissues can also be extracted, e.g.
  • ELISA enzyme linked immunosorbent assay
  • RIA radioimmunoassay
  • a Streptococcus polypeptide-specific monoclonal antibodies can be used both as an immunoabsorbent and as an enzyme-labeled probe to detect and quantify a Streptococcus polypeptide.
  • the amount of a Streptococcus polypeptide present in the sample can be calculated by reference to the amount present in a standard preparation using a linear regression computer algorithm.
  • ELISA enzyme linked immunosorbent assay
  • RIA radioimmunoassay
  • two distinct specific monoclonal antibodies can be used to detect Streptococcus polypeptides in a body fluid.
  • one of the antibodies is used as the immunoabsorbent and the other as the enzyme-labeled probe.
  • the above techniques may be conducted essentially as a "one-step” or “two-step” assay.
  • the "one-step” assay involves contacting the Streptococcus polypeptide with immobilized antibody and, without washing, contacting the mixture with the labeled antibody.
  • the "two-step” assay involves washing before contacting the mixture with the labeled antibody.
  • Other conventional methods may also be employed as suitable. It is usually desirable to immobilize one component of the assay system on a support, thereby allowing other components of the system to be brought into contact with the component and readily removed from the sample.
  • Streptococcus polypeptide-specific antibodies for use in the present invention can be raised against an intact S. pneumoize polypeptide of the present invention or fragment thereof. These polypeptides and fragments may be administered to an animal (e.g. , rabbit or mouse) either with a carrier protein
  • albumin e.g., albumin
  • long enough e.g., at least about 25 amino acids
  • antibody As used herein, the term "antibody” (Ab) or “monoclonal antibody” (Mab) is meant to include intact molecules as well as antibody fragments (such as, for example, Fab and F(ab') fragments) which are capable of specifically binding to a St rep tococcus polypeptide.
  • Fab and F(ab') ⁇ fragments lack the Fc fragment of intact antibody, clear more rapidly from the circulation, and may have less non-specific tissue binding of an intact antibody (Wahl et al, J. Nucl.
  • the antibodies of the present invention may be prepared by any of a variety of methods.
  • the S. pneumoniae polypeptides identified in Table 1 can be administered to an animal in order to induce the production of sera containing polyclonal antibodies.
  • a preparation of a S. pneumoniae polypeptide of the present invention is prepared and purified to render it substantially free of natural contaminants.
  • Such a preparation is then introduced into an animal in order to produce polyclonal antisera of high specific activity.
  • the antibodies of the present invention are monoclonal antibodies.
  • Such monoclonal antibodies can be prepared using hybridoma technology (Kohler et al, Nature 256:495 ( 1975); Kohler et al, Eur. J. Immunol. 6:51 1 ( 1976); Kohler et al, Eur. J. Immunol 6:292 ( 1976); Hammerling et al, In: Monoclonal Antibodies and T-Cell Hybridomas,
  • Such procedures involve immunizing an animal (preferably a mouse) with a S. pneumoniae polypeptide antigen of the present invention.
  • Suitable cells can be recognized by their capacity to bind ⁇ -Streptococcus polypeptide antibody.
  • Such cells may be cultured in any suitable tissue culture medium; however, it is preferable to culture cells in Earle's modified Eagle's medium supplemented with 10% fetal bovine serum (inactivated at about 56°C), and supplemented with about 10 g/1 of nonessential amino acids, about 1 ,000 U/ml of penicillin, and about 100 ⁇ g/ml of streptomycin.
  • the splenocytes of such mice are extracted and fused with a suitable myeloma cell line.
  • a suitable myeloma cell line may be employed in accordance with the present invention; however, it is preferable to employ the parent myeloma cell line (SP O), available from the American Type
  • hybridoma cells obtained through such a selection are then assayed to identify clones which secrete antibodies capable of binding the Streptococcus polypeptide antigen administered to immunized animal.
  • additional antibodies capable of binding to Streptococcus polypeptide antigens may be produced in a two-step procedure through the use of anti-idiotypic antibodies.
  • Such a method makes use of the fact that antibodies are themselves antigens, and that, therefore, it is possible to obtain an antibody which binds to a second antibody.
  • Streptococcus polypeptide-specific antibodies are used to immunize an animal, preferably a mouse.
  • the splenocytes of such an animal are then used to produce hybridoma cells, and the hybridoma cells are screened to identify clones which produce an antibody whose ability to bind to the Streptococcus polypeptide-specific antibody can be blocked by a Streptococcus polypeptide antigen.
  • Such antibodies comprise anti-idiotypic antibodies to the Streptococcus polypeptide-specific antibody and can be used to immunize an animal to induce formation of further Streptococcus polypeptide-specific antibodies.
  • Fab and F(ab') and other fragments of the antibodies of the present invention may be used according to the methods disclosed herein.
  • Such fragments are typically produced by proteolytic cleavage, using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab') fragments).
  • enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab') fragments).
  • Streptococcus polypeptide-binding fragments can be produced through the application of recombinant DNA technology or through synthetic chemistry.
  • Antibodies to Streptococcus polypeptide antigens which are produced in humans, or are "humanized” (i.e. , non-immunogenic in a human) by recombinant or other technology.
  • Humanized antibodies may be produced, for example by replacing an immunogenic portion of an antibody with a corresponding, but non- immunogenic portion (i.e. , chimeric antibodies) (Robinson, R.R. et al , International Patent Publication PCT/US86/02269; Akira, K. et al , European Patent Application 184, 187; Taniguchi, M., European Patent Application 171 ,496; Morrison, S.L. et al , European Patent Application 173,494;
  • Suitable enzyme labels include, for example, those from the oxidase group, which catalyze the production of hydrogen peroxide by reacting with substrate.
  • Glucose oxidase is particularly preferred as it has good stability and its substrate (glucose) is readily available.
  • Activity of an oxidase label may be assayed by measuring the concentration of hydrogen peroxide formed by the enzyme-labeled antibody/substrate reaction.
  • radioisotopes such as iodine ( " I, I), carbon ( C), sulphur
  • suitable labels for the Streptococcus polypeptide-specific antibodies of the present invention are provided below.
  • suitable enzyme labels include malate dehydrogenase, staphylococcal nuclease, delta-5-steroid isomerase, yeast-alcohol dehydrogenase, alpha-glycerol phosphate dehydrogenase, triose phosphate isomerase, peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, beta-galactosidase, ribonuclease, urease, catalase, glucose-6-phosphate dehydrogenase, glucoamylase, and acetylcholine esterase.
  • suitable radioisotopic labels include H, In, " I, I,
  • Suitable non-radioactive isotopic labels include Gd,
  • fluorescent labels examples include an Eu label, a fluorescein label, an isothiocyanate label, a rhodamine label, a phycoerythrin label, a phycocyanin label, an allophycocyanin label, an o-phthaldehyde label, and a fluorescamine label.
  • Suitable toxin labels include diphtheria toxin, ricin, and cholera toxin.
  • chemiluminescent labels include a luminal label, an isoluminal label, an aromatic acridinium ester label, an imidazole label, an acridinium salt label, an oxalate ester label, a luciferin label, a luciferase label, and an aequorin label.
  • nuclear magnetic resonance contrasting agents include heavy metal nuclei such as Gd, Mn, and iron.
  • Typical techniques for binding the above-described labels to antibodies are provided by Kennedy et al , Clin. Chim. Acta 70: 1-31 ( 1976), and Schurs et al, Clin. Chim. Acta 87: 1-40 (1977). Coupling techniques mentioned in the latter are the glutaraldehyde method, the periodate method, the dimaleimide method, the m-maleimidobenzyl-N-hydroxy-succinimide ester method, all of which methods are inco ⁇ orated by reference herein.
  • the invention includes a diagnostic kit for use in screening serum containing antibodies specific against S. pneumoniae infection.
  • a kit may include an isolated S. pneumoniae antigen comprising an epitope which is specifically immunoreactive with at least one anti-5. pneumoniae antibody.
  • Such a kit also includes means for detecting the binding of said antibody to the antigen.
  • the kit may include a recombinantly produced or chemically synthesized peptide or polypeptide antigen. The peptide or polypeptide antigen may be attached to a solid support.
  • the detecting means of the above- described kit includes a solid support to which said peptide or polypeptide antigen is attached.
  • a kit may also include a non-attached reporter-labelled anti-human antibody.
  • binding of the antibody to the S . pneumoniae antigen can be detected by binding of the reporter labelled antibody to the anti-5. pneumoniae antibody.
  • the invention includes a method of detecting S. pneumoniae infection in a subject.
  • This detection method includes reacting a body fluid, preferrably serum, from the subject with an isolated S. pneumoniae antigen, and examining the antigen for the presence of bound antibody.
  • the method includes a polypeptide antigen attached to a solid support, and serum is reacted with the support. Subsequently, the support is reacted with a reporter-labelled anti-human antibody. The support is then examined for the presence of reporter-labelled antibody.
  • the solid surface reagent employed in the above assays and kits is prepared by known techniques for attaching protein material to solid support material, such as polymeric beads, dip sticks, 96-well plates or filter material.
  • attachment methods generally include non-specific adso ⁇ tion of the protein to the support or covalent attachment of the protein , typically through a free amine group, to a chemically reactive group on the solid support, such as an activated carboxyl, hydroxyl, or aldehyde group.
  • streptavidin coated plates can be used in conjunction with biotinylated antigen(s).
  • the present invention also provides vaccines comprising one or more polypeptides of the present invention.
  • Heterogeneity in the composition of a vaccine may be provided by combining S. pneumoniae polypeptides of the present invention.
  • Multi -component vaccines of this type are desirable because they are likely to be more effective in eliciting protective immune responses against multiple species and strains of the Streptococcus genus than single polypeptide vaccines.
  • a multi-component vaccine of the present invention may contain one or more, preferably 2 to about 20, more preferably 2 to about 15, and most preferably 3 to about 8, of the 5. pneumoniae pojypeptides identified in Table 1 , or fragments thereof.
  • Multi-component vaccines are known in the art to elicit antibody production to numerous immunogenic components. Decker, M. and Edwards, K., I. Infect. Dis. 774:S270-275 ( 1996).
  • a hepatitis B, diphtheria, tetanus, pertussis tetravalent vaccine has recently been demonstrated to elicit protective levels of antibodies in human infants against all four pathogenic agents. Aristegui, J. et al, Vaccine 75:7-9 ( 1997).
  • the present invention thus also includes multi-component vaccines.
  • These vaccines comprise more than one polypeptide, immunogen or antigen.
  • An example of such a multi-component vaccine would be a vaccine comprising more than one of the S. pneumoniae polypeptides described in Table 1.
  • a second example is a vaccine comprising one or more, for example 2 to 10, of the S. pneumoniae polypeptides identified in Table 1 and one or more, for example 2 to 10, additional polypeptides of either streptococcal or non-streptococcal origin.
  • a multi-component vaccine which confers protective immunity to both a Streptococcal infection and infection by another pathogenic agent is also within the scope of the invention.
  • the vaccines of the present invention are expected to elicit a protective immune response against infections caused by species and strains of Streptococcus other than strain ofS. pneumoniae deposited with that
  • whole cell and whole viral vaccines are produced recombinantly and involve the expression of one or more of the S. pneumoniae polypeptides described in Table 1.
  • the S. pneumoniae polypeptides of the present invention may be either secreted or localized intracellular, on the cell surface, or in the periplasmic space.
  • the S. pneumoniae polypeptides of the present invention may, for example, be localized in the viral envelope, on the surface of the capsid, or internally within the capsid.
  • Whole cells vaccines which employ cells expressing heterologous proteins are known in the art. See, e.g. , Robinson, K. et al, Nature Biotech.
  • a multi-component vaccine can also be prepared using techniques known in the art by combining one or more S.
  • pneumoniae polypeptides of the present invention or fragments thereof, with additional non-streptococcal components (e.g. , diphtheria toxin or tetanus toxin, and/or other compounds known to elicit an immune response).
  • additional non-streptococcal components e.g. , diphtheria toxin or tetanus toxin, and/or other compounds known to elicit an immune response.
  • Such vaccines are useful for eliciting protective immune responses to both members of the Streptococcus genus and non-streptococcal pathogenic agents.
  • the vaccines of the present invention also include DNA vaccines.
  • DNA vaccines are currently being developed for a number of infectious diseases. Boyer, J et al, Nat. Med. 3:526-532 (1997); reviewed in Spier, R., Vaccine 14: 1285-1288 (1996).
  • Such DNA vaccines contain a nucleotide sequence encoding one or more S. pneumoniae polypeptides of the present invention oriented in a manner that allows for expression of the subject polypeptide.
  • the direct administration of plasmid DNA encoding B. burgdorgeri OspA has been shown to elicit protective immunity in mice against borrelial challenge. Luke, C. et al, J. Infect. Dis. 775:91-97 ( 1997).
  • the present invention also relates to the administration of a vaccine which is co-administered with a molecule capable of modulating immune responses.
  • Kim, J. et al, Nature Biotech. 75:641-646 ( 1997) report the enhancement of immune responses produced by DNA immunizations when DNA sequences encoding molecules which stimulate the immune response are co-administered.
  • the vaccines of the present invention may be co-administered with either nucleic acids encoding immune modulators or the immune modulators themselves.
  • These immune modulators include granulocyte macrophage colony stimulating factor (GM-CSF) and CD86.
  • GM-CSF granulocyte macrophage colony stimulating factor
  • the vaccines of the present invention may be used to confer resistance to streptococcal infection by either passive or active immunization.
  • a vaccine of the present invention is administered to an animal to elicit a protective immune response which either prevents or attenuates a streptococcal infection.
  • the vaccines of the present invention are used to confer resistance to streptococcal infection through passive immunization, the vaccine is provided to a host animal (e.g. , human, dog, or mouse), and the antisera elicited by this antisera is recovered and directly provided to a recipient suspected of having an infection caused by a member of the Streptococcus genus.
  • antibodies, or fragments of antibodies, with toxin molecules provides an additional method for treating streptococcal infections when passive immunization is conducted.
  • antibodies, or fragments of antibodies capable of recognizing the 5. pneumoniae polypeptides disclosed herein, or fragments thereof, as well as other Streptococcus proteins, are labeled with toxin molecules prior to their administration to the patient.
  • toxin derivatized antibodies bind to Streptococcus cells, toxin moieties will be localized to these cells and will cause their death.
  • the present invention thus concerns and provides a means for preventing or attenuating a streptococcal infection resulting from organisms which have antigens that are recognized and bound by antisera produced in response to the polypeptides of the present invention.
  • a vaccine is said to prevent or attenuate a disease if its administration to an animal results either in the total or partial attenuation (i.e. , suppression) of a symptom or condition of the disease, or in the total or partial immunity of the animal to the disease.
  • the administration of the vaccine may be for either a "prophylactic" or "therapeutic" pu ⁇ ose.
  • the compound(s) When provided prophylactically, the compound(s) are provided in advance of any symptoms of streptococcal infection.
  • the prophylactic administration of the compound(s) serves to prevent or attenuate any subsequent infection.
  • the compound(s) When provided therapeutically, the compound(s) is provided upon or after the detection of symptoms which indicate that an animal may be infected with a member of the Streptococcus genus.
  • the therapeutic administration of the compound(s) serves to attenuate any actual infection.
  • the S. pneumoniae polypeptides, and fragments thereof, of the present invention may be provided either prior to the onset of infection (so as to prevent or attenuate an anticipated infection) or after the initiation of an actual infection.
  • polypeptides of the invention may be administered in pure form or may be coupled to a macromolecular carrier.
  • macromolecular carrier examples include proteins and carbohydrates.
  • Suitable proteins which may act as macromolecular carrier for enhancing the immunogenicity of the polypeptides of the present invention include keyhole limpet hemacyanin (KLH) tetanus toxoid, pertussis toxin, bovine serum albumin, and ovalbumin.
  • KLH keyhole limpet hemacyanin
  • a composition is said to be "pharmacologically acceptable” if its administration pan be tolerated by a recipient animal and is otherwise suitable for administration to that animal.
  • Such an agent is said to be administered in a "therapeutically effective amount” if the amount administered is physiologically significant.
  • An agent is physiologically significant if its presence results in a detectable change in the physiology of a recipient patient.
  • the vaccine of the present invention is administered as a pharmacologically acceptable compound
  • a pharmacologically acceptable compound varies with the animal to which it is administered.
  • a vaccine intended for human use will generally not be co-administered with Freund's adjuvant.
  • the level of purity of the S. pneumoniae polypeptides of the present invention will normally be higher when administered to a human than when administered to a non-human animal.
  • the vaccine of the present invention when provided to an animal, it may be in a composition which may contain salts, buffers, adjuvants, or other substances which are desirable for improving the efficacy of the composition.
  • Adjuvants are substances that can be used to specifically augment a specific immune response. These substances generally perform two functions: ( 1) they protect the antigen(s) from being rapidly catabolized after administration and (2) they nonspecifically stimulate immune responses.
  • Adjuvants can be loosely divided into several groups based upon their composition. These groups include oil adjuvants (for example, Freund's complete and incomplete), mineral salts (for example, AlK(SO ) AlNa(SO ⁇ , A1NH (SO ), silica, kaolin, and carbon), polynucleotides (for example, poly IC and poly AU acids), and certain natural substances (for example, wax D from Mycobacterium tuberculosis, as well as substances found in Corynebacterium parvum, or Bordetella pertussis, and members of the genus Brucella. Other substances useful as adjuvants are the saponins such as, for example, Quil A. (Superfos A/S, Denmark). Preferred adjuvants for use in the present invention include aluminum salts, such as
  • compositions of the present invention can be administered parenterally by injection, rapid infusion, nasopharyngeal abso ⁇ tion (intranasopharangeally), dermoabso ⁇ tion, or orally.
  • the compositions may alternatively be administered intramuscularly, or intravenously.
  • compositions for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions.
  • non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.
  • Carriers or occlusive dressings can be used to increase skin permeability and enhance antigen abso ⁇ tion.
  • Liquid dosage forms for oral administration may generally comprise a liposome solution containing the liquid dosage form.
  • Suitable forms for suspending liposomes include emulsions, suspensions, solutions, syrups, and elixirs containing inert diluents commonly used in the art, such as purified water.
  • such compositions can also include adjuvants, wetting agents, emulsifying and suspending agents, or sweetening, flavoring, or perfuming agents.
  • compositions of the present invention can also be administered in encapsulated form.
  • intranasal immunization of mice against Bordetella pertussis infection using vaccines encapsulated in biodegradable microsphere composed of poly(DL-lactide-co-glycolide) has been shown to stimulate protective immune responses.
  • Encapsulated vaccines of the present invention can be administered by a variety of routes including those involving contacting the vaccine with mucous membranes (e.g., intranasally, intracolonicly, intraduodenally). Many different techniques exist for the timing of the immunizations when a multiple administration regimen is utilized. It is possible to use the compositions of the invention more than once to increase the levels and diversities of expression of the immunoglobulin repertoire expressed by the immunized animal. Typically, if multiple immunizations are given, they will be given one to two months apart.
  • an "effective amount" of a therapeutic composition is one which is sufficient to achieve a desired biological effect.
  • the dosage needed to provide an effective amount of the composition will vary depending upon such factors as the animal's or human's age, condition, sex, and extent of disease, if any, and other variables which can be adjusted by pne of ordinary skill in the art.
  • the antigenic preparations of the invention can be administered by either single or multiple dosages of an effective amount.
  • Effective amounts of the compositions of the invention can vary from 0.01- 1 ,000 ⁇ g/ml per dose, more preferably 0.1-500 ⁇ g/ml per dose, and most preferably 10-300 ⁇ g/ml per dose.
  • the components of the pQElO plasmid are arranged such that the inserted DNA sequence encoding a polypeptide of the present invention expresses the polypeptide with the six His residues (i. e. , a "6 X His tag”)) covalently linked to the amino terminus.
  • the DNA sequences encoding the desired portions of the polypeptides of Table 1 are amplified using PCR oligonucleotide primers from either a DNA library constructed from S. pnuemonicae, such as the one deposited by the inventors at the ATCC for convenience, ATCC Deposit No. 97755, or from DNA isolated from the same organism such as the S. pneumoniae strain deposited with the ATCC as Deposit No. 55840.
  • a list of PCR primers which can be used for this pu ⁇ ose is provided in Table 3, below.
  • the PCR primers anneal to the nucleotide sequences encoding both the amino terminal and carboxy terminal amino acid sequences of the desired portion of the polypeptides of Table 1.
  • nucleotides containing restriction sites to facilitate cloning in the pQElO vector were added to the 5' and 3' primer sequences, respectively. Such restriction sites are listed in Table 3 for each primer.
  • the primer comprises, from the 5' end, 4 random nucleotides to prevent "breathing" during the annealing process, a restriction site (shown in Table 3), and approximately 15 nucleotides of S. pneumoniae ORF sequence (the complete sequence of each cloning primer is shown as SEQ ED NO:227 through SEQ ID NO:452).
  • the 5' and 3' primers were selected to amplify their respective nucleotide coding sequences.
  • One of ordinary skill in the art would appreciate that the point in the protein coding sequence where the 5' primer begins may be varied to amplify a DNA segment encoding any desired portion of the complete amino acid sequences described in
  • the point in the protein coding sequence where the 3' primer begins may also be varied to amplify a DNA segment encoding any desired portion of the complete amino acid sequences described in Table 1.
  • the amplified DNA fragment and the pQElO vector are digested with the appropriate restriction enzyme(s) and the digested DNAs are then ligated together.
  • the ligation mixture is transformed into competent E. coli cells using standard procedures such as those described in Sambrook et al, Molecular Cloning: a Laboratory Manual, 2nd Ed.; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. ( 1989). Transformants are identified by their ability to grow under selective pressure on LB plates. Plasmid DNA is isolated from resistant colonies and the identity of the cloned DNA confirmed by restriction analysis, PCR and DNA sequencing.
  • Clones containing the desired constructs are grown overnight ("O/N") in liquid culture under selection.
  • the O/N culture is used to inoculate a large culture, at a dilution of approximately 1:25 to 1 :250.
  • the cells are grown to an optical density at 600 nm ("OD600") of between 0.4 and 0.6.
  • Isopropyl-b-D- thiogalactopyranoside (“IPTG”) is then added to a final concentration of 1 mM to induce transcription from the lac repressor sensitive promoter, by inactivating the lacl repressor.
  • IPTG Isopropyl-b-D- thiogalactopyranoside
  • Cells subsequently are incubated further for 3 to 4 hours. Cells are then harvested by centrifugation.
  • the cells are stirred for 3-4 hours at 4 C in 6M guanidine-HCl, pH 8.
  • the cell debris is removed by centrifugation, and the supernatant containing the protein of interest is loaded onto a nickel-nitrilo-tri-acetic acid (“NiNTA”) affinity resin column (available from QIAGEN, Inc., supra).
  • NiNTA nickel-nitrilo-tri-acetic acid
  • Proteins with a 6x His tag bind to the NI-NTA resin with high affinity and can be purified in a simple one-step procedure (for details see: The QIAexpressionist, 1995, QIAGEN, Inc., supra). Briefly, the supernatant is loaded onto the column in 6
  • the purified protein is then renatured by dialyzing it against phosphate - buffered saline (PBS) or 50 mM Na-acetate, pH 6 buffer plus 200 mM NaCl.
  • PBS phosphate - buffered saline
  • the protein can be successfully refolded while immobilized on the Ni-NTA column.
  • the recommended conditions are as follows: renature using a linear 6M-1M urea gradient in 500 mM NaCl, 20% glycerol, 20 mM Tris/HCl pH7.4, containing protease inhibitors. The renaturation should be performed over a period of 1.5 hours or more. After renaturation the proteins can be eluted by the addition of 250 mM imidazole. Imidazole is removed by a final dialyzing step against PBS or 50 mM sodium acetate pH6 buffer plus 200 mM NaCl. The purified protein is stored at 4°C or frozen at -80°C.
  • the DNA sequences encoding the amino acid sequences of Table 1 may also be cloned and expressed as fusion proteins by a protocol similar to that described directly above, wherein the pET-32b(+) vector (Novagen, 601 Science Drive, Madison, Wl 53711) is preferentially used in place of pQElO.
  • Todd Hewitt (TH) broth (Difco laboratories, Detroit, MI) with 17% FCS, and horse blood agar plates are used for culturing the bacteria. Both broth and blood plates are incubated at 37°C in a 5% CO 2 atmosphere. Blood plates are incubated for 18 hr. The culture broth is regularly 10-fold serially diluted in TH broth kept at room temperature and bacterial suspensions are kept at room temperature until challenge of mice.
  • C3H7HeJ mice (The Jackson Laboratory, Bar Harbor, ME) are injected intraperitoneally (i.p.) at week 0 with 20 g of recombinant streptococcal protein, or phosphate-buffered saline (PBS), emulsified with complete Freund's adjuvant (CFA), given a similar booster immunization in incomplete Freund's adjuvant (EFA) at week 4, and challenged at week 6.
  • PBS phosphate-buffered saline
  • CFA complete Freund's adjuvant
  • ESA incomplete Freund's adjuvant
  • S. pneumoniae are diluted in TH broth from exponentially-growing cultures and mice are injected subcutaneously (s.c.) at the base of the tail with 0.1 ml of these dilutions (serial dilutions are used to find medium infectious dose).
  • Streptococci used for challenge are passaged fewer than six times in vitro.
  • blood samples are obtained from the distal part of the lateral femoral vein into heparinized capillary tubes.
  • a 25 ul blood sample is serially 10-fold diluted in TH broth, and 25 ul of diluted and undiluted blood is plated onto blood agar plates. The plates are incubated for 18 hr. and colonies are counted.
  • ELISA and immunoblot are used to quantify levels of streptococcal-specific antibodies (ELISA and immunoblot), and to evaluate the functional properties of these antibodies (growth inhibition assay).
  • the ELISA and immunoblot assays are also used to detect and quantify antibodies elicited in response to streptococcal infection that react with specific streptococcal antigens. Where antibodies to certain streptococcal antigens are elicited by infection this is taken as evidence that the streptococcal proteins in question are expressed in vivo. Absence of infection-derived antibodies (seroconversion) following streptococcal challenge is evidence that infection is prevented or suppressed.
  • the immunoblot assay is also used to ascertain whether antibodies raised against recombinant streptococcal antigens recognize a protein of similar size in extracts of whole streptococci. Where the natural protein is of similar, or identical, size in the immunoblot assay to the recombinant version of the same protein, this is taken as evidence that the recombinant protein is the product of a full-length clone of the respective gene.
  • the ELISA is used to quantify levels of antibodies reactive with streptococcus antigens elicited in response to immunization with these streptococcal antigens.
  • Wells of 96 well microtiter plates (Immunlon 4, Dynatech, Chantilly, Virginia, or equivalent) are coated with antigen by incubating 50 1 of 1 g/ml protein antigen solution in a suitable buffer, typically 0.1 M sodium carbonate buffer at pH 9.6. After decanting unbound antigen, additional binding sites are blocked by incubating 100 1 of 3% nonfat milk in wash buffer (PBS, 0.2% Tween 20, pH 7.4).
  • a suitable buffer typically 0.1 M sodium carbonate buffer at pH 9.6.
  • PBS 0.2% Tween 20, pH 7.4
  • total streptococcal protein extracts or recombinant streptococcal antigen are boiled in SDS/2-ME sample buffer before electrophoresis through 3% acrylamide stacking gels, and resolving gels of higher acrylamide concentration, typically 10-15% acrylamide monomer.
  • Gels are electro-blotted to nitrocellulose membranes and lanes are probed with dilutions of antibody to be tested for reactivity with specific streptococcal antigens, followed by the appropriate secondary antibody-enzyme (horseradish peroxidase) conjugate.
  • secondary antibody-enzyme horseradish peroxidase conjugate.
  • membranes are stained with Ponceau S . Immunoblot signals from bound antibodies are detected on x-ray film as chemiluminescence using ECLTM reagents (Amersham Co ⁇ ., Arlington Heights, Illinois).
  • Northern blot analysis is carried out using methods described by, among others, Sambrook et al, supra, to detect the expression of the S. pneumoniae nucleotide sequences of the present invention in animal tissues.
  • a cDNA probe containing an entire nucleotide sequence shown in Table 1 is labeled with 32p using the redip ⁇ meTM DNA labeling system (Amersham Life Science), according to manufacturer's instructions. After labeling, the probe is purified using a CHROMA SPIN- 100TM column (Clontech Laboratories, Inc.), according to manufacturer's protocol number PT 1200- 1. The purified labeled probe is then used to detect the expression of Streptococcus mRNA in an animal tissue sample.
  • LGVTHIQLLPVLSYYFVNELKNHERLSDYASSNSNYN GYDPQNYFSLTGMYSSDPKNPEKRIAEFKNL INEIHKRGMGAILDWYNHTAKV ⁇ LFEDLEPNYYHF DADGTPRTSFGGGRLGTTHHMTKRLLIDSIKY LVDTYKVDGFRFDMMGDHDAAS I EEAYKAARA NPNLI LGEGWRTYAGDENMPTKAADQD MKHTDTV Table 1 64
  • SP052 nucleotide SEQ ID NO: 83
  • SP054 nucleotide SEQ ID NO: 87
  • FNPTVGTF FTAGLSL VLLVSKRENGKKRLVHFLLLTSMGVQLLPA ⁇ AFGLTSQILSAYNSQLSIGVG EH PEPLKIEGYQYIGYIKTKKQDNTELSRTV ⁇ GKYSAQRDSQPNSTKTSDWHSADLEWNQGQGKVSL QGEA ⁇ GDDGLSEKSSIAADNLSSNDSFASQVEQNPDHKGESWRPTVPEQGNPVSATTVQSAEEEVLAT TNDRPEYK PLETKGTQEPGHEGEAAVREDLPVYTKPLETKGTQGPGHEGEAAVREEEPAYTEPLATKG TQEPGHEGKATVREETLEYTEPVATKGTQEPEHEGERXVEEELPALEVTTR RTEIQNIPYTTEEIQDP TLLKNRRKIERQGQAGTRTIQYEDYIVNGLWVETKEVSRTEVAPVNE ⁇ /VKVGTLVKVKPTVEITNLTKV ENKKSITVSYNLIDTTS
  • Glu-1 to Lys-13 Pro-24 to Gly-36; Val-104 to Thr-112; Ala-118 to Asn- 130; Trp-137 to Ala-146; Ser-151 to Ile-159; Ile-181 to Leu-188; and Pro-194 to Tyr-202.
  • Gly-1 to Asn-7 Tyr-24 to Gln-34; His-47 to Phe-55; Ser-60 to Ala-67; Ala-122 to Leu-129; Leu-221 to Lys-230; Val-236 to Phe-256; and Asp-271 to Gly-283; and Leu-291 to Asp-297.
  • Leu-4 to Lys-17 Gln-24 to Leu-32; Asp-60 to Ser-66; Ser-70 to Asp-76; Ala-276 to Lys-283; Asn-304 to Lys-311; and Thr-429 to Pro-437.
  • Ser-1 to Thr-12 Ala-36 to Tyr-45; Gln-48 to Ile-54; Lys-59 to Lys-76; Tyr-113 to Leu-138; and Phe-212 to Asp-219.
  • Asp-8 to Val-16 Gly-27 to Thr-35; Gly-178 to Asp-195; Thr-200 to Asp209; Trp-218 to Leu-224; and Lys-226 to Asp-241.
  • Tyr-4 to Asp-25; Lys-33 to Val-70; Asp-151 to Thr-170; Asp-222 to Val-257; Thr-290 to Phe-301; and Gly-357 to Val-367.
  • Lys-6 to Tyr-16 Gln-19 to Ile-27; Arg-50 to Ala-58; Leu-112 to Val- 128; Ile-151 to Asn-167; Leu-305 to Phe-321.
  • Glu-37 to Val-45 Glu-55 to Val-68; Thr-104 to Thr-119; Ile-127 to Tyr-135; Asn-220 to Ile-232; Thr-237 to Ala-250; Ser-253 to Ala-263; Glu-284 to Ile-297; and Met-438 to Asn-455.
  • Gly-184 Tyr-194 to Ala-202; Ala-217 to Gly-225; and Asn-85 to Gly-93.
  • Lys-422 Pro-421 to Asn-429; Trp-465 to Tyr-473; Phe-487 to Tyr-495;
  • Gln-175 Pro-189 to Glu-197; Gly-207 to Leu-215; Ser-219 to Gln-227;

Abstract

The present invention relates to novel vaccines for the prevention or attenuation of infection by Streptococcus pneumoniae. The invention further relates to isolated nucleic acid molecules encoding antigenic polypeptides of Streptococcus pneumoniae. Antigenic polypeptides are also provided, as are vectors, host cells and recombinant methods for producing the same. The invention additionally relates to diagnostic methods for detecting Streptococcus nucleic acids, polypeptides and antibodies in a biological sample.

Description

Streptococcus pneumoniae Antigens and Vaccines
Field of the Invention
The present invention relates to novel Streptococcus pneumoniae antigens for the detection of Streptococcus and for the prevention or attenuation of disease caused by Streptococcus. The invention further relates to isolated nucleic acid molecules encoding antigenic polypeptides of S. pneumoniae. Antigenic polypeptides are also provided, as are vectors, host cells and recombinant methods for producing the same. The invention additionally relates to diagnostic methods for detecting Streptococcus gene expression.
Background of the Invention
Streptococcus pneumoniae has been one of the most extensively studied microorganisms since its first isolation in 1881. It was the object of many investigations that led to important scientific discoveries. In 1928, Griffith observed that when heat-killed encapsulated pneumococci and live strains constitutively lacking any capsule were concomitantly injected into mice, the nonencapsulated could be converted into encapsulated pneumococci with the same capsular type as the heat-killed strain. Years later, the nature of this "transforming principle," or carrier of genetic information, was shown to be
DNA. (Avery, O.T., et al, J. Exp. Med., 79: 137- 157 ( 1944)).
In spite of the vast number of publications on S. pneumoniae many questions about its virulence are still unanswered, and this pathogen remains a major causative agent of serious human disease, especially community-acquired pneumonia. (Johnston, R.B., et al., Rev. Infect. Dis. 73(Suppl. 6):S509-517
(1991)). In addition, in developing countries, the pneumococcus is responsible for the death of a large number of children under the age of 5 years from pneumococcal pneumonia. The incidence of pneumococcal disease is highest in infants under 2 years of age and in people over 60 years of age. Pneumococci are the second most frequent cause (after Haemophilus influenzae type b) of bacterial meningitis and otitis media in children. With the recent introduction of conjugate vaccines for H. influenzae type b, pneumococcal meningitis is likely to become increasingly prominent. S. pneumoniae is the most important etiologic agent of community-acquired pneumonia in adults and is the second most common cause of bacterial meningitis behind Neisseria meningitidis.
The antibiotic generally prescribed to treat S. pneumoniae is benzylpenicillin, although resistance to this and to other antibiotics is found occasionally. Pneumococcal resistance to penicillin results from mutations in its penicillin-binding proteins. In uncomplicated pneumococcal pneumonia caused by a sensitive strain, treatment with penicillin is usually successful unless started too late. Erythromycin or clindamycin can be used to treat pneumonia in patients hypersensitive to penicillin, but resistant strains to these drugs exist. Broad spectrum antibiotics (e.g., the tetracyclines) may also be effective, although tetracycline-resistant strains are not rare. In spite of the availability of antibiotics, the mortality of pneumococcal bacteremia in the last four decades has remained stable between 25 and 29%. (Gillespie, S.H., et al., J. Med. Microbiol. 2S/237-248 ( 1989). S. pneumoniae is carried in the upper respiratory tract by many healthy individuals. It has been suggested that attachment of pneumococci is mediated by a disaccharide receptor on fibronectin, present on human pharyngeal epithelial cells. (Anderson, B.J., et al, J. Immunol. 742:2464-2468 ( 1989). The mechanisms by which pneumococci translocate from the nasopharynx to the lung, thereby causing pneumonia, or migrate to the blood, giving rise to bacteremia or septicemia, are poorly understood. (Johnston, R.B ., et al., Rev. Infect. Dis. 73(Suppl. 6):S509-517 (1991).
Various proteins have been suggested to be involved in the pathogenicity of S. pneumoniae, however, only a few of them have actually been confirmed as virulence factors. Pneumococci produce an IgAl protease that might interfere with host defense at mucosal surfaces. (Kornfield, S.J., et al. , Rev. Inf. Dis. 5:521-534 ( 1981 ). S. pneumoniae also produces neuraminidase, an enzyme that may facilitate attachment to epithelial cells by cleaving sialic acid from the host glycolipids and gangliosides. Partially purified neuraminidase was observed to induce meningitis-like symptoms in mice; however, the reliability of this finding has been questioned because the neuraminidase preparations used were probably contaminated with cell wall products. Other pneumococcal proteins besides neuraminidase are involved in the adhesion of pneumococci to epithelial and endothelial cells. These pneumococcal proteins have as yet not been identified. Recently, Cundell et al. , reported that peptide permeases can modulate pneumococcal adherence to epithelial and endothelial cells. It was, however, unclear whether these permeases function directly as adhesions or whether they enhance adherence by modulating the expression of pneumococcal adhesions. (DeVelasco, E.A., et al., Micro. Rev. 59:591-603 ( 1995). A better understanding of the virulence factors determining its pathogenicity will need to be developed to cope with the devastating effects of pneumococcal disease in humans. Ironically, despite the prominent role of S. pneumoniae in the discovery of DNA, little is known about the molecular genetics of the organism. The S . pneumoniae genome consists of one circular, covalently closed, double- stranded DNA and a collection of so-called variable accessory elements, such as prophages, plasmids, transposons and the like. Most physical characteristics and almost all of the genes of S. pneumoniae are unknown. Among the few that have been identified, most have not been physically mapped or characterized in detail. Only a few genes of this organism have been sequenced. (See, for instance current versions of GENBANK and other nucleic acid databases, and references that relate to the genome of S. pneumoniae such as those set out elsewhere herein.) Identification of in vø-expressed, and broadly protective, antigens of 5. pneumoniae has remained elusive.
Summary of the Invention The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding the S. pneumoniae polypeptides described in Table 1 and having the amino acid sequences shown as SEQ LD NO:2, SEQ ID NO:4, SEQ ID NO:6, and so on through SEQ ID NO:226. Thus, one aspect of the invention provides isolated nucleic acid molecules comprising polynucleotides having a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence encoding any of the amino acid sequences of the polypeptides shown in Table 1 ; and (b) a nucleotide sequence complementary to any of the nucleotide sequences in (a).
Further embodiments of the invention include isolated nucleic acid molecules that comprise a polynucleotide having a nucleotide sequence at least
90% identical, and more preferably at least 95%, 96%, 97%, 98% or 99% identical, to any of the nucleotide sequences in (a) or (b) above, or a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide in (a) or (b) above. This polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues. Additional nucleic acid embodiments of the invention relate to isolated nucleic acid molecules comprising polynucleotides which encode the amino acid sequences of epitope-bearing portions of an S. pneumoniae polypeptide having an amino acid sequence in (a) above.
The present invention also relates to recombinant vectors, which include the isolated nucleic acid molecules of the present invention, and to host cells containing the recombinant vectors, as well as to methods of making such vectors and host cells and for using these vectors for the production of S. pneumoniae polypeptides or peptides by recombinant techniques.
The invention further provides isolated S. pneumoniae polypeptides having an amino acid sequence selected from the group consisting of an amino acid sequence of any of the polypeptides described in Table 1.
The polypeptides of the present invention also include polypeptides having an amino acid sequence with at least 70% similarity, and more preferably at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% similarity to those described in Table 1 , as well as polypeptides having an amino acid sequence at least 70% identical, more preferably at least 75% identical, and still more preferably 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to those above; as well as isolated nucleic acid molecules encoding such polypeptides.
The present invention further provides a vaccine, preferably a multi-component vaccine comprising one or more of the S. pneumoniae polynucleotides or polypeptides described in Table 1 , or fragments thereof, together with a pharmaceutically acceptable diluent, carrier, or excipient, wherein the S. pneumoniae polypeptide(s) are present in an amount effective to elicit an immune response to members of the Streptococcus genus in an animal. The S. pneumoniae polypeptides of the present invention may further be combined with one or more immunogens of one or more other streptococcal or non-streptococcal organisms to produce a multi-component vaccine intended to elicit an immunological response against members of the Streptococcus genus and, optionally, one or more non-streptococcal organisms. The vaccines of the present invention can be administered in a DNA form, e.g., "naked" DNA, wherein the DNA encodes one or more streptococcal polypeptides and, optionally, one or more polypeptides of a non-streptococcal organism. The DNA encoding one or more polypeptides may be constructed such that these polypeptides are expressed fusion proteins. The vaccines of the present invention may also be administered as a component of a genetically engineered organism. Thus, a genetically engineered organism which expresses one or more S. pneumoniae polypeptides may be administered to an animal. For example, such a genetically engineered organism may contain one or more S. pneumoniae polypeptides of the present invention intracellularly, on its cell surface, or in its periplasmic space. Further, such a genetically engineered organism may secrete one or more S. pneumoniae polypeptides. The vaccines of the present invention may be co-administered to an animal with an immune system modulator (e.g., CD86 and GM-CSF).
The invention also provides a method of inducing an immunological response in an animal to one or more members of the Streptococcus genus, preferrably one or more isolates of the S. pneumoniae genus, comprising administering to the animal a vaccine as described above.
The invention further provides a method of inducing a protective immune response in an animal, sufficient to prevent or attenuate an infection by members of the Streptococcus genus, preferrably at least S. pneumoniae, comprising administering to the animal a composition comprising one or more of the polynucleotides or polypeptides described in Table 1 , or fragments thereof. Further, these polypeptides, or fragments thereof, may be conjugated to another immunogen and/or administered in admixture with an adjuvant.
The invention further relates to antibodies elicited in an animal by the administration of one or more S. pneumoniae polypeptides of the present invention and t methods for producing such antibodies.
The invention also provides diagnostic methods for detecting the expression of genes of members of the Streptococcus genus in an animal. One such method involves assaying for the expression of a gene encoding S. pneumoniae peptides in a sample from an animal. This expression may be assayed either directly (e.g. , by assaying polypeptide levels using antibodies elicited in response to amino acid sequences described in Table 1) or indirectly (e.g. , by assaying for antibodies having specificity for amino acid sequences described in Table 1). An example of such a method involves the use of the polymerase chain reaction (PCR) to amplify and detect Streptococcus nucleic acid sequences.
The present invention also relates to nucleic acid probes having all or part of a nucleotide sequence described in Table 1 (shown as SEQ ID NO: l, SEQ ID NO:3, SEQ ID NO:5, and so on through SEQ ID NO:225) which are capable of hybridizing under stringent conditions to Streptococcus nucleic acids.
The invention further relates to a method of detecting one or more Streptococcus nucleic acids in a biological sample obtained from an animal, said one or more nucleic acids encoding Streptococcus polypeptides, comprising: (a) contacting the sample with one or more of the above-described nucleic acid probes, under conditions such that hybridization occurs, and (b) detecting hybridization of said one or more probes to the Streptococcus nucleic acid present in the biological sample. The invention also includes immunoassays, including an immunoassay for detecting Streptococcus, preferrably at least isolates of the S. pneumoniae genus, comprising incubation of a sample (which is suspected of being infected with Streptococcus) with a probe antibody directed against an antigen/epitope of S. pneumoniae, to be detected under conditions allowing the formation of an antigen-antibody complex; and detecting the antigen-antibody complex which contains the probe antibody. An immunoassay for the detection of antibodies which are directed against a Streptococcus antigen comprising the incubation of a sample (containing antibodies from a mammal suspected of being infected with Streptococcus) with a probe polypeptide including an epitope of S. pneumoniae , under conditions that allow the formation of antigen-antibody complexes which contain the probe epitope containing antigen.
Some aspects of the invention pertaining to kits are those for: investigating samples for the presence of polynucleotides derived from Streptococcus which comprise a polynucleotide probe including a nucleotide sequence selected from Table 1 or a fragment thereof of approximately 15 or more nucleotides, in an appropriate container; analyzing the samples for the presence of antibodies directed against a Streptococcus antigen made up of a polypeptide which contains a S. pneumoniae epitope present in the polypeptide, in a suitable container; and analyzing samples for the presence of Streptococcus antigens made up of an anti-5. pneumoniae antibody, in a suitable container.
Detailed Description
The present invention relates to recombinant antigenic S. pneumoniae polypeptides and fragments thereof. The invention also relates to methods for using these polypeptides to produce immunological responses and to confer immunological protection to disease caused by members of the genus Streptococcus, at least isolates of the S. pneumoniae genus. The invention further relates to nucleic acid sequences which encode antigenic S. pneumoniae polypeptides and to methods for detecting S. pneumoniae nucleic acids and polypeptides in biological samples. The invention also relates to S. pneumoniae -specific antibodies and methods for detecting such antibodies produced in a host animal.
Definitions
The following definitions are provided to clarify the subject matter which the inventors consider to be the present invention. As used herein, the phrase "pathogenic agent" means an agent which causes a disease state or affliction in an animal. Included within this definition, for examples, are bacteria, protozoans, fungi, viruses and metazoan parasites which either produce a disease state or render an animal infected with such an organism susceptible to a disease state (e.g., a secondary infection). Further included are species and strains of the genus Streptococcus which produce disease states in animals.
As used herein, the term "organism" means any living biological system, including viruses, regardless of whether it is a pathogenic agent. As used herein, the term "Streptococcus" means any species or strain of bacteria which is members of the genus Streptococcus. Such species and strains are known to those of skill in the art, and include those that are pathogenic and those that are not.
As used herein, the phrase "one or more S. pneumoniae polypeptides of the present invention" means polypeptides comprising the amino acid sequence of one or more, of the S. pneumoniae polypeptides described in Table 1 and disclosed as SEQ ID NO:2, SEQ ID NO:4, SEQ ED NO:6, and so on through SEQ ID NO:226. These polypeptides may be expressed as fusion proteins wherein the 5. pneumoniae polypeptides of the present invention are linked to additional amino acid sequences which may be of streptococcal or non- streptococcal origin. This phrase further includes polypeptide comprising fragments of the S. pneumoniae polypeptides of the present invention.
Additional definitions are provided throughout the specification.
Explanation of Table 1
Table 1, below, provides information describing 1 13 open reading frames (ORFs) which encode potentially antigenic polypeptides of S. pneumoniae of the present invention. The table lists the ORF identifier which consists of the letters SP, which denote S. pneumoniae, followed immediately by a three digit numeric code, which arbitrarily number the potentially antigenic polypeptides of S. pneumoniae of the present invention and the nucleotide or amino acid sequence of each ORF and encoded polypeptide. The table further correlates the ORF identifier with a sequence identification number (SEQ ID NO:). The actual nucleotide or amino acid sequence of each ORF identifier is also shown in the Sequence Listing under the corresponding SEQ ID NO.
Thus, for example, the designation "SPI 26" refers to both the nucleotide and amino acid sequences of S. pneumoniae polypeptide number 126 of the present invention. Further, "SP126" correlates with the nucleotide sequence shown as SEQ ED NO:223 and with the amino acid sequence shown as SEQ ID NO:224 as is described in Table 1.
The open reading frame within each "ORF" begins with the second nucleotide shown. Thus, the first codon for each nucleotide sequence shown is bases 2-4, the second 5-7, the third 8-10, and so on.
Explanation of Table 2
Table 2 lists the antigenic epitopes present in each of the S. pneumoniae polypeptides described in Table 1 as predicted by the inventors. Each S . pneumoniae polypeptide shown in Table 1 has one or more antigenic epitopes described in Table 2. It will be appreciated that depending on the analytical criteria used to predict antigenic determinants, the exact address of the determinant may vary slightly. The exact location of the antigenic determinant may shift by about 1 to 5 residues, more likely 1 to 2 residues, depending on the criteria used. Thus, the first antigenic determinant described in Table 2 ,
"Lys-1 to Ile-10" of SP001 , represents a peptide comprising the lysine at position 1 in SEQ ED NO:2 through and including the isoleucine at position 10 in SEQ ID NO:2, but may include more or fewer residues than those 10. It will also be appreciated that, generally speaking, amino acids can be added to either terminus of a peptide or polypeptide containing an antigenic epitope without affecting its activity, whereas removing residues from a peptide or polypeptide containing only the antigenic determinant is much more likely to destroy activity. It will be appreciated that the residues and locations shown described in Table 2 correspond to the amino acid sequences for each ORF shown in Table 1 and in the Sequence Listing.
Explanation of Table 3
Table 3 shows PCR primers designed by the inventors for the amplification of polynucleotides encoding polypeptides of the present invention according to the method of Example 1. PCR primer design is routine in the art and those shown in Table 3 are provided merely for the convenience of the skilled artisan. It will be appreciated that others can be used with equal success.
For each primer, the table lists the corresponding ORF designation from
Table 1 followed by either an "A" or a "B". The "A" primers are the 5' primers and the "B" primers 3'. A restriction enzyme site was built into each primer to allow ease of cloning. The restriction enzyme which will recognize and cleave a sequence within each primer is shown in Table 3, as well, under the heading "RE" for restriction enzyme. Finally the sequence identifier is shown in Table 3 for each primer for easy correlation with the Sequence Listing.
Selection of Nucleic Acid Sequences Encoding Antigenic S . pneumoniae Polypeptides
The present invention provides a select number of ORFs from those presented in the fragments of the S. pneumoniae genome which may prove useful for the generation of a protective immune response. The sequenced S . pneumoniae genomic DNA was obtained from a sub-cultured isolate of S . pneumoniae Strain 7/87 14.8.91 , which has been deposited at the American
Type Culture Collection, as a convenience to those of skill in the art. The 5. pneumoniae isolate was deposited on October 10, 1996 at the ATCC, 12301 Park Lawn Drive, Rockville, Maryland 20852, and given accession number 55840. A genomic library constructed from DNA isolated from the S. pneumoniae isolate was also deposited at the ATCC on October 1 1 , 1996 and given ATCC E^eposit No. 97755. A more complete listing of the sequence obtained from the S. pneumoniae genome may be found in co-pending U.S . Provisional Application Serial No. 60/029,960, filed 10/31/96, incoφorated herein by reference in its entirety. Some ORFs contained in the subset of fragments of the S. pneumoniae genome disclosed herein were derived through the use of a number of screening criteria detailed below.
The selected ORFs do not consist of complete ORFs. Although a polypeptide representing a complete ORF may be the closest approximation of a protein native to an organism, it is not always preferred to express a complete ORF in a heterologous system. It may be challenging to express and purify a highly hydrophobic protein by common laboratory methods. Thus, the polypeptide vaccine candidates described herein may have been modified slightly to simplify the production of recombinant protein. For example, nucleotide sequences which encode highly hydrophobic domains, such as those found at the amino terminal signal sequence, have been excluded from some constructs used for in vitro expression of the polypeptides. Furthermore, any highly hydrophobic amino acid sequences occurring at the carboxy terminus have also been excluded from the recombinant expression constructs. Thus, in one embodiment, a polypeptide which represents a truncated or modified ORF may be used as an antigen.
While numerous methods are known in the art for selecting potentially immunogenic polypeptides, many of the ORFs disclosed herein were selected on the basis of screening all theoretical 5. pneumoniae ORFs for several aspects of potential immunogenicity. One set of selection criteria are as follows:
1. Type I signal sequence: An amino terminal type I signal sequence generally directs a nascent protein across the plasma and outer membranes to the exterior of the bacterial cell. Experimental evidence obtained from studies with
Escherichia coli suggests that the typical type I signal sequence consists of the following biochemical and physical attributes (Izard, J. W. and Kendall, D. A. Mol. Microbiol. 13:765-773 (1994)). The length of the type I signal sequence is approximately 15 to 25 primarily hydrophobic amino acid residues with a net positive charge in the extreme amino terminus. In addition, the central region of the signal sequence adopts an alpha-helical conformation in a hydrophobic environment. Finally, the region surrounding the actual site of cleavage is ideally six residues long, with small side-chain amino acids in the -1 and -3 positions. 2. Type IV signal sequence: The type IV signal sequence is an example of the several types of functional signal sequences which exist in addition to the type I signal sequence detailed above. Although functionally related, the type IV signal sequence possesses a unique set of biochemical and physical attributes (Strom, M. S. and Lory, S., J. Bacteriol. 174:7345-735 1 ( 1992)). These are typically six to eight amino acids with a net basic charge followed by an additional sixteen to thirty primarily hydrophobic residues. The cleavage site of a type IV signal sequence is typically after the initial six to eight amino acids at the extreme amino terminus. In addition, type EV signal sequences generally contain a phenylalanine residue at the +1 site relative to the cleavage site. 3. Lipoprotein: Studies of the cleavage sites of twenty-six bacterial lipoprotein precursors has allowed the definition of a consensus amino acid sequence for lipoprotein cleavage. Nearly three-fourths of the bacterial lipoprotein precursors examined contained the sequence L-(A,S)-(G,A)-C at positions -3 to +1, relative to the point of cleavage (Hayashi, S. and Wu, H . C, J. Bioenerg. Biomembr. 22:451-471 ( 1990)).
4. LPXTG motif: It has been experimentally determined that most anchored proteins found on the surface of gram-positive bacteria possess a highly conserved carboxy terminal sequence. More than fifty such proteins from organisms such as S. pyogenes, S. mutans, E. faecalis, S. pneumoniae, and others, have been identified based on their extracellular location and carboxy terminal amino acid sequence (Fischetti, V. A., ASM News 62:405-410 ( 1996)). The conserved region consists of six charged amino acids at the extreme carboxy terminus coupled to 15-20 hydrophobic amino acids presumed to function as a transmembrane domain. Immediately adjacent to the transmembrane domain is a six amino acid sequence conserved in nearly all proteins examined. The amino acid sequence of this region is L-P-X-T-G-X, where X is any amino acid. An algorithm for selecting antigenic and immunogenic S. pneumoniae polypeptides including the foregoing criteria was developed. Use of the algorithm by the inventors to select immunologically useful S. pneumoniae polypeptides resulted in the selection of a number of the disclosed ORFs. Polypeptides comprising the polypeptides identified in this group may be produced by techniques standard in the art and as further described herein.
Nucleic Acid Molecules
The present invention provides isolated nucleic acid molecules comprising polynucleotides encoding the S. pneumoniae polypeptides having the amino acid sequences described in Table 1 and shown as SEQ ED NO:2,
SEQ ID NO:4, SEQ ID NO:6, and so on through SEQ ED NO:226, which were determined by sequencing the genome of S. pneumoniae and selected as putative immunogens.
Unless otherwise indicated, all nucleotide sequences determined by sequencing a DNA molecule herein were determined using an automated DNA sequencer (such as the Model 373 from Applied Biosystems, Inc.), and all amino acid sequences of polypeptides encoded by DNA molecules determined herein were predicted by translation of DNA sequences determined as above. Therefore, as is known in the art for any DNA sequence determined by this automated approach, any nucleotide sequence determined herein may contain some errors. Nucleotide sequences determined by automation are typically at least about 90% identical, more typically at least about 95% to at least about 99.9% identical to the actual nucleotide sequence of the sequenced DNA molecule. The actual sequence can be more precisely determined by other approaches including manual DNA sequencing methods well known in the art.
As is also known in the art, a single insertion or deletion in a determined nucleotide sequence compared to the actual sequence will cause a frame shift in translation of the nucleotide sequence such that the predicted amino acid sequence encoded by a determined nucleotide sequence will be completely different from the amino acid sequence actually encoded by the sequenced DNA molecule, beginning at the point of such an insertion or deletion.
Unless otherwise indicated, each "nucleotide sequence" set forth herein is presented as a sequence of deoxyribonucleotides (abbreviated A, G , C and T). However, by "nucleotide sequence" of a nucleic acid molecule or polynucleotide is intended, for a DNA molecule or polynucleotide, a sequence of deoxyribonucleotides, and for an RNA molecule or polynucleotide, the corresponding sequence of ribonucleotides (A, G, C and U), where each thymidine deoxyribonucleotide (T) in the specified deoxyribonucleotide sequence is replaced by the ribonucleotide uridine (U). For instance, reference to an RNA molecule having a sequence described in Table 1 set forth using deoxyribonucleotide abbreviations is intended to indicate an RNA molecule having a sequence in which each deoxyribonucleotide A, G or C described in Table 1 has been replaced by the corresponding ribonucleotide A, G or C, and each deoxyribonucleotide T has been replaced by a ribonucleotide U.
Nucleic acid molecules of the present invention may be in the form of RNA, such as mRNA, or in the form of DNA, including, for instance, cDNA and genomic DNA obtained by cloning or produced synthetically. The DNA may be double-stranded or single-stranded. Single-stranded DNA or RNA may be the coding /Strand, also known as the sense strand, or it may be the non-coding strand, also referred to as the anti-sense strand.
By "isolated" nucleic acid molecule(s) is intended a nucleic acid molecule, DNA or RNA, which has been removed from its native environment. For example, recombinant DNA molecules contained in a vector are considered isolated for the purposes of the present invention. Further examples of isolated DNA molecules include recombinant DNA molecules maintained in heterologous host cells or purified (partially or substantially) DNA molecules in solution. Isolated RNA molecules include in vivo or in.vitro RNA transcripts of the DNA molecules of the present invention. Isolated nucleic acid molecules according to the present invention further include such molecules produced synthetically.
Isolated nucleic acid molecules of the present invention include DNA molecules comprising a nucleotide sequence described in Table 1 and shown as SEQ ED NO: l , SEQ ID NO:3, SEQ ID NO:5, and so on through SEQ ED
NO:225; DNA molecules comprising the coding sequences for the polypeptides described in Table 1 and shown as SEQ ED NO:2, SEQ ED NO:4, SEQ ED NO:6, and so on through SEQ ID NO:226; and DNA molecules which comprise sequences substantially different from those described above but which, due to the degeneracy of the genetic code, still encode the S. pneumoniae polypeptides described in Table 1. Of course, the genetic code is well known in the art. Thus, it would be routine for one skilled in the art to generate such degenerate variants. The invention also provides nucleic acid molecules having sequences complementary to any one of those described in Table 1. Such isolated molecules, particularly DNA molecules, are useful as probes for detecting expression of Streptococcal genes, for instance, by Northern blot analysis or the polymerase chain reaction (PCR).
The present invention is further directed to fragments of the isolated nucleic acid molecules described herein. By a fragment of an isolated nucleic acid molecule having a nucleotide sequence described in Table 1, is intended fragments at least about 15 nt, and more preferably at least about 17 nt, still more preferably at least about 20 nt, and even more preferably, at least about 25 nt in length which are useful as diagnostic probes and primers as discussed herein. Of course, larger fragments 50-100 nt in length are also useful according to the present invention as are fragments corresponding to most, if not all, of a nucleotide sequence described in Table 1. By a fragment at least 20 nt in length, for example, is intended fragments which include 20 or more contiguous bases of a nucleotide sequence as described in Table 1. Since the nucleotide sequences identified in Table 1 are provided as SEQ ED NO: l , SEQ ID NO:3, SEQ ID NO:5, and so on through SEQ ED NO:225, generating such DNA fragments would be routine to the skilled artisan. For example, such fragments could be generated synthetically.
Preferred nucleic acid fragments of the present invention also include nucleic acid molecules comprising nucleotide sequences encoding epitope-bearing portions of the S. pneumoniae polypeptides identified in Table 1. Such nucleic acid fragments of the present invention include, for example, nucleotide sequences encoding polypeptide fragments comprising from about the amino terminal residue to about the carboxy terminal residue of each fragment shown in Table 2. The above referred to polypeptide fragments are antigenic regions of the S. pneumoniae polypeptides identified in Table 1.
In another aspect, the invention provides isolated nucleic acid molecules comprising polynucleotides which hybridize under stringent hybridization conditions to a portion of a polynucleotide in a nucleic acid molecule of the invention described above, for instance, a nucleic acid sequence identified in Table 1. By "stringent hybridization conditions" is intended overnight incubation at 42°C in a solution comprising: 50% formamide, 5x SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5x
Denhardt's solution, 10% dextran sulfate, and 20 g/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0. lx SSC at about 65°C. By polynucleotides which hybridize to a "portion" of a polynucleotide is intended polynucleotides (either DNA or RNA) which hybridize to at least about 15 nucleotides (nt), and more preferably at least about 17 nt, still more preferably at least about 20 nt, and even more preferably about 25-70 nt of the reference polynucleotide. These are useful as diagnostic probes and primers as discussed above and in more detail below.
Of course, polynucleotides hybridizing to a larger portion of the reference polynucleotide, for instance, a portion 50-100 nt in length, or even to the entire length of the reference polynucleotide, are also useful as probes according to the present invention, as are polynucleotides corresponding to most, if not all, of a nucleotide sequence as identified in Table 1. By a portion of a polynucleotide of "at least 20 nt in length," for example, is intended 20 or more contiguous nucleotides from the nucleotide sequence of the reference polynucleotide (e.g., a nucleotide sequences as described in Table 1). As noted above, such portions are useful diagnostically either as probes according to conventional DNA hybridization techniques or as primers for amplification of a target sequence by PCR, as described in the literature (for instance, in Molecular Cloning, A Laboratory Manual, 2nd. edition, Sambrook, J., Fritsch, E. F. and Maniatis, T., eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. ( 1989), the entire disclosure of which is hereby incoφorated herein by reference).
Since nucleic acid sequences encoding the S. pneumoniae polypeptides of the present invention are identified in Table 1 and provided as SEQ ED NO: 1 , SEQ ID NO:3, SEQ ED NO:5, and so on through SEQ ED NO:225, generating polynucleotides which hybridize to portions of these sequences would be routine to the skilled artisan. For example, the hybridizing polynucleotides of the present invention could be generated synthetically according to known techniques.
As indicated, nucleic acid molecules of the present invention which encode S. pneumoniae polypeptides of the present invention may include, but are not limited to those encoding the amino acid sequences of the polypeptides by themselves; and additional coding sequences which code for additional amino acids, such as those which provide additional functionalities. Thus, the sequences encoding these polypeptides may be fused to a marker sequence, such as a sequence encoding a peptide which facilitates purification of the fused polypeptide. In certain preferred embodiments of this aspect of the invention, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (Qiagen, Inc.), among others, many of which are commercially available. As described by Gentz and colleagues (Proc. Natl. Acad. Sci. USA 86:821-824 ( 1989)), for instance, hexa-histidine provides for convenient purification of the resulting fusion protein.
Thus, the present invention also includes genetic fusions wherein the S . pneumoniae nucleic acid sequences coding sequences identified in Table 1 are linked to additional nucleic acid sequences to produce fusion proteins. These fusion proteins may include epitopes of streptococcal or non-streptococcal origin designed to produce proteins having enhanced immunogenicity. Further, the fusion proteins of the present invention may contain antigenic determinants known to provide helper T-cell stimulation, peptides encoding sites for post-translational modifications which enhance immunogenicity (e.g. , acylation), peptides which facilitate purification (e.g., histidine "tag"), or amino acid sequences which target the fusion protein to a desired location (e.g. , a heterologous leader sequence). In all cases of bacterial expression, an N-terminal methionine residues is added. In many cases, however, the N-terminal methionine residues is cleaved off post-translationally. Thus, the invention includes polypeptides shown in Table 1 with, and without an N-termainal methionine.
The present invention thus includes nucleic acid molecules and sequences which encode fusion proteins comprising one or more S. pneumoniae polypeptides of the present invention fused to an amino acid sequence which allows for post-translational modification to enhance immunogenicity. This post-translational modification may occur either in vitro or when the fusion protein is expressed in vivo in a host cell. An example of such a modification is the introduction of an amino acid sequence which results in the attachment of a lipid moiety.
Thus, as indicated above, the present invention includes genetic fusions wherein a S. pneumoniae nucleic acid sequence identified in Table 1 is linked to a nucleotide sequence encoding another amino acid sequence. These other amino acid sequences may be of streptococcal origin (e.g. , another sequence selected from Table 1) or non-streptococcal origin.
The present invention further relates to variants of the nucleic acid molecules of the present invention, which encode portions, analogs or derivatives of the S. pneumoniae polypeptides described in Table 1. Variants may occur naturally, such as a natural allelic variant. By an "allelic variant" is intended one of several alternate forms of a gene occupying a given locus on a chromosome of an organism (Genes II, Lewin, B., ed., John Wiley & Sons, New York ( 1985)). Non-naturally occurring variants may be produced using art-known mutagenesis techniques.
Such variants include those produced by nucleotide substitutions, deletions or additions. The substitutions, deletions or additions may involve one or more nucleotides. These variants may be altered in coding regions, non-coding regions, or both. Alterations in the coding regions may produce conservative or non-conservative amino acid substitutions, deletions or additions. Especially preferred among these are silent substitutions, additions and deletions, which do not alter the properties and activities of the S. pneumoniae polypeptides disclosed herein or portions thereof. Silent substitution are most likely to be made in non-epitopic regions. Guidance regarding those regions containing epitopes is provided herein, for example, in
Table 2. Also especially preferred in this regard are conservative substitutions.
Further embodiments of the invention include isolated nucleic acid molecules comprising a polynucleotide having a nucleotide sequence at least
90% identical and more preferably at least 95%, 96%, 97%, 98% or 99% identical to: (a) a nucleotide sequence encoding any of the amino acid sequences of the polypeptides identified in Table 1; and (b) a nucleotide sequence complementary to any of the nucleotide sequences in (a) above. By a polynucleotide having a nucleotide sequence at least, for example,
95% "identical" to a reference nucleotide sequence encoding a S. pneumoniae polypeptide described in Table 1 , is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding the subject S . pneumoniae polypeptide. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence.
Certain nucleotides within some of the nucleic acid sequences shown in Table 1 were ambiguous upon sequencing. Completely unknown sequences are shown as an "N". Other unresolved nucleotides are known to be either a purine, shown as "R", or a pyrimidine, shown as "Y". Accordingly, when determining identity between two nucleotide sequences, identity is met where any nucleotide, including an "R", "Y" or "N", is found in a test sequence and at the corresponding position in the referece sequence (from Table 1). Likewise, an A, G or "R" in a test sequence is identical to an "R" in the reference sequence; and a T, C or "Y" in a test sequence is identical to a "Y" in the reference sequence.
As a practical matter, whether any particular nucleic acid molecule is at least 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, a nucleotide sequence described in Table 1 can be determined conventionally using known computer programs such as the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wl 5371 1). Bestfit uses the local homology algorithm of Smith and Waterman (Advances in Applied Mathematics 2:482-489 ( 1981)), to find the best segment of homology between two sequences. When using Bestfit or any other sequence alignment program to determine whether a particular sequence is, for instance, 95% identical to a reference sequence according to the present invention, the parameters are set, of course, such that the percentage of identity is calculated over the full length of the reference nucleotide sequence and that gaps in homology of up to 5% of the total number of nucleotides in the reference sequence are allowed.
The present application is directed to nucleic acid molecules at least 90%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequences described in Table 1. One of skill in the art would still know how to use the nucleic acid molecule, for instance, as a hybridization probe or a polymerase chain reaction (PCR) primer. Uses of the nucleic acid molecules of the present invention include, inter alia, ( 1) isolating Streptococcal genes or allelic variants thereof from either a genomic or cDNA library and (2) Northern Blot or PCR analysis for detecting Streptococcal mRNA expression. Of course, due to the degeneracy of the genetic code, one of ordinary skill in the art will immediately recognize that a large number of nucleic acid molecules having a sequence at least 90%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence identified in Table 1 will encode the same polypeptide. In fact, since degenerate variants of these nucleotide sequences all encode the same polypeptide, this will be clear to the skilled artisan even without performing the above described comparison assay.
It will be further recognized in the art that, for such nucleic acid molecules that are not degenerate variants, a reasonable number will also encode proteins having antigenic epitopes of the S. pneumoniae polypeptides of the present invention. This is because the skilled artisan is fully aware of amino acid substitutions that are either less likely or not likely to significantly effect the antigenicity of a polypeptide (e.g. , replacement of an amino acid in a region which is not believed to form an antigenic epitope). For example, since antigenic epitopes have been identified which contain as few as six amino acids (see Harlow, et al, Antibodies: A Laboratory Manual, 2nd Ed.; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York (1988), page 76), in instances where a polypeptide has multiple antigenic epitopes the alteration of several amino acid residues would often not be expected to eliminate all of the antigenic epitopes of that polypeptide. This is especially so when the alterations are in regions believed to not constitute antigenic epitopes.
Vectors and Host Cells The present invention also relates to vectors which include the isolated
DNA molecules of the present invention, host cells which are genetically engineered with the recombinant vectors, and the production of S. pneumoniae polypeptides or fragments thereof by recombinant techniques.
Recombinant constructs may be introduced into host cells using well known techniques such as infection, transduction, transfection, transvection, electroporation and transformation. The vector may be, for example, a phage, plasmid, viral or retroviral vector. Retroviral vectors may be replication competent or replication defective. In the latter case, viral propagation generally will occur only in complementing host cells. The polynucleotides may be joined to a vector containing a selectable marker for propagation in a host. Generally, a plasmid vector is introduced in a precipitate, such as a calcium phosphate precipitate, or in a complex with a charged lipid. If the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells. Preferred are vectors comprising 's-acting control regions to the polynucleotide of interest. Appropriate frans-acting factors may be supplied by the host, supplied by a complementing vector or supplied by the vector itself upon introduction into the host.
In certain preferred embodiments in this regard, the vectors provide for specific expression, which may be inducible and/or cell type-specific.
Particularly preferred among such vectors are those inducible by environmental factors that are easy to manipulate, such as temperature and nutrient additives. Expression vectors useful in the present invention include chromosomal-, episomal- and virus-derived vectors, e.g. , vectors derived from bacterial plasmids, bacteriophage, yeast episomes, yeast chromosomal elements, viruses such as baculoviruses, papova viruses, vaccinia viruses, adenoviruses, fowl pox viruses, pseudorabies viruses and retroviruses, and vectors derived from combinations thereof, such as cosmids and phagemids.
The DNA insert should be operatively linked to an appropriate promoter, such as the phage lambda PL promoter, the E. coli lac, trp and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters will be known to the skilled artisan. The expression constructs will further contain sites for transcription initiation, termination and, in the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcripts expressed by the constructs will preferably include a translation initiating site at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated.
As indicated, the expression vectors will preferably include at least one selectable marker. Such markers include dihydrofolate reductase or neomycin resistance for eukaryotic cell culture and tetracycline or ampicillin resistance genes for culturing in E. coli and other bacteria. Representative examples of appropriate hosts include, but are not limited to, bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS and Bowes melanoma cells; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.
Among vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from Qiagen; pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNHlόa, pNH18A, pNH46A available from Stratagene; pET series of vectors available from Novagen; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRET5 available from Pharmacia. Among preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXTl and pSG available from Stratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Other suitable vectors will be readily apparent to the skilled artisan. Among known bacterial promoters suitable for use in the present invention include the E. coli lacl and lacZ promoters, the T3 and T7 promoters, the gpt promoter, the lambda PR and PL promoters and the trp promoter. Suitable eukaryotic promoters include the CMV immediate early promoter, the HSV thymidine kinase promoter, the early and late SV40 promoters, the promoters of retroviral LTRs, such as those of the Rous sarcoma virus (RSV), and metallothionein promoters, such as the mouse metallothionein-I promoter. Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection or other methods. Such methods are described in many standard laboratory manuals (for example. Davis, et al, Basic Methods In Molecular Biology (1986)).
Transcription of DNA encoding the polypeptides of the present invention by higher eukaryotes may be increased by inserting an enhancer sequence into the vector. Enhancers are cw-acting elements of DNA, usually about from 10 to 300 bp that act to increase transcriptional activity of a promoter in a given host cell-type. Examples of enhancers include the SV40 enhancer, which is located on the late side of the replication origin at bp 100 to 270, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.
For secretion of the translated polypeptide into the lumen of the endoplasmic reticulum, into the periplasmic space or into the extracellular environment, appropriate secretion signals may be incoφorated into the expressed polypeptide. The signals may be endogenous to the polypeptide or they may be heterologous signals.
The polypeptide may be expressed in a modified form, such as a fusion protein, and may include not only secretion signals, but also additional heterologous functional regions. For instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence in the host cell, during purification, or during subsequent handling and storage. Also, peptide moieties may be added to the polypeptide to facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to polypeptides to engender secretion or excretion, to improve stability and to facilitate purification, among others, are familiar and routine techniques in the art. A preferred fusion protein comprises a heterologous region from immunoglobulin that is useful to solubilize proteins. For example, EP-A-O 464 533 (Canadian counteφart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobin molecules together with another human protein or part thereof. In many cases, the Fc part in a fusion protein is thoroughly advantageous for use in therapy and diagnosis and thus results, for example, in improved pharmacokinetic properties (EP-A 0232 262). On the other hand, for some uses it would be desirable to be able to delete the Fc part after the fusion protein has been expressed, detected and purified in the advantageous manner described. This is the case when Fc portion proves to be a hindrance to use in therapy and diagnosis, for example when the fusion protein is to be used as antigen for immunizations. In drug discovery, for example, human proteins, such as, hEL5-receptor has been fused with Fc portions for the puφose of high-throughput screening assays to identify antagonists of hIL-5. See Bennett, D. et al, J. Molec. Recogn. 5:52-58 ( 1995) and Johanson, K. et al, J. Biol Chem. 270 (76,1:9459-9471 (1995). The S. pneumoniae polypeptides can be recovered and purified from recombinant cell cultures by well-known methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography, lectin chromatography and high performance liquid chromatography ("HPLC") is employed for purification. Polypeptides of the present invention include naturally purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect and mammalian cells.
Polypeptides and Fragments
The invention further provides isolated polypeptides having the amino acid sequences described in Table 1. and shown as SEQ ED NO:2, SEQ ED NO:4, SEQ ED NO:6, and so on through SEQ ID NO:226, and peptides or polypeptides comprising portions of the above polypeptides. The terms "peptide" and "oligopeptide" are considered synonymous (as is commonly recognized) and each term can be used interchangeably as the context requires to indicate a chain of at least two amino acids coupled by peptidyl linkages. The word "polypeptide" is used herein for chains containing more than ten amino acid residues. All oligopeptide and polypeptide formulas or sequences herein are written from left to right and in the direction from amino terminus to carboxy terminus.
Some amino acid sequences of the S. pneumoniae polypeptides described in Table 1 can be varied without significantly effecting the antigenicity of the polypeptides. If such differences in sequence are contemplated, it should be remembered that there will be critical areas on the polypeptide which determine antigenicity. In general, it is possible to replace residues which do not form part of an antigenic epitope without significantly effecting the antigenicity of a polypeptide. Guidance for such alterations is given in Table 2 wherein epitopes for each polypeptide is delineated.
The polypeptides of the present invention are preferably provided in an isolated form. By "isolated polypeptide" is intended a polypeptide removed from its native environment. Thus, a polypeptide produced and/or contained within a recombinant host cell is considered isolated for puφoses of the present invention. Also intended as an "isolated polypeptide" is a polypeptide that has been purified, partially or substantially, from a recombinant host cell. For example, recombinantly produced versions of the S. pneumoniae polypeptides described in Table 1 can be substantially purified by the one-step method described by Smith and Johnson (Gene 67:31-40 ( 1988)).
The polypeptides of the present invention include: (a) an amino acid sequence of any of the polypeptides described in Table 1 ; and (b) an amino acid sequence of an epitope-bearing portion of any one of the polypeptides of (a); as well as polypeptides with at least 70% similarity, and more preferably at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% similarity to those described in (a) or (b) above, as well as polypeptides having an amino acid sequence at least 70% identical, more preferably at least 75% identical, and still more preferably 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to those above.
By "% similarity" for two polypeptides is intended a similarity score produced by comparing the amino acid sequences of the two polypeptides using the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science
Drive, Madison, Wl 5371 1) and the default settings for determining similarity. Bestfit uses the local homology algorithm of Smith and Waterman (Advances in Applied Mathematics 2:482-489 ( 1981)) to find the best segment of similarity between two sequences. By a polypeptide having an amino acid sequence at least, for example,
95% "identical" to a reference amino acid sequence of a S. pneumoniae polypeptide is intended that the amino acid sequence of the polypeptide is identical to the reference sequence except that the polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the reference amino acid sequence. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a reference amino acid sequence, up to 5% of the amino acid residues in the reference sequence may be deleted or substituted with another amino acid, or a number of amino acids up to 5% of the total amino acid residues in the reference sequence may be inserted into the reference sequence. These alterations of the reference sequence may occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence.
The amino acid sequences shown in Table 1 may have on or more "X" residues. "X" represents unknown. Thus, for puφoses of defining identity, if any amino acid is present at the same position in a reference amino acid sequence (shown in Table 1) where an X is shown, the two sequences are identical at that position.
As a practical matter, whether any particular polypeptide is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to, for instance, an amino acid sequence shown in Table 1 , can be determined conventionally using known computer programs such the Bestfit program
(Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wl 5371 1). When using Bestfit or any other sequence alignment program to determine whether a particular sequence is, for instance, 95% identical to a reference sequence according to the present invention, the parameters are set, of course, such that the percentage of identity is calculated over the full length of the reference amino acid sequence and that gaps in homology of up to 5% of the total number of amino acid residues in the reference sequence are allowed.
As described below, the polypeptides of the present invention can also be used to raise polyclonal and monoclonal antibodies, which are useful in assays for detecting Streptococcal protein expression.
In another aspect, the invention provides peptides and polypeptides comprising epitope-bearing portions of the S. pneumoniae polypeptides of the invention. These epitopes are immunogenic or antigenic epitopes of the polypeptides of the invention. An "immunogenic epitope" is defined as a part of a protein that elicits an antibody response when the whole protein or polypeptide is the immunogen. These immunogenic epitopes are believed to be confined to a few loci on the molecule. On the other hand, a region of a protein molecule to which an antibody can bind is defined as an "antigenic determinant" or "antigenic epitope." The number of immunogenic epitopes of a protein generally is less than the number of antigenic epitopes (Gey sen, et al, Proc. Natl Acad. Sci. USA 81:3998- 4002 ( 1983)). Predicted antigenic epitopes are shown in Table 2, below. As to the selection of peptides or polypeptides bearing an antigenic epitope (i.e. , that contain a region of a protein molecule to which an antibody can bind), it is well known in that art that relatively short synthetic peptides that mimic part of a protein sequence are routinely capable of eliciting an antiserum that reacts with the partially mimicked protein (for instance, Sutcliffe, J., et al. ,
Science 219:660-666 ( 1983)). Peptides capable of eliciting protein-reactive sera are frequently represented in the primary sequence of a protein, can be characterized by a set of simple chemical rules, and are confined neither to immunodominant regions of intact proteins (i.e. , immunogenic epitopes) nor to the amino or carboxyl terminals. Peptides that are extremely hydrophobic and those of six or fewer residues generally are ineffective at inducing antibodies that bind to the mimicked protein; longer, peptides, especially those containing proline residues, usually are effective (Sutcliffe, et al , supra, p. 661). For instance, 18 of 20 peptides designed according to these guidelines, containing 8-39 residues covering 75% of the sequence of the influenza virus hemagglutinin HAl polypeptide chain, induced antibodies that reacted with the HA1 protein or intact virus; and 12/12 peptides from the MuLV polymerase and 18/18 from the rabies glycoprotein induced antibodies that precipitated the respective proteins. Antigenic epitope-bearing peptides and polypeptides of the invention are therefore useful to raise antibodies, including monoclonal antibodies, that bind specifically to a polypeptide of the invention. Thus, a high proportion of hybridomas obtained by fusion of spleen cells from donors immunized with an antigen epitope-bearing peptide generally secrete antibody reactive with the native protein (Sutcliffe, et al , supra, p. 663). The antibodies raised by antigenic epitope-bearing peptides or polypeptides are useful to detect the mimicked protein, and antibodies to different peptides may be used for tracking the fate of various regions of a protein precursor which undergoes post-translational processing. The peptides and anti-peptide antibodies may be used in a variety of qualitative or quantitative assays for the mimicked protein, for instance in competition assays since it has been shown that even short peptides (e.g., about 9 amino acids) can bind and displace the larger peptides in immunoprecipitation assays (for instance, Wilson, et al, Cell 37:767-778 ( 1984) p. 777). The anti-peptide antibodies of the invention also are useful for purification of the mimicked protein, for instance, by adsoφtion chromatography using methods well known in the art.
Antigenic epitope-bearing peptides and polypeptides of the invention designed according to the above guidelines preferably contain a sequence of at least seven, more preferably at least nine and most preferably between about 15 to about 30 amino acids contained within the amino acid sequence of a polypeptide of the invention. However, peptides or polypeptides comprising a larger portion of an amino acid sequence of a polypeptide of the invention, containing about 30 to about 50 amino acids, or any length up to and including the entire amino acid sequence of a polypeptide of the invention, also are considered epitope-bearing peptides or polypeptides of the invention and also are useful for inducing antibodies that react with the mimicked protein. Preferably, the amino acid sequence of the epitope-bearing peptide is selected to provide substantial solubility in aqueous solvents (i.e. , the sequence includes relatively hydrophilic residues and highly hydrophobic sequences are preferably avoided); and sequences containing proline residues are particularly preferred.
Non-limiting examples of antigenic polypeptides or peptides that can be used to generate Streptococcal-specific antibodies include portions of the amino acid sequences identified in Table 1. More specifically, Table 2 discloses antigenic fragments of polypeptides of the present invention, which antigenic fragments comprise amino acid sequences from about the first amino acid residues indicated to about the last amino acid residue indicated for each fragment. The polypeptide fragments disclosed in Table 2 are believed to be antigenic regions of the S. pneumoniae polypeptides described in Table 1. Thus the invention further includes isolated peptides and polypeptides comprising an amino acid sequence of an epitope shown in Table 2 and polynucleotides encoding said polypeptides.
The epitope-bearing peptides and polypeptides of the invention may be produced by any conventional means for making peptides or polypeptides including recombinant means using nucleic acid molecules of the invention. For instance, an epitope-bearing amino acid sequence of the present invention may be fused to a larger polypeptide which acts as a carrier during recombinant production and purification, as well as during immunization to produce anti-peptide antibodies. Epitope-bearing peptides also may be synthesized using known methods of chemical synthesis. For instance, Houghten has described a simple method for synthesis of large numbers of peptides, such as 10-20 mg of 248 different 13 residue peptides representing single amino acid variants of a segment of the HA1 polypeptide which were prepared and characterized (by ELISA-type binding studies) in less than four weeks (Houghten, R. A. Proc.
Natl. Acad. Sci. USA 82:5131-5135 ( 1985)). This "Simultaneous Multiple Peptide Synthesis (SMPS)" process is further described in U.S. Patent No. 4,631 ,21 1 to Houghten and coworkers ( 1986). In this procedure the individual resins for the solid-phase synthesis of various peptides are contained in separate solvent-permeable packets, enabling the optimal use of the many identical repetitive steps involved in solid-phase methods. A completely manual procedure allows 500-1000 or more syntheses to be conducted simultaneously (Houghten, et al, supra, p. 5134).
Epitope-bearing peptides and polypeptides of the invention are used to induce antibodies according to methods well known in the art (for instance, Sutcliffe, et al., supra; Wilson, et al, supra; Chow, M., et al, Proc. Natl. Acad. Sci. USA 82:910-914; and Bittle, F. J., et al, J. Gen. Virol 66:2347-2354 ( 1985)). Generally, animals may be immunized with free peptide; however, anti-peptide antibody titer may be boosted by coupling of the peptide to a macromolecular carrier, such as keyhole limpet hemacyanin (KLH) or tetanus toxoid. For instance, peptides containing cysteine may be coupled to carrier using a linker such as m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), while other peptides may be coupled to carrier using a more general linking agent such as glutaraldehyde. Animals such as rabbits, rats and mice are immunized with either free or carrier-coupled peptides, for instance, by intraperitoneal and/or intradermal injection of emulsions containing about 100 μg peptide or carrier protein and Freund's adjuvant. Several booster injections may be needed, for instance, at intervals of about two weeks, to provide a useful titer of anti-peptide antibody which can be detected, for example, by ELISA assay using free peptide adsorbed to a solid surface. The titer of anti-peptide antibodies in serum from an immunized animal may be increased by selection of anti-peptide antibodies, for instance, by adsoφtion to the peptide on a solid support and elution of the selected antibodies according to methods well known in the art.
Immunogenic epitope-bearing peptides of the invention, i.e., those parts of a protein that elicit an antibody response when the whole protein is the immunogen, are identified according to methods known in the art. For instance, Geysen, et al , supra, discloses a procedure for rapid concurrent synthesis on solid supports of hundreds of peptides of sufficient purity to react in an enzyme-linked immunosorbent assay. Interaction of synthesized peptides with antibodies is then easily detected without removing them from the support. In this manner a peptide bearing an immunogenic epitope of a desired protein may be identified routinely by one of ordinary skill in the art. For instance, the immunologically important epitope in the coat protein of foot-and-mouth disease virus was located by Geysen et al. supra with a resolution of seven amino acids by synthesis of an overlapping set of all 208 possible hexapeptides covering the entire 213 amino acid sequence of the protein. Then, a complete replacement set of peptides in which all 20 amino acids were substituted in turn at every position within the epitope were synthesized, and the particular amino acids conferring specificity for the reaction with antibody were determined. Thus, peptide analogs of the epitope-bearing peptides of the invention can be made routinely by this method. U.S. Patent No. 4,708,781 to Geysen (1987) further describes this method of identifying a peptide bearing an immunogenic epitope of a desired protein.
Further still, U.S. Patent No. 5, 194,392, to Geysen ( 1990), describes a general method of detecting or determining the sequence of monomers (amino acids or other compounds) which is a topological equivalent of the epitope (i.e. , a "mimotope") which is complementary to a particular paratope (antigen binding site) of an antibody of interest. More generally, U.S. Patent No. 4,433,092, also to Geysen ( 1989), describes a method of detecting or determining a sequence of monomers which is a topographical equivalent of a ligand which is complementary to the ligand binding site of a particular receptor of interest. Similarly, U.S. Patent No. 5,480,971 to Houghten, R. A. et al. (1996) discloses linear C -C -alkyl peralkylated oligopeptides and sets and libraries of such peptides, as well as methods for using such oligopeptide sets and libraries for determining the sequence of a peralkylated oligopeptide that preferentially binds to an acceptor molecule of interest. Thus, non-peptide analogs of the epitope-bearing peptides of the invention also can be made routinely by these methods.
The entire disclosure of each document cited in this section on "Polypeptides and Fragments" is hereby incoφorated herein by reference.
As one of skill in the art will appreciate, the polypeptides of the present invention and the epitope-bearing fragments thereof described above can be combined with parts of the constant domain of immunoglobulins (IgG), resulting in chimeric polypeptides. These fusion proteins facilitate purification and show an increased half-life in vivo. This has been shown, e.g. , for chimeric proteins consisting of the first two domains of the human CD4-polypeptide and various domains of the constant regions of the heavy or light chains of mammalian immunoglobulins (EPA 0,394,827; Traunecker et al, Nature 337:84-86 ( 1988)). Fusion proteins that have a disulfide-linked dimeric structure due to the IgG part can also be more efficient in binding and neutralizing other molecules than a monomeric S. pneumoniae polypeptide or fragment thereof alone (Fountoulakis et al, J. Biochem. 270:3958-3964 ( 1995)).
Diagnostic Assays The present invention further relates to a method for assaying for
Streptococcal infection in an animal via detecting the expression of genes encoding Streptococcal polypeptides (e.g., the polypeptides described Table 1 ). This method comprises analyzing tissue or body fluid from the animal for Streptococcus-specific antibodies or Streptococcal nucleic acids or proteins. Analysis of nucleic acid specific to Streptococcus can be done by PCR or hybridization techniques using nucleic acid sequences of the present invention as either hybridization probes or primers (cf. Molecular Cloning: A Laboratory Manual, second edition, edited by Sambrook, Fritsch, & Maniatis, Cold Spring Harbor Laboratory, 1989; Eremeeva et al, J. Clin. Microbiol 32:803-810 (1994) which describes differentiation among spotted fever group Rickettsiae species by analysis of restriction fragment length polymoφhism of PCR-amplified DNA). Methods for detecting B. burgdorferi nucleic acids via PCR are described, for example, in Chen et al, J. Clin. Microbiol. 32:589-595 ( 1994). Where diagnosis of a disease state related to infection with
Streptococcus has already been made, the present invention is useful for monitoring progression or regression of the disease state whereby patients exhibiting enhanced Streptococcus gene expression will experience a worse clinical outcome relative to patients expressing these gene(s) at a lower level. By "assaying for Streptococcal infection in an animal via detection of genes encoding Streptococcal polypeptides" is intended qualitatively or quantitatively measuring or estimating the level of one or more Streptococcus polypeptides or the level of nucleic acid encoding Streptococcus polypeptides in a first biological sample either directly (e.g. , by determining or estimating absolute protein level or nucleic level) or relatively (e.g. , by comparing to the
Streptococcus polypeptide level or mRNA level in a second biological sample). The Streptococcus polypeptide level or nucleic acid level in the second sample used for a relative comparison may be undetectable if obtained from an animal which is not infected with Streptococcus. When monitoring the progression or regression of a disease state, the Streptococcus polypeptide level or nucleic acid level may be compared to a second sample obtained from either an animal infected with Streptococcus or the same animal from which the first sample was obtained but taken from that animal at a different time than the first. As will be appreciated in the art, once a standard Streptococcus polypeptide level or nucleic acid level which corresponds to a particular stage of a Streptococcus infection is known, it can be used repeatedly as a standard for comparison.
By "biological sample" is intended any biological sample obtained from an animal, cell line, tissue culture, or other source which contains Streptococcus polypeptide, mRNA, or DNA. Biological samples include body fluids (such as plasma and synovial fluid) which contain Streptococcus polypeptides, and muscle, skin, and cartilage tissues. Methods for obtaining tissue biopsies and body fluids are well known in the art. The present invention is useful for detecting diseases related to
Streptococcus infections in animals. Preferred animals include monkeys, apes, cats, dogs, cows, pigs, mice, horses, rabbits and humans. Particularly preferred are humans.
Total RNA can be isolated from a biological sample using any suitable technique such as the single-step guanidinium-thiocyanate-phenol-chloroform method described in Chomczynski and Sacchi, Anal Biochem. 762: 156-159 ( 1987). mRNA encoding Streptococcus polypeptides having sufficient homology to the nucleic acid sequences identified in Table 1 to allow for hybridization between complementary sequences are then assayed using any appropriate method. These include Northern blot analysis, S I nuclease mapping, the polymerase chain reaction (PCR), reverse transcription in combination with the polymerase chain reaction (RT-PCR), and reverse transcription in combination with the ligase chain reaction (RT-LCR).
Northern blot analysis can be performed as described in Harada et al, Cell 63:303-312 ( 1990). Briefly, total RNA is prepared from a biological sample as described above. For the Northern blot, the RNA is denatured in an appropriate buffer (such as glyoxal/dimethyl sulfoxide/sodium phosphate buffer), subjected to agarose gel electrophoresis, and transferred onto a nitrocellulose filter. After the RNAs have been linked to the filter by a UV linker, the filter is prehybridized in a solution containing formamide, SSC,
Denhardt's solution, denatured salmon sperm, SDS, and sodium phosphate buffer. A S. pnuemoniae polypeptide DNA sequence shown in Table 1 labeled according to any appropriate method (such as the P-multiprimed DNA labeling system (Amersham)) is used as probe. After hybridization overnight, the filter is washed and exposed to x-ray film. DNA for use as probe according to the present invention is described in the sections above and will preferably at least 15 bp in length. S I mapping can be performed as described in Fujita et al, Cell 49:351-367 (1987). To prepare probe DNA for use in S I mapping, the sense strand of an above-described S. pnuemoniae DNA sequence of the present invention is used as a template to synthesize labeled antisense DNA. The antisense DNA can then be digested using an appropriate restriction endonuclease to generate further DNA probes of a desired length. Such antisense probes are useful for visualizing protected bands corresponding to the target mRNA (i.e., mRNA encoding Streptococcus polypeptides).
Preferably, levels of mRNA encoding Streptococcus polypeptides are assayed using the RT-PCR method described in Makino et al,
Technique 2:295-301 ( 1990). By this method, the radioactivities of the "amplicons" in the polyacrylamide gel bands are linearly related to the initial concentration of the target mRNA. Briefly, this method involves adding total RNA isolated from a biological sample in a reaction mixture containing a RT primer and appropriate buffer. After incubating for primer annealing, the mixture can be supplemented with a RT buffer, dNTPs, DTP, RNase inhibitor and reverse transcriptase. After incubation to achieve reverse transcription of the RNA, the RT products are then subject to PCR using labeled primers. Alternatively, rather than labeling the primers, a labeled dNTP can be included in the PCR reaction mixture. PCR amplification can be performed in a DNA thermal cycler according to conventional techniques. After a suitable number of rounds to achieve amplification, the PCR reaction mixture is electrophoresed on a polyacrylamide gel. After drying the gel, the radioactivity of the appropriate bands (corresponding to the mRNA encoding the Streptococcus polypeptides)) is quantified using an imaging analyzer. RT and PCR reaction ingredients and conditions, reagent and gel concentrations, and labeling methods are well known in the art. Variations on the RT-PCR method will be apparent to the skilled artisan.
Assaying Streptococcus polypeptide levels in a biological sample can occur using any art-known method. Preferred for assaying Streptococcus polypeptide levels in a biological sample are antibody-based techniques. For example, Streptococcus polypeptide expression in tissues can be studied with classical immunohistological methods. In these, the specific recognition is provided by the primary antibody (polyclonal or monoclonal) but the secondary detection system can utilize fluorescent, enzyme, or other conjugated secondary antibodies. As a result, an immunohistological staining of tissue section for pathological examination is obtained. Tissues can also be extracted, e.g. , with urea and neutral detergent, for the liberation of Streptococcus polypeptides for Western-blot or dot/slot assay (Jalkanen, M., et al, J. Cell Biol 707:976-985 (1985); Jalkanen, M., et al, J. Cell . Biol. 705:3087-3096 ( 1987)). In this technique, which is based on the use of cationic solid phases, quantitation of a Streptococcus polypeptide can be accomplished using an isolated Streptococcus polypeptide as a standard. This technique can also be applied to body fluids.
Other antibody-based methods useful for detecting Streptococcus polypeptide gene expression include immunoassays, such as the enzyme linked immunosorbent assay (ELISA) and the radioimmunoassay (RIA). For example, a Streptococcus polypeptide-specific monoclonal antibodies can be used both as an immunoabsorbent and as an enzyme-labeled probe to detect and quantify a Streptococcus polypeptide. The amount of a Streptococcus polypeptide present in the sample can be calculated by reference to the amount present in a standard preparation using a linear regression computer algorithm. Such an ELISA for detecting a tumor antigen is described in Iacobelli et al, Breast Cancer Research and Treatment 77: 19-30 ( 1988). In another ELISA assay, two distinct specific monoclonal antibodies can be used to detect Streptococcus polypeptides in a body fluid. In this assay, one of the antibodies is used as the immunoabsorbent and the other as the enzyme-labeled probe.
The above techniques may be conducted essentially as a "one-step" or "two-step" assay. The "one-step" assay involves contacting the Streptococcus polypeptide with immobilized antibody and, without washing, contacting the mixture with the labeled antibody. The "two-step" assay involves washing before contacting the mixture with the labeled antibody. Other conventional methods may also be employed as suitable. It is usually desirable to immobilize one component of the assay system on a support, thereby allowing other components of the system to be brought into contact with the component and readily removed from the sample.
Streptococcus polypeptide-specific antibodies for use in the present invention can be raised against an intact S. pneumoize polypeptide of the present invention or fragment thereof. These polypeptides and fragments may be administered to an animal (e.g. , rabbit or mouse) either with a carrier protein
(e.g. , albumin) or, if long enough (e.g., at least about 25 amino acids), without a carrier.
As used herein, the term "antibody" (Ab) or "monoclonal antibody" (Mab) is meant to include intact molecules as well as antibody fragments (such as, for example, Fab and F(ab') fragments) which are capable of specifically binding to a St rep tococcus polypeptide. Fab and F(ab')^ fragments lack the Fc fragment of intact antibody, clear more rapidly from the circulation, and may have less non-specific tissue binding of an intact antibody (Wahl et al, J. Nucl.
Med. 24:3 X6-325 (1983)). Thus, these fragments are preferred.
The antibodies of the present invention may be prepared by any of a variety of methods. For example, the S. pneumoniae polypeptides identified in Table 1 , or fragments thereof, can be administered to an animal in order to induce the production of sera containing polyclonal antibodies. In a preferred method, a preparation of a S. pneumoniae polypeptide of the present invention is prepared and purified to render it substantially free of natural contaminants.
Such a preparation is then introduced into an animal in order to produce polyclonal antisera of high specific activity.
In the most preferred method, the antibodies of the present invention are monoclonal antibodies. Such monoclonal antibodies can be prepared using hybridoma technology (Kohler et al, Nature 256:495 ( 1975); Kohler et al, Eur. J. Immunol. 6:51 1 ( 1976); Kohler et al, Eur. J. Immunol 6:292 ( 1976); Hammerling et al, In: Monoclonal Antibodies and T-Cell Hybridomas,
Elsevier, N.Y., ( 1981) pp. 563-681 ). In general, such procedures involve immunizing an animal (preferably a mouse) with a S. pneumoniae polypeptide antigen of the present invention. Suitable cells can be recognized by their capacity to bind ύ-Streptococcus polypeptide antibody. Such cells may be cultured in any suitable tissue culture medium; however, it is preferable to culture cells in Earle's modified Eagle's medium supplemented with 10% fetal bovine serum (inactivated at about 56°C), and supplemented with about 10 g/1 of nonessential amino acids, about 1 ,000 U/ml of penicillin, and about 100 μg/ml of streptomycin. The splenocytes of such mice are extracted and fused with a suitable myeloma cell line. Any suitable myeloma cell line may be employed in accordance with the present invention; however, it is preferable to employ the parent myeloma cell line (SP O), available from the American Type
Culture Collection, Rockville, Maryland. After fusion, the resulting hybridoma cells are selectively maintained in HAT medium, and then cloned by limiting dilution as described by Wands et al. (Gastroenterology 80:225-232 ( 1981)).
The hybridoma cells obtained through such a selection are then assayed to identify clones which secrete antibodies capable of binding the Streptococcus polypeptide antigen administered to immunized animal.
Alternatively, additional antibodies capable of binding to Streptococcus polypeptide antigens may be produced in a two-step procedure through the use of anti-idiotypic antibodies. Such a method makes use of the fact that antibodies are themselves antigens, and that, therefore, it is possible to obtain an antibody which binds to a second antibody. In accordance with this method,
Streptococcus polypeptide-specific antibodies are used to immunize an animal, preferably a mouse. The splenocytes of such an animal are then used to produce hybridoma cells, and the hybridoma cells are screened to identify clones which produce an antibody whose ability to bind to the Streptococcus polypeptide-specific antibody can be blocked by a Streptococcus polypeptide antigen. Such antibodies comprise anti-idiotypic antibodies to the Streptococcus polypeptide-specific antibody and can be used to immunize an animal to induce formation of further Streptococcus polypeptide-specific antibodies. It will be appreciated that Fab and F(ab') and other fragments of the antibodies of the present invention may be used according to the methods disclosed herein. Such fragments are typically produced by proteolytic cleavage, using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab') fragments). Alternatively, Streptococcus polypeptide-binding fragments can be produced through the application of recombinant DNA technology or through synthetic chemistry.
Of special interest to the present invention are antibodies to Streptococcus polypeptide antigens which are produced in humans, or are "humanized" (i.e. , non-immunogenic in a human) by recombinant or other technology. Humanized antibodies may be produced, for example by replacing an immunogenic portion of an antibody with a corresponding, but non- immunogenic portion (i.e. , chimeric antibodies) (Robinson, R.R. et al , International Patent Publication PCT/US86/02269; Akira, K. et al , European Patent Application 184, 187; Taniguchi, M., European Patent Application 171 ,496; Morrison, S.L. et al , European Patent Application 173,494;
Neuberger, M.S. et al., PCT Application WO 86/01533; Cabilly, S. et al , European Patent Application 125,023; Better, M. et al, Science 240: 1041-1043 (1988); Liu, A.Y. et al, Proc. Natl Acad. Sci. USA 84:3439-3443 (1987); Liu, A.Y. et al, J. Immunol. 739:3521-3526 (1987); Sun, L.K. et al, Proc. Natl. Acad. Sci. USA 84:214-218 (1987); Nishimura,
Y. et al, Cane. Res. 47:999-1005 (1987); Wood, C.R. et al, Nature 374:446-449 ( 1985)); Shaw et al, J. Natl Cancer Inst. 80: 1553-1559 ( 1988). Genera] reviews of "humanized" chimeric antibodies are provided by Morrison, S.L. (Science, 229: 1202-1207 ( 1985)) and by Oi, V.T. et al, BioTechniques 4:214 ( 1986)). Suitable "humanized" antibodies can be alternatively produced by CDR or CEA substitution (Jones, P.T. et al, Nature 327:552-525 ( 1986); Verhoeyan et al, Science 239: 1534 ( 1988); Beidler, C.B. et al, J. Immunol. 747:4053-4060 (1988)).
Suitable enzyme labels include, for example, those from the oxidase group, which catalyze the production of hydrogen peroxide by reacting with substrate. Glucose oxidase is particularly preferred as it has good stability and its substrate (glucose) is readily available. Activity of an oxidase label may be assayed by measuring the concentration of hydrogen peroxide formed by the enzyme-labeled antibody/substrate reaction. Besides enzymes, other suitable
125 121 14 labels include radioisotopes, such as iodine ( " I, I), carbon ( C), sulphur
,35 _ . . . 3T T . , . , 1 12, . . , . ,99m~ , r, ( S), tπtium ( H), indium ( In), and technetium ( Tc), and fluorescent labels, such as fluorescein and rhodamine, and biotin.
Further suitable labels for the Streptococcus polypeptide-specific antibodies of the present invention are provided below. Examples of suitable enzyme labels include malate dehydrogenase, staphylococcal nuclease, delta-5-steroid isomerase, yeast-alcohol dehydrogenase, alpha-glycerol phosphate dehydrogenase, triose phosphate isomerase, peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, beta-galactosidase, ribonuclease, urease, catalase, glucose-6-phosphate dehydrogenase, glucoamylase, and acetylcholine esterase. Examples of suitable radioisotopic labels include H, In, " I, I,
32„ 35^ 14 51 57m 58^ 59 75„ 152^ 90,, 67 _ 217 _,. 211 212„,
P, S, C, Cr, To, Co, Fe, Se, Eu, Y, Cu, Ci, At, Pb,
Sc, Pd. etc. In is a preferred isotope where in vivo imaging is used since its avoids the problem of dehalogenation of the I or I-labeled monoclonal antibody by the liver. In addition, this radionucleotide has a more favorable gamma emission energy for imaging (Perkins et al, Eur. J. Nucl Med.
70:296-301 (1985); Carasquillo et al, J. Nucl. Med. 28:281-287 ( 1987)). For example, In coupled to monoclonal antibodies with l-(P-isothiocyanatobenzyl)-DPTA has shown little uptake in non-tumorous tissues, particularly the liver, and therefore enhances specificity of tumor localization (Esteban et al, J. Nucl. Med. 28:861-870 (1987)).
Examples of suitable non-radioactive isotopic labels include Gd,
55 » . 162 τ^ 52^, , 56„
Mn, Dy, Tr, and Fe.
152
Examples of suitable fluorescent labels include an Eu label, a fluorescein label, an isothiocyanate label, a rhodamine label, a phycoerythrin label, a phycocyanin label, an allophycocyanin label, an o-phthaldehyde label, and a fluorescamine label.
Examples of suitable toxin labels include diphtheria toxin, ricin, and cholera toxin. Examples of chemiluminescent labels include a luminal label, an isoluminal label, an aromatic acridinium ester label, an imidazole label, an acridinium salt label, an oxalate ester label, a luciferin label, a luciferase label, and an aequorin label. Examples of nuclear magnetic resonance contrasting agents include heavy metal nuclei such as Gd, Mn, and iron.
Typical techniques for binding the above-described labels to antibodies are provided by Kennedy et al , Clin. Chim. Acta 70: 1-31 ( 1976), and Schurs et al, Clin. Chim. Acta 87: 1-40 (1977). Coupling techniques mentioned in the latter are the glutaraldehyde method, the periodate method, the dimaleimide method, the m-maleimidobenzyl-N-hydroxy-succinimide ester method, all of which methods are incoφorated by reference herein.
In a related aspect, the invention includes a diagnostic kit for use in screening serum containing antibodies specific against S. pneumoniae infection. Such a kit may include an isolated S. pneumoniae antigen comprising an epitope which is specifically immunoreactive with at least one anti-5. pneumoniae antibody. Such a kit also includes means for detecting the binding of said antibody to the antigen. In specific embodiments, the kit may include a recombinantly produced or chemically synthesized peptide or polypeptide antigen. The peptide or polypeptide antigen may be attached to a solid support.
In a more specific embodiment, the detecting means of the above- described kit includes a solid support to which said peptide or polypeptide antigen is attached. Such a kit may also include a non-attached reporter-labelled anti-human antibody. In this embodiment, binding of the antibody to the S . pneumoniae antigen can be detected by binding of the reporter labelled antibody to the anti-5. pneumoniae antibody.
In a related aspect, the invention includes a method of detecting S. pneumoniae infection in a subject. This detection method includes reacting a body fluid, preferrably serum, from the subject with an isolated S. pneumoniae antigen, and examining the antigen for the presence of bound antibody. In a specific embodiment, the method includes a polypeptide antigen attached to a solid support, and serum is reacted with the support. Subsequently, the support is reacted with a reporter-labelled anti-human antibody. The support is then examined for the presence of reporter-labelled antibody. The solid surface reagent employed in the above assays and kits is prepared by known techniques for attaching protein material to solid support material, such as polymeric beads, dip sticks, 96-well plates or filter material. These attachment methods generally include non-specific adsoφtion of the protein to the support or covalent attachment of the protein , typically through a free amine group, to a chemically reactive group on the solid support, such as an activated carboxyl, hydroxyl, or aldehyde group. Alternatively, streptavidin coated plates can be used in conjunction with biotinylated antigen(s).
Therapeutics and Modes of Administration
The present invention also provides vaccines comprising one or more polypeptides of the present invention. Heterogeneity in the composition of a vaccine may be provided by combining S. pneumoniae polypeptides of the present invention. Multi -component vaccines of this type are desirable because they are likely to be more effective in eliciting protective immune responses against multiple species and strains of the Streptococcus genus than single polypeptide vaccines. Thus, as discussed in detail below, a multi-component vaccine of the present invention may contain one or more, preferably 2 to about 20, more preferably 2 to about 15, and most preferably 3 to about 8, of the 5. pneumoniae pojypeptides identified in Table 1 , or fragments thereof.
Multi-component vaccines are known in the art to elicit antibody production to numerous immunogenic components. Decker, M. and Edwards, K., I. Infect. Dis. 774:S270-275 ( 1996). In addition, a hepatitis B, diphtheria, tetanus, pertussis tetravalent vaccine has recently been demonstrated to elicit protective levels of antibodies in human infants against all four pathogenic agents. Aristegui, J. et al, Vaccine 75:7-9 ( 1997).
The present invention thus also includes multi-component vaccines. These vaccines comprise more than one polypeptide, immunogen or antigen. An example of such a multi-component vaccine would be a vaccine comprising more than one of the S. pneumoniae polypeptides described in Table 1. A second example is a vaccine comprising one or more, for example 2 to 10, of the S. pneumoniae polypeptides identified in Table 1 and one or more, for example 2 to 10, additional polypeptides of either streptococcal or non-streptococcal origin. Thus, a multi-component vaccine which confers protective immunity to both a Streptococcal infection and infection by another pathogenic agent is also within the scope of the invention.
As indicated above, the vaccines of the present invention are expected to elicit a protective immune response against infections caused by species and strains of Streptococcus other than strain ofS. pneumoniae deposited with that
ATCC.
Further within the scope of the invention are whole cell and whole viral vaccines. Such vaccines may be produced recombinantly and involve the expression of one or more of the S. pneumoniae polypeptides described in Table 1. For example, the S. pneumoniae polypeptides of the present invention may be either secreted or localized intracellular, on the cell surface, or in the periplasmic space. Further, when a recombinant virus is used, the S. pneumoniae polypeptides of the present invention may, for example, be localized in the viral envelope, on the surface of the capsid, or internally within the capsid. Whole cells vaccines which employ cells expressing heterologous proteins are known in the art. See, e.g. , Robinson, K. et al, Nature Biotech. 75:653-657 (1997); Sirard, J. et al, Infect. Immun. 65:2029-2033 (1997); Chabalgoity, J. et al, Infect. Immun. 65:2402-2412 (1997). These cells may be administered live or may be killed prior to administration. Chabalgoity, J. et al , supra, for example, report the successful use in mice of a live attenuated Salmonella vaccine strain which expresses a portion of a platyhelminth fatty acid-binding protein as a fusion protein on its cells surface. A multi-component vaccine can also be prepared using techniques known in the art by combining one or more S. pneumoniae polypeptides of the present invention, or fragments thereof, with additional non-streptococcal components (e.g. , diphtheria toxin or tetanus toxin, and/or other compounds known to elicit an immune response). Such vaccines are useful for eliciting protective immune responses to both members of the Streptococcus genus and non-streptococcal pathogenic agents.
The vaccines of the present invention also include DNA vaccines. DNA vaccines are currently being developed for a number of infectious diseases. Boyer, J et al, Nat. Med. 3:526-532 (1997); reviewed in Spier, R., Vaccine 14: 1285-1288 (1996). Such DNA vaccines contain a nucleotide sequence encoding one or more S. pneumoniae polypeptides of the present invention oriented in a manner that allows for expression of the subject polypeptide. The direct administration of plasmid DNA encoding B. burgdorgeri OspA has been shown to elicit protective immunity in mice against borrelial challenge. Luke, C. et al, J. Infect. Dis. 775:91-97 ( 1997).
The present invention also relates to the administration of a vaccine which is co-administered with a molecule capable of modulating immune responses. Kim, J. et al, Nature Biotech. 75:641-646 ( 1997), for example, report the enhancement of immune responses produced by DNA immunizations when DNA sequences encoding molecules which stimulate the immune response are co-administered. In a similar fashion, the vaccines of the present invention may be co-administered with either nucleic acids encoding immune modulators or the immune modulators themselves. These immune modulators include granulocyte macrophage colony stimulating factor (GM-CSF) and CD86.
The vaccines of the present invention may be used to confer resistance to streptococcal infection by either passive or active immunization. When the vaccines of the present invention are used to confer resistance to streptococcal infection through active immunization, a vaccine of the present invention is administered to an animal to elicit a protective immune response which either prevents or attenuates a streptococcal infection. When the vaccines of the present invention are used to confer resistance to streptococcal infection through passive immunization, the vaccine is provided to a host animal (e.g. , human, dog, or mouse), and the antisera elicited by this antisera is recovered and directly provided to a recipient suspected of having an infection caused by a member of the Streptococcus genus.
The ability to label antibodies, or fragments of antibodies, with toxin molecules provides an additional method for treating streptococcal infections when passive immunization is conducted. In this embodiment, antibodies, or fragments of antibodies, capable of recognizing the 5. pneumoniae polypeptides disclosed herein, or fragments thereof, as well as other Streptococcus proteins, are labeled with toxin molecules prior to their administration to the patient. When such toxin derivatized antibodies bind to Streptococcus cells, toxin moieties will be localized to these cells and will cause their death.
The present invention thus concerns and provides a means for preventing or attenuating a streptococcal infection resulting from organisms which have antigens that are recognized and bound by antisera produced in response to the polypeptides of the present invention. As used herein, a vaccine is said to prevent or attenuate a disease if its administration to an animal results either in the total or partial attenuation (i.e. , suppression) of a symptom or condition of the disease, or in the total or partial immunity of the animal to the disease. The administration of the vaccine (or the antisera which it elicits) may be for either a "prophylactic" or "therapeutic" puφose. When provided prophylactically, the compound(s) are provided in advance of any symptoms of streptococcal infection. The prophylactic administration of the compound(s) serves to prevent or attenuate any subsequent infection. When provided therapeutically, the compound(s) is provided upon or after the detection of symptoms which indicate that an animal may be infected with a member of the Streptococcus genus. The therapeutic administration of the compound(s) serves to attenuate any actual infection. Thus, the S. pneumoniae polypeptides, and fragments thereof, of the present invention may be provided either prior to the onset of infection (so as to prevent or attenuate an anticipated infection) or after the initiation of an actual infection.
The polypeptides of the invention, whether encoding a portion of a native protein or a functional derivative thereof, may be administered in pure form or may be coupled to a macromolecular carrier. Example of such carriers are proteins and carbohydrates. Suitable proteins which may act as macromolecular carrier for enhancing the immunogenicity of the polypeptides of the present invention include keyhole limpet hemacyanin (KLH) tetanus toxoid, pertussis toxin, bovine serum albumin, and ovalbumin. Methods for coupling the polypeptides of the present invention to such macromolecular carriers are disclosed in Harlow et al, Antibodies: A Laboratory Manual, 2nd Ed.; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York ( 1988), the entire disclosure of which is incoφorated by reference herein. A composition is said to be "pharmacologically acceptable" if its administration pan be tolerated by a recipient animal and is otherwise suitable for administration to that animal. Such an agent is said to be administered in a "therapeutically effective amount" if the amount administered is physiologically significant. An agent is physiologically significant if its presence results in a detectable change in the physiology of a recipient patient.
While in all instances the vaccine of the present invention is administered as a pharmacologically acceptable compound, one skilled in the art would recognize that the composition of a pharmacologically acceptable compound varies with the animal to which it is administered. For example, a vaccine intended for human use will generally not be co-administered with Freund's adjuvant. Further, the level of purity of the S. pneumoniae polypeptides of the present invention will normally be higher when administered to a human than when administered to a non-human animal.
As would be understood by one of ordinary skill in the art, when the vaccine of the present invention is provided to an animal, it may be in a composition which may contain salts, buffers, adjuvants, or other substances which are desirable for improving the efficacy of the composition. Adjuvants are substances that can be used to specifically augment a specific immune response. These substances generally perform two functions: ( 1) they protect the antigen(s) from being rapidly catabolized after administration and (2) they nonspecifically stimulate immune responses.
Normally, the adjuvant and the composition are mixed prior to presentation to the immune system, or presented separately, but into the same site of the animal being immunized. Adjuvants can be loosely divided into several groups based upon their composition. These groups include oil adjuvants (for example, Freund's complete and incomplete), mineral salts (for example, AlK(SO ) AlNa(SO \, A1NH (SO ), silica, kaolin, and carbon), polynucleotides (for example, poly IC and poly AU acids), and certain natural substances (for example, wax D from Mycobacterium tuberculosis, as well as substances found in Corynebacterium parvum, or Bordetella pertussis, and members of the genus Brucella. Other substances useful as adjuvants are the saponins such as, for example, Quil A. (Superfos A/S, Denmark). Preferred adjuvants for use in the present invention include aluminum salts, such as
AlK(SO 4 ) 2 , AlNa(SO 4 ) 2 , and A1NH 4 (SO 4 ). Examples of materials suitable for use in vaccine compositions are provided in Remington's Pharmaceutical Sciences (Osol, A, Ed, Mack Publishing Co, Easton, PA, pp. 1324-1341 (1980), which reference is incoφorated herein by reference). The therapeutic compositions of the present invention can be administered parenterally by injection, rapid infusion, nasopharyngeal absoφtion (intranasopharangeally), dermoabsoφtion, or orally. The compositions may alternatively be administered intramuscularly, or intravenously. Compositions for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Carriers or occlusive dressings can be used to increase skin permeability and enhance antigen absoφtion. Liquid dosage forms for oral administration may generally comprise a liposome solution containing the liquid dosage form. Suitable forms for suspending liposomes include emulsions, suspensions, solutions, syrups, and elixirs containing inert diluents commonly used in the art, such as purified water. Besides the inert diluents, such compositions can also include adjuvants, wetting agents, emulsifying and suspending agents, or sweetening, flavoring, or perfuming agents.
Therapeutic compositions of the present invention can also be administered in encapsulated form. For example, intranasal immunization of mice against Bordetella pertussis infection using vaccines encapsulated in biodegradable microsphere composed of poly(DL-lactide-co-glycolide) has been shown to stimulate protective immune responses. Shahin, R. et al., Infect.
Immun. 63: 1195-1200 ( 1995). Similarly, orally administered encapsulated Salmonella typhimurium antigens have also been shown to elicit protective immunity in mice. Allaoui-Attarki, K. et al, Infect. Immun. 65:853-857 (1997). Encapsulated vaccines of the present invention can be administered by a variety of routes including those involving contacting the vaccine with mucous membranes (e.g., intranasally, intracolonicly, intraduodenally). Many different techniques exist for the timing of the immunizations when a multiple administration regimen is utilized. It is possible to use the compositions of the invention more than once to increase the levels and diversities of expression of the immunoglobulin repertoire expressed by the immunized animal. Typically, if multiple immunizations are given, they will be given one to two months apart.
According to the present invention, an "effective amount" of a therapeutic composition is one which is sufficient to achieve a desired biological effect. Generally, the dosage needed to provide an effective amount of the composition will vary depending upon such factors as the animal's or human's age, condition, sex, and extent of disease, if any, and other variables which can be adjusted by pne of ordinary skill in the art.
The antigenic preparations of the invention can be administered by either single or multiple dosages of an effective amount. Effective amounts of the compositions of the invention can vary from 0.01- 1 ,000 μg/ml per dose, more preferably 0.1-500 μg/ml per dose, and most preferably 10-300 μg/ml per dose.
Having now generally described the invention, the same will be more readily understood through reference to the following example which is provided by way of illustration, and is not intended to be limiting of the present invention, unless specified.
Examples
Example 1: Expression and Purification of S. pneumoniae
Polypeptides in E. coli The bacterial expression vector pQElO (QIAGEN, Inc., 9259 Eton
Avenue, Chatsworth, CA, 9131 1) is used in this example for cloning of the nucleotide sequences shown in Table 1 and for expressing the polypeptides identified in Table 1. The components of the pQElO plasmid are arranged such that the inserted DNA sequence encoding a polypeptide of the present invention expresses the polypeptide with the six His residues (i. e. , a "6 X His tag")) covalently linked to the amino terminus.
The DNA sequences encoding the desired portions of the polypeptides of Table 1 are amplified using PCR oligonucleotide primers from either a DNA library constructed from S. pnuemonicae, such as the one deposited by the inventors at the ATCC for convenience, ATCC Deposit No. 97755, or from DNA isolated from the same organism such as the S. pneumoniae strain deposited with the ATCC as Deposit No. 55840. A list of PCR primers which can be used for this puφose is provided in Table 3, below. The PCR primers anneal to the nucleotide sequences encoding both the amino terminal and carboxy terminal amino acid sequences of the desired portion of the polypeptides of Table 1. Additional nucleotides containing restriction sites to facilitate cloning in the pQElO vector were added to the 5' and 3' primer sequences, respectively. Such restriction sites are listed in Table 3 for each primer. In each case, the primer comprises, from the 5' end, 4 random nucleotides to prevent "breathing" during the annealing process, a restriction site (shown in Table 3), and approximately 15 nucleotides of S. pneumoniae ORF sequence (the complete sequence of each cloning primer is shown as SEQ ED NO:227 through SEQ ID NO:452).
For cloning the polypeptides of Table 1 , the 5' and 3' primers were selected to amplify their respective nucleotide coding sequences. One of ordinary skill in the art would appreciate that the point in the protein coding sequence where the 5' primer begins may be varied to amplify a DNA segment encoding any desired portion of the complete amino acid sequences described in
Table 1. Similarly, one of ordinary skill in the art would further appreciate that the point in the protein coding sequence where the 3' primer begins may also be varied to amplify a DNA segment encoding any desired portion of the complete amino acid sequences described in Table 1. The amplified DNA fragment and the pQElO vector are digested with the appropriate restriction enzyme(s) and the digested DNAs are then ligated together. The ligation mixture is transformed into competent E. coli cells using standard procedures such as those described in Sambrook et al, Molecular Cloning: a Laboratory Manual, 2nd Ed.; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. ( 1989). Transformants are identified by their ability to grow under selective pressure on LB plates. Plasmid DNA is isolated from resistant colonies and the identity of the cloned DNA confirmed by restriction analysis, PCR and DNA sequencing.
Clones containing the desired constructs are grown overnight ("O/N") in liquid culture under selection. The O/N culture is used to inoculate a large culture, at a dilution of approximately 1:25 to 1 :250. The cells are grown to an optical density at 600 nm ("OD600") of between 0.4 and 0.6. Isopropyl-b-D- thiogalactopyranoside ("IPTG") is then added to a final concentration of 1 mM to induce transcription from the lac repressor sensitive promoter, by inactivating the lacl repressor. Cells subsequently are incubated further for 3 to 4 hours. Cells are then harvested by centrifugation.
The cells are stirred for 3-4 hours at 4 C in 6M guanidine-HCl, pH 8. The cell debris is removed by centrifugation, and the supernatant containing the protein of interest is loaded onto a nickel-nitrilo-tri-acetic acid ("NiNTA") affinity resin column (available from QIAGEN, Inc., supra). Proteins with a 6x His tag bind to the NI-NTA resin with high affinity and can be purified in a simple one-step procedure (for details see: The QIAexpressionist, 1995, QIAGEN, Inc., supra). Briefly, the supernatant is loaded onto the column in 6
M guanidine-HCl, pH8, the column is first washed with 10 volumes of 6 M guanidine-HCl, pH8, then washed with 10 volumes of 6 M guanidine-HCl pH6, and finally the polypeptide is eluted with 6 M guanidine-HCl, pH 5.0.
The purified protein is then renatured by dialyzing it against phosphate - buffered saline (PBS) or 50 mM Na-acetate, pH 6 buffer plus 200 mM NaCl.
Alternatively, the protein can be successfully refolded while immobilized on the Ni-NTA column. The recommended conditions are as follows: renature using a linear 6M-1M urea gradient in 500 mM NaCl, 20% glycerol, 20 mM Tris/HCl pH7.4, containing protease inhibitors. The renaturation should be performed over a period of 1.5 hours or more. After renaturation the proteins can be eluted by the addition of 250 mM imidazole. Imidazole is removed by a final dialyzing step against PBS or 50 mM sodium acetate pH6 buffer plus 200 mM NaCl. The purified protein is stored at 4°C or frozen at -80°C.
The DNA sequences encoding the amino acid sequences of Table 1 may also be cloned and expressed as fusion proteins by a protocol similar to that described directly above, wherein the pET-32b(+) vector (Novagen, 601 Science Drive, Madison, Wl 53711) is preferentially used in place of pQElO.
Each of the polynucleotides shown in Table 1 , was successfully amplified and subcloned into pQElO as described above using the PCR primers shown in Table 3. These pQElO plasmids containing the DNAs of Table 1 , except SP023, SP042, SP054, SP063, SP081 , SP092, SPI 14, SP122, SP123, SP126, and SP127, were deposited with the ATCC as a pooled deposit as a convenience to those of skill in the art. This pooled deposit was desposited on October 16, 1997 and given ATCC Deposit No. 209369. Those of ordinary skill in the art appreciate that isolating an individual plasmid from the pooled deposit is trivial provided the information and reagents described herein. Each of the deposited clones is capable of expressing its encoded S. pneumoniae polypeptide. Example 2: Immunization and Detection of Immune Responses
Methods
Growth of bacterial innoculum, immunization of Mice and Challenge with S pneumoniae.
Propagation and storage of, and challenge by S. pneumoniae are preformed essentially as described in Aaberge, LS. et al., Virulence of Streptococcus pneumoniae in mice: a standardized method for preparation and frozen storage of the experimental bacterial inoculum, Microbial Pathogenesis, 18: 141 (1995), incoφorated herein by reference.
Briefly, Todd Hewitt (TH) broth (Difco laboratories, Detroit, MI) with 17% FCS, and horse blood agar plates are used for culturing the bacteria. Both broth and blood plates are incubated at 37°C in a 5% CO2 atmosphere. Blood plates are incubated for 18 hr. The culture broth is regularly 10-fold serially diluted in TH broth kept at room temperature and bacterial suspensions are kept at room temperature until challenge of mice.
For active immunizations C3H7HeJ mice (The Jackson Laboratory, Bar Harbor, ME) are injected intraperitoneally (i.p.) at week 0 with 20 g of recombinant streptococcal protein, or phosphate-buffered saline (PBS), emulsified with complete Freund's adjuvant (CFA), given a similar booster immunization in incomplete Freund's adjuvant (EFA) at week 4, and challenged at week 6. For challenge S. pneumoniae are diluted in TH broth from exponentially-growing cultures and mice are injected subcutaneously (s.c.) at the base of the tail with 0.1 ml of these dilutions (serial dilutions are used to find medium infectious dose). Streptococci used for challenge are passaged fewer than six times in vitro. To assess infection, blood samples are obtained from the distal part of the lateral femoral vein into heparinized capillary tubes. A 25 ul blood sample is serially 10-fold diluted in TH broth, and 25 ul of diluted and undiluted blood is plated onto blood agar plates. The plates are incubated for 18 hr. and colonies are counted.
Other methods are known in the art, for example, see Langermann, S. et al., J. Exp. Med., 180:2277 (1994), incoφorated herein by reference. Immunoassays
Several immunoassay formats are used to quantify levels of streptococcal-specific antibodies (ELISA and immunoblot), and to evaluate the functional properties of these antibodies (growth inhibition assay). The ELISA and immunoblot assays are also used to detect and quantify antibodies elicited in response to streptococcal infection that react with specific streptococcal antigens. Where antibodies to certain streptococcal antigens are elicited by infection this is taken as evidence that the streptococcal proteins in question are expressed in vivo. Absence of infection-derived antibodies (seroconversion) following streptococcal challenge is evidence that infection is prevented or suppressed. The immunoblot assay is also used to ascertain whether antibodies raised against recombinant streptococcal antigens recognize a protein of similar size in extracts of whole streptococci. Where the natural protein is of similar, or identical, size in the immunoblot assay to the recombinant version of the same protein, this is taken as evidence that the recombinant protein is the product of a full-length clone of the respective gene.
Enzyme-Linked Immunosorbant Assay (ELISA).
The ELISA is used to quantify levels of antibodies reactive with streptococcus antigens elicited in response to immunization with these streptococcal antigens.
Wells of 96 well microtiter plates (Immunlon 4, Dynatech, Chantilly, Virginia, or equivalent) are coated with antigen by incubating 50 1 of 1 g/ml protein antigen solution in a suitable buffer, typically 0.1 M sodium carbonate buffer at pH 9.6. After decanting unbound antigen, additional binding sites are blocked by incubating 100 1 of 3% nonfat milk in wash buffer (PBS, 0.2% Tween 20, pH 7.4). After washing, duplicate serial two-fold dilutions of sera in PBS , Tween 20, 1 % fetal bovine serum, are incubated for 1 hr, removed, wells are washed three times, and incubated with horseradish peroxidase-conjugated goat anti-mouse IgG. After three washes, bound antibodies are detected with H2O2 and 2,2'-azino-di-(3-ethylbenzthiazoline sulfonate) (Schwan, T.G., et al, Proc.
Natl. Acad. Sci. USA 92:2909-2913 ( 1985)) (ABTS®, Kirkegaard & Perry Labs., Gaithersburg, MD) and A405 is quantified with a Molecular Devices,
Coφ. (Menlo Park, California) Vmax™ plate reader. IgG levels twice the background level in serum from naive mice are assigned the minimum titer of 1 : 100. Sodiumdodecylsulfate-Poly acrylamide Gel Electrophoresis
(SDS-PAGE) and Immunoblotting
Using a single well format, total streptococcal protein extracts or recombinant streptococcal antigen are boiled in SDS/2-ME sample buffer before electrophoresis through 3% acrylamide stacking gels, and resolving gels of higher acrylamide concentration, typically 10-15% acrylamide monomer. Gels are electro-blotted to nitrocellulose membranes and lanes are probed with dilutions of antibody to be tested for reactivity with specific streptococcal antigens, followed by the appropriate secondary antibody-enzyme (horseradish peroxidase) conjugate. When it is desirable to confirm that the protein had transferred following electro-blotting, membranes are stained with Ponceau S . Immunoblot signals from bound antibodies are detected on x-ray film as chemiluminescence using ECL™ reagents (Amersham Coφ., Arlington Heights, Illinois).
/ Example 3: Detection of Streptococcus mRNA expression
Northern blot analysis is carried out using methods described by, among others, Sambrook et al, supra, to detect the expression of the S. pneumoniae nucleotide sequences of the present invention in animal tissues. A cDNA probe containing an entire nucleotide sequence shown in Table 1 is labeled with 32p using the redipήme™ DNA labeling system (Amersham Life Science), according to manufacturer's instructions. After labeling, the probe is purified using a CHROMA SPIN- 100™ column (Clontech Laboratories, Inc.), according to manufacturer's protocol number PT 1200- 1. The purified labeled probe is then used to detect the expression of Streptococcus mRNA in an animal tissue sample.
Animal tissues, such as blood or spinal fluid, are examined with the labeled probe using ExpressHyb™ hybridization solution (Clontech) according to manufacturer's protocol number PT 1 190- 1. Following hybridization and washing, the blots are mounted and exposed to film at -70 C overnight, and films developed according to standard procedures. It will be clear that the invention may be practiced otherwise than as particularly described in the foregoing description and examples.
Numerous modifications and variations of the present invention are possible in light of the above teachings and, therefore, are within the scope of the appended claims.
The entire disclosure of all publications (including patents, patent applications, journal articles, laboratory manuals, books, or other documents) cited herein are hereby incoφorated by reference.
Table 1 48
SP001 nucleotide (SEQ ID NO:l)
TAAAATCTACGACAATAAAAATCAACTCATTGCTGACTTGGGTTCTGAACGCCGCGTCAATGCCCAAGC TAATGATATTCCCACAGATTTGGTTAAGGCAATCGTTTCTATCGAAGACCATCGCTTCTTCGACCACAG GGGGATTGATACCATCCGTATCCTGGGAGCTTTCTTGCGCAATCTGCAAAGCAATTCCCTCCAAGGTGG ATCAACTCTCACCCAACAGTTGATTAAGTTGACTTACTTTTCAACTTCGACTTCCGACCAGACTATTTC TCGTAAGGCTCAGGAAGCTTGGTTAGCGATTCAGTTAGAACAAAAAGCAACCAAGCAAGAAATCTTGAC CTACTATATAAATAAGGTCTACATGTCTAATGGGAACTATGGAATGCAGACAGCAGCTCAAAACTACTA TGGTAAAGACCTCAATAATTTAAGTTTACCTCAGTTAGCCTTGCTGGCTGGAATGCCTCAGGCACCAAA CCAATATGACCCCTATTCACATCCAGAAGCAGCCCAAGACCGCCGAAACTTGGTCTTATCTGAAATGAA AAATCAAGGCTACATCTCTGCTGAACAGTATGAGAAAGCAGTCAATACACCAATTACTGATGGACTACA AAGTCTCAAATCAGCAAGTAATTACCCTGCTTACATGGATAATTACCTCAAGGAAGTCATCAATCAAGT TGAAGAAGAAACAGGCTATAACCTACTCACAACTGGGATGGATGTCTACACAAATGTAGACCAAGAAGC TCAAAAACATCTGTGGGATATTTACAATACAGACGAATACGTTGCCTATCCAGACGATGAATTGCAAGT CGCTTCTACCATTGTTGATGTTTCTAACGGTAAAGTCATTGCCCAGCTAGGAGCACGCCATCAGTCAAG TAATGTTTCCTTCGGAATTAACCAAGCAGTAGAAACAAACCGCGACTGGGGATCAACTATGAAACCGAT CACAGACTATGCTCCTGCCTTGGAGTACGGTGTCTACGATTCAACTGCTACTATCGTTCACGATGAGCC CTATAACTACCCTGGGACAAATACTCCTGTTTATAACTGGGATAGGGGCTACTTTGGCAACATCACCTT GCAATACGCCCTGCAACAATCGCGAAACGTCCCAGCCGTGGAAACTCTAAACAAGGTCGGACTCAACCG CGCCAAGACTTTCCTAAATGGTCTAGGAATCGACTACCCAAGTATTCACTACTCAAATGCCATTTCAAG TAACACAACCGAATCAGACAAAAAATATGGAGCAAGTAGTGAAAAGATGGCTGCTGCTTACGCTGCCTT TGCAAATGGTGGAACTTACTATAAACCAATGTATATCCATAAAGTCGTCTTTAGTGATGGGAGTGAAAA AGAGTTCTCTAATGTCGGAACTCGTGCCATGAAGGAAACGACAGCCTATATGATGACCGACATGATGAA AACAGTCTTGACTTATGGAACTGGACGAAATGCCTATCTTGCTTGGCTCCCTCAGGCTGGTAAAACAGG AACCTCTAACTATACAGACGAGGAAATTGAAAACCACATCAAGACCTCTCAATTTGTAGCACCTGATGA ACTATTTGCTGGCTATACGCGTAAATATTCAATGGCTGTATGGACAGGCTATTCTAACCGTCTGACACC ACTTGTAGGCAATGGCCTTACGGTCGCTGCCAAAGTTTACCGCTCTATGATGACCTACCTGTCTGAAGG AAGCAATCCAGAAGATTGGAATATACCAGAGGGGCTCTACAGAAATGGAGAATTCGTATTTAAAAATGG TGCTCGTTCTACGTGGAACTCACCTGCTCCACAACAACCCCCATCAACTGAAAGTTCAAGCTCATCATC AGATAGTTCAACTTCACAGTCTAGCTCAACCACTCCAAGCACAAATAATAGTACGACTACCAATCCTAA CAATAATACGCAACAATCAAATACAACCCCTGATCAACAAAATCAGAATCCTCAACCAGCACAACCA
SP001 AMINO ACID (SEQ ID NO : 2 )
KIYDNKNQLIADLGSERRVNAQANDIPTDLVKAIVΞIEDHRFFDHRGIDTIRILGAFLKNLQSNΞLQGG STLTQQLIK TYFSTSTSDQTISRKAQEA LAIQLEQKATKQEILTYYINKVΎMSNGNYGMQTAAQNYY GKDLNNLSLPQLALLAGMPQAPNQYDPYSHPEAAQDRRNLVLSEMK QGYISAEQYEKAVNTPITDGLQ SLKSASNYPAYMDNYLKEVINQVEEETGYNLLTTGMDVYTNVDQEAQKHL DIYNTDEYVAYPDDELQV AΞTIVDVSNGKVIAQLGARHQSΞNVSFGINQAVETNRD GSTMKPITDYAPALEYGVYDSTATIVHDEP YNYPGTNTPVYNWDRGYFGNIT QYALQQSRNVPAVETLNKVGLNRAKTFLNGLGIDYPSIHYSNAISS NTTESDKKYGASSEK AAAYAAFANGGTYYKPMYIHKVVFSDGSEKΞFSNVGTRAMKETTAYMMTD MK TVLTYGTGRNAYLA LPQAGKTGTSNYTDEEIENHIKTSQFVAPDELFAGYTRKYSMAV TGYSNRLTP LVGNGLTVAAKVYRS MTYLSEGSNPEDWNIPEGLYRNGEFVFKNGARSTWNSPAPQQPPSTESSSSSS DSSTSQSSΞTTPSTNNSTTTNP TQQSNTTPDQQNQNPQPAQP
SP004 nucleotide (SEQ ID NO : 3 )
AAATTACAATACGGACTATGAATTGACCTCTGGAGAAAAATTACCTCTTCCTAAAGAGATTTCAGGTTA CACTTATATTGGATATATCAAAGAGGGAAAAACGACTTCTGAGTCTGAAGTAAGTAATCAAAAGAGTTC AGTTGCCACTCCTACAAAACAACAAAAGGTGGATTATAATGTTACACCGAATTTTGTAGACCATCCATC AACAGTACAAGCTATTCAGGAACAAACACCTGTTTCTTCAACTAAGCCGACAGAAGTTCAAGTAGTTGA AAAACCTTTCTCTACTGAATTAATCAATCCAAGAAAAGAAGAGAAACAATCTTCAGATTCTCAAGAACA ATTAGCCGAACATAAGAATCTAGAAACGAAGAAAGAGGAGAAGATTTCTCCAAAAGAAAAGACTGGGGT AAATACATTAAATCCACAGGATGAAGTTTTATCAGGTCAATTGAACAAACCTGAACTCTTATATCGTGA GGAAACTATGGAGACAAAAATAGATTTTCAAGAAGAAATTCAAGAAAATCCTGATTTAGCTGAAGGAAC TGTAAGAGTAAAACAAGAAGGTAAATTAGGTAAGAAAGTTGAAATCGTCAGAATATTCTCTGTAAACAA GGAAGAAGTTTCGCGAGAAATTGTTTCAACTTCAACGACTGCGCCTAGTCCAAGAATAGTCGAAAAAGG TACTAAAAAAACTCAAGTTATAAAGGAACAACCTGAGACTGGTGTAGAACATAAGGACGTACAGTCTGG AGCTATTGTTGAACCCGCAATTCAGCCTGAGTTGCCCGAAGCTGTAGTAAGTGACAAAGGCGAACCAGA AGTTCAACCTACATTACCCGAAGCAGTTGTGACCGACAAAGGTGAGACTGAGGTTCAACCAGAGTCGCC AGATACTGTGGTAAGTGATAAAGGTGAACCAGAGCAGGTAGCACCGCTTCCAGAATATAAGGGTAATAT Table 1 49
TGAGCAAGTAAAACCTGAAACTCCGGTTGAGAAGACCAAAGAACAAGGTCCAGAAAAAACTGAAGAAGT TCCAGTAAAACCAACAGAAGAAACACCAGTAAATCCAAATGAAGGTACTACAGAAGGAACCTCAATTCA AGAAGCAGAAAATCCAGTTCAACCTGCAGAAGAATCAACAACGAATTCAGAGAAAGTATCACCAGATAC ATCTAGCAAAAATACTGGGGAAGTGTCCAGTAATCCTAGTGATTCGACAACCTCAGTTGGAGAATCAAA TAAACCAGAACATAATGACTCTAAAAATGAAAATTCAGAAAAAACTGTAGAAGAAGTTCCAGTAAATCC AAATGAAGGCACAGTAGAAGGTACCTCAAATCAAGAAACAGAAAAACCAGTTCAACCTGCAGAAGAAAC ACAAACAAACTCTGGGAAAATAGCTAACGAAAATACTGGAGAAGTATCCAATAAACCTAGTGATTCAAA ACCACCAGTTGAAGAATCAAATCAACCAGAAAAAAACGGAACTGCAACAAAACCAGAAAATTCAGGTAA TACAACATCAGAGAATGGACAAACAGAACCAGAACCATCAAACGGAAATTCAACTGAGGATGTTTCAAC CGAATCAAACACATCCAATTCAAATGGAAACGAAGAAATTAAACAAGAAAATGAACTAGACCCTGATAA AAAGGTAGAAGAACCAGAGAAAACACTTGAATTAAGAAATGTTTCCGACCTAGAGTTA
SP004 amino acid (SEQ ID NO : 4 )
NYNTDYELTSGEKLPLPKEISGYTYIGYIKEGKTTSESEVSNQKSSVATPTKQQKVDYNVTPNFVDHPS TVQAIQEQTPVSSTKPTEVQWEKPFSTELINPRKEEKQSSDSQEQLAEHKNLETKKEEKISPKEKTGV NT NPQDEVLSGQLNKPELLYREΞTMETKIDFQEEIQENPDLAEGTVRVKQEGKLGKKVEIVRIFSV K EEVSREIVSTSTTAPSPRIVEKGTKKTQVIKEQPETGVEHKDVQSGAIVEPAIQPELPEAWSDKGΞPE VQPTLPEAWTDKGETEVQPESPDTWSDKGEPEQVAPLPEYKGNIEQVKPETPVEKTKEQGPEKTEEV PVKPTEETPVNPNΞGTTEGTSIQEAENPVQPAEESTTNSEKVSPDTSSK TGEVSSNPSDSTTSVGΞSN KPEHNDΞKNENSEKTVΞEVPVNPNEGTVEGTSNQETEKPVQPAEETQTNSGKIANENTGEVSNKPSDSK PPVEESNQPEK GTATKPENΞGNTTSENGQTEPEPSNGNSTEDVSTESNTSNSNGNEEIKQENELDPDK KVEEPEKTLELRNVSDLEL
SP006 nucleotide (SEQ ID NO : 5 )
TGAGAATCAAGCTACACCCAAAGAGACTAGCGCTCAAAAGACAATCGTCCTTGCTACAGCTGGCGACGT GCCACCATTTGACTACGAAGACAAGGGCAATCTGACAGGCTTTGATATCGAAGTTTTAAAGGCAGTAGA TGAAAAACTCAGCGACTACGAGATTCAATTCCAAAGAACCGCCTGGGAGAGCATCTTCCCAGGACTTGA TTCTGGTCACTATCAGGCTGCGGCCAATAACTTGAGTTACACAAAAGAGCGTGCTGAAAAATACCTTTA CTCGCTTCCAATTTCCAACAATCCCCTCGTCCTTGTCAGCAACAAGAAAAATCCT TGACTTCTCTTGA CCAGATCGCTGGTAAAACAACACAAGAGGATACCGGAACTTCTAACGCTCAATTCATCAATAACTGGAA TCAGAAACACACTGATAATCCCGCTACAATTAATTTTTCTGGTGAGGATATTGGTAAACGAATCCTAGA CCTTGCTAACGGAGAGTTTGATTTCCTAGTTTTTGACAAGGTATCCGTTCAAAAGATTATCAAGGACCG TGGTTTAGACCTCTCAGTCGTTGATTTACCTTCTGCAGATAGCCCCAGCAATTATATCATTTTCTCAAG CGACCAAAAAGAGTTTAAAGAGCAATTTGATAAAGCGCTCAAAGAACTCTATCAAGACGGAACCCTTGA AAAACTCAGCAATACCTATCTAGGTGGTTCTTACCTCCCAGATCAATCTCAGTTACAA
SP006 amino acid (SEQ ID NO: 6)
ENQATPKETSAQKTIVLATAGDVPPFDYEDKGNLTGFDIEVLKAVDEKLSDYEIQFQRTAWESIFPG D SGHYQAAANNLSYTKERAEKYLYSLPISNNPLVLVSNKKNPLTSLDQIAGKTTQEDTGTSNAQFINNWN QKHTDNPATINFSGEDIGKRILDLANGEFDFLVFDKVSVQKIIKDRGLDLSWD PSADSPSNYIIFSS DQKEFKEQFDKALKE YQDGTLEKLSNTYLGGSYLPDQSQLQ
SP007 nucleotide (SEQ ID NO : 7 )
TGGTAACCGCTCTTCTCGTAACGCAGCTTCATCTTCTGATGTGAAGACAAAAGCAGCAATCGTCACTGA TACTGGTGGTGTTGATGACAAATCATTCAACCAATCAGCTTGGGAAGGTTTGCAGGCTTGGGGTAAAGA ACACAATCTTTCAAAAGATAACGGTTTCACTTACTTCCAATCAACAAGTGAAGCTGACTACGCTAACAA CTTGCAACAAGCGGCTGGAAGTTACAACCTAATCTTCGGTGTTGGTTTTGCCCTTAATAATGCAGTTAA AGATGCAGCAAAAGAACACACTGACTTGAACTATGTCTTGATTGATGATGTGATTAAAGACCAAAAGAA TGTTGCGAGCGTAACTTTCGCTGATAATGAGTCAGGTTACCTTGCAGGTGTGGCTGCAGCAAAAACAAC TAAGACAAAACAAGTTGGTTTTGTAGGTGGTATCGAATCTGAAGTTATCTCTCGTTTTGAAGCAGGATT CAAGGCTGGTGTTGCGTCAGTAGACCCATCTATCAAAGTCCAAGTTGACTACGCTGGTTCATTTGGTGA TGCGGCTAAAGGTAAAACAATTGCAGCCGCACAATACGCAGCCGGTGCAGATATTGTTTACCAAGTAGC TGGTGGTACAGGTGCAGGTGTCTTTGCAGAGGCAAAATCTCTCAACGAAAGCCGTCCTGAAAATGAAAA AGTTTGGGTTATCGGTGTTGATCGTGACCAAGAAGCAGAAGGTAAATACACTTCTAAAGATGGCAAAGA ATCAAACTTTGTTCTTGTATCTACTTTGAAACAAGTTGGTACAACTGTAAAAGATATTTCTAACAAGGC AGAAAGAGGAGAATTCCCTGGCGGTCAAGTGATCGTTTACTCATTGAAGGATAAAGGGGTTGACTTGGC AGTAACAAACCTTTCAGAAGAAGGTAAAAAAGCTGTCGAAGATGCAAAAGCTAAAATCCTTGATGGAAG CGTAAAAGTTCCTGAAAAA Table 1 50
SP007 amino acid (SEQ ID NO : 8 )
GNRSSRNAASSSDVKTKAAIVTDTGGVDDKSFNQSAWEGLQAWGKEHN SKDNGFTYFQSTSEADYANN QQAAGΞYNLIFGVGFALNNAVKDAAKEHTDLNYVLIDDVIKDQKNVASVTFADNESGYLAGVAAAKTT KTKQVGFVGGIESEVISRFEAGFKAGVASVDPΞIKVQVDYAGSFGDAAKGKTIAAAQYAAGADIVYQVA GGTGAGVFAEAKSLNESRPENEKV VIGVDRDQEAEGKYTSKDGKESNFV VSTLKQVGTTVKDISNKA ERGEFPGGQVIVYΞLKDKGVDLAVTNLSEEGKKAVEDAKAKILDGSVKVPEK
SP008 nucleotide (SEQ ID NO : 9 )
TGTGGAAATTTGACAGGTAACAGCAAAAAAGCTGCTGATTCAGGTGACAAACCTGTTATCAAAATGTAC CAAATCGGTGACAAACCAGACAACTTGGATGAATTGTTAGCAAATGCCAACAAAATCATTGAAGAAAAA GTTGGTGCCAAATTGGATATCCAATACCTTGGCTGGGGTGACTATGGTAAGAAAATGTCAGTTATCACA TCATCTGGTGAAAACTATGATATTGCCTTTGCAGATAACTATATTGTAAATGCTCAAAAAGGTGCTTAC GCTGACTTGACAGAATTGTACAAAAAAGAAGGTAAAGACCTTTACAAAGCACTTGACCCAGCTTACATC AAGGGTAATACTGTAAATGGTAAGATTTACGCTGTTCCAGTTGCAGCCAACGTTGCATCATCTCAAAAC TTTGCCTTCAACGGAACTCTCCTTGCTAAATATGGTATCGATATTTCAGGTGTTACTTCTTACGAAACT C TGAGCCAGTCTTGAAACAAATCAAAGAAAAAGCTCCAGACGTAGTACCATTTGCTATTGGTAAAGTT TTCATCCCATCTGATAATTTTGACTACCCAGTAGCAAACGGTCTTCCATTCGTTATCGACCTTGAAGGC GATACTACTAAAGTTGTAAACCGTTACGAAGTGCCTCGTTTCAAAGAACACTTGAAGACTCTTCACAAA TTCTATGAAGCTGGCTACATTCCAAAAGACGTCGCAACAAGCGATACTTCCTTTGACCTTCAACAAGAT ACTTGGTTCGTTCGTGAAGAAACAGTAGGACCAGCTGACTACGGTAACAGCTTGCTTTCACGTGTTGCC AACAAAGATATCCAAATCAAACCAATTACTAACTTCATCAAGNAAAACCAAACAACACAAGTTGCTAAC TTTGTCATCTCAAACAACTCTAAGAACAAAGAAAAATCAATGGAAATCTTGAACCTCTTGAATACGAAC CCAGAACTCTTGAACGGTCTTGTTTACGGTCCAGAAGGCAAGAACTGGGAAAAAATTGAAGGTAAAGAA AACCGTGTTCGCGT eTTGATGGCTACAAAGGAAACACTCACATGGGTGGATGGAACACTGGTAACAAC TGGATCCTTTACATCAACGAAAACGTTACAGACCAACAAATCGAAAATTCTAAGAAAGAATTGGCAGAA GCTAAAGAATCTCCAGCGCTTGGATTTATCTTCAATACTGACAATGTGAAATCTGAAATCTCAGCTATT GCTAACACAATGCAACAATTTGATACAGCTATCAACACTGGTACTGTAGACCCAGATAAAGCGATTCCA GAATTGATGGAAAAATTGAAATCTGAAGGTGCCTACGAAAAAGTATTGAACGAAATGCAAAAACAATAC GATGAATTCTTGAAAAACAAAAAA
SP008 amino acid (SEQ ID NO: 10)
CGNLTGNSKKAADSGDKPVIKMYQIGDKPDNLDE LANANKIIEEKVGAKLDIQYLG GDYGKKMSVIT S3GENYDIAFADNYIVNAQKGAYADLTELYKKEGKDLYKALDPAYIKGNTVNGKIYAVPVAANVASSQN FAFNGTLLAKYGIDISGVTSYETLEPVLKQIKEKAPDWPFAIGKVFIPSDNFDYPVANGLPFVIDLΞG DTTKWNRYEVPRFKEHLKTLHKFYEAGYIPKDVATSDTSFDLQQDTWFVREETVGPADYGNSLLSRVA NKDIQIKPITNFIKXNQTTQVANFVISNNSKNKΞKSMEILNLLNTNPELLNGLVYGPEGKN EKIEGKE NRVRV DGYKGNTH GG NTGNN ILYINENVTDQQIENSKKE AEAKESPALGFIFNTDNV ΞEISAI ANTMQQFDTAINTGTVDPDKAIPELMEKLKSEGAYEKVLNEMQKQYDEFLKNKK
SP009 nucleotide (SEQ ID NO: 11)
TGGTCAAGGAACTGCTTCTAAAGACAACAAAGAGGCAGAACTTAAGAAGGTTGACTTTATCCTAGACTG GACACCAAATACCAACCACACAGGGCTTTATGTTGCCAAGGAAAAAGGTTATTTCAAAGAAGCTGGAGT GGATGTTGATTTGAAATTGCCACCAGAAGAAAGTTCTTCTGACTTGGTTATCAACGGAAAGGCACCATT TGCAGTGTATTTCCAAGACTACATGGCTAAGAAATTGGAAAAAGGAGCAGGAATCACTGCCGTTGCAGC TATTGTTGAACACAATACATCAGGAATCATCTCTCGTAAATCTGATAATGTAAGCAGTCCAAAAGACTT GGTTGGTAAGAAATATGGGACATGGAATGACCCAACTGAACTTGCTATGTTGAAAACCTTGGTAGAATC TCAAGGTGGAGACTTTGAGAAGGTTGAAAAAGTACCAAATAACGACTCAAACTCAATCACACCGATTGC CAATGGCGTCTTTGATACTGCTTGGATTTACTACGGTTGGGATGGTATCCTTGCTAAATCTCAAGGTGT AGATGCTAACTTCATGTACTTGAAAGACTATGTCAAGGAGTTTGACTACTATTCACCAGTTATCATCGC AAACAACGACTATCTGAAAGATAACAAAGAAGAAGCTCGCAAAGTCATCCAAGCCATCAAAAAAGGCTA CCAATATGCCATGGAACATCCAGAAGAAGCTGCAGATATTCTCATCAAGAATGCACCTGAACTCAAGGA AAAACGTGACTTTGTCATCGAATCTCAAAAATACTTGTCAAAAGAATACGCAAGCGACAAGGAAAAATG GGGTCAATTTGACGCAGCTCGCTGGAATGCTTTCTACAAATGGGATAAAGAAAATGGTATCCTTAAAGA AGACTTGACAGACAAAGGCTTCACCAACGAATTTGTGAAA
SP009 amino acid (SEQ ID NO:12) Table 1 51
GQGTASKDNKEAELKKVDFILDWTPNTNHTG YVAKEKGYFKEAGVDVDLKLPPEESSSDLVINGKAPF AVYFQDYMAKKLEKGAGITAVAAIVEHNTSGIISRKSDNVSSPKDLVGKKYGT DPTELA LKTLVES QGGDFEKVEKVP NDSNSITPIANGVFDTAWIYYGWDGILAKSQGVDANFMYLKDYVKEFDYYSPVIIA NNDYLKDNKEEARKVIQAIKKGYQYAMEHPEEAADILIKNAPELKEKRDFVIESQKY SKEYASDKEK GQFDAARWNAFYK DKENGILKEDLTDKGFTNEFVK
SP010 nucleotide (SEQ ID NO: 13)
TAGCTCAGGTGGAAACGCTGGTTCATCCTCTGGAAAAACAACTGCCAAAGCTCGCACTATCGATGAAAT CAAAAAAAGCGGTGAACTGCGAATCGCCGTGTTTGGAGATAAAAAACCGTTTGGCTACGTTGACAATGA TGGTTCTACCAAGGTACGCTACGATATTGAACTAGGGAACCAACTAGCTCAAGACCTTGGTGTCAAGGT TAAATACATTTCAGTCGATGCTGCCAACCGTGCGGAATACTTGATTTCAAACAAGGTAGATATTACTCT TGCTAACTTTACAGTAACTGACGAACGTAAGAAACAAGTTGATTTTGCCCTTCCATATATGAAAGTTTC TCTGGGTGTCGTATCACCTAAGACTGGTCTCATTACAGACGTCAAACAACTTGAAGGTAAAACCTTAAT TGTCACAAAAGGAACGACTGCTGAGACTTATTTTGAAAAGAATCATCCAGAAATCAAACTCCAAAAATA CGACCAATACAGTGACTCTTACCAAGCTCTTCTTGACGGACGTGGAGATGCCTTTTCAACTGACAATAC GGAAGTTCTAGCTTGGGCGCTTGAAAATAAAGGATTTGAAGTAGGAATTACTTCCCTCGGTGATCCCGA TACCATTGCGGCAGCAGTTCAAAAAGGCAACCAAGAATTGCTAGACTTCATCAATAAAGATATTGAAAA ATTAGGCAAGGAAAACTTCTTCCACAAGGCCTATGAAAAGACACTTCACCCAACCTACGGTGACGCTGC TAAAGCAGATGACCTGGTTGTTGAAGGTGGAAAAGTTGAT
SP010 amino acid (SEQ ID NO:14)
SSGGNAGSSSGKTTAKARTIDEIKKSGELRIAVFGDKKPFGYVDNDGSTKVRYDIELGNQ AQDLGVKV KYISVOAANRAEYLISNKVDITLANFTVTDERKKQVDFALPYMKVSLGVVSPKTGLITDVKQLEGKTLI VTKGTTAETYFE NHPEIKLQKYDQYSDSYQALLDGRGDAFSTDNTEVLA ALENKGFEVGITSLGDPD TIAAAVQKGNQELLDFINKDIEKLGKENFFHKAYΞKTLHPTYGDAAKADDLWEGGKVD
SP011 nucleotide (SEQ ID NO: 15)
CTCCAACTATGGTAAATCTGCGGATGGCACAGTGACCATCGAGTATTTCAACCAGAAAAAAGAAATGAC CAAAACCTTGGAAGAAATCACTCGTGATTTTGAGAAGGAAAACCCTAAGATCAAGGTCAAAGTCGTCAA TGTACCAAATGCTGGTGAAGTATTGAAGACACGCGTTCTCGCAGGAGATGTGCCTGATGTGGTCAATAT TTACCCACAGTCCATCGAACTGCAAGAATGGGCAAAAGCAGGTGTTTTTGAAGATTTGAGCAACAAAGA CTACCTGAAACGCGTGAAAAATGGCTACGCTGAAAAATATGCTGTAAACGAAAAAGTTTACAACGTTCC TTTTACAGCTAATGCTTATGGAATTTACTACAACAAAGATAAATTCGAAGAACTGGGCTTGAAGGTTCC TGAAACCTGGGATGAATTTGAACAGTTAGTCAAAGATATCGTTGCTAAAGGACAAACACCATTTGGAAT TGCAGGTGCAGATGCTTGGACACTCAATGGTTACAATCAATTAGCCTTTGCGACAGCAACAGGTGGAGG AAAAGAAGCAAATCAATACCTTCGTTATTCTCAACCAAATGCCATTAAATTGTCGGATCCGATTATGAA AGATGATATCAAGGTCATGGACATCCTTCGCATCAATGGATCTAAGCAAAAGAACTGGGAAGGTGCTGG CTATACCGATGTTATCGGAGCCTTCGCACGTGGGGATGTCCTCATGACACCAAATGGGTCTTGGGCGAT CACAGCGATTAATGAACAAAAACCGAACTTTAAGATTGGGACCTTCATGATTCCAGGAAAAGAAAAAGG ACAAAGCTTAACCGTTGGTGCGGGAGACTTGGCATGGTCTATCTCAGCCACCACCAAACATCCAAAAGA AGCCAATGCCTTTGTGGAATATATGACCCGTCCAGAAGTCATGCAAAAATACTACGATGTGGACGGATC TCCAACAGCGATCGAAGGGGTCAAACAAGCAGGAGAAGATTCACCGCTTGCTGGTATGACCGAATATGC CTTTACGGATCGTCACTTGGTCTGGTTGCAACAATACTGGACCAGTGAAGCAGACTTCCATACCTTGAC CATGAACTATGTCTTGACCGGTGATAAACAAGGCATGGTCAATGATTTGAATGCCTTCTTTAACCCGAT GAAAGCGGATGTGGAT
SP011 amino acid (SEQ ID NO: 16)
SNYGKΞADGTVTIEYFNQKKEMTKTLEEITRDFEKENPKIKV1 A/NVPNAGEVLKTRVLAGDVPDVVNI YPQSIΞLQEWAKAGVFEDLSNKDY KRVKNGYAEKYAVNEKVYNVPFTANAYGIYY KDKFEELGLKVP ET DEFEQLV DIVAKGQTPFGIAGADAWTLNGYNQLAFATATGGGKEANQYLRYSQPNAIKLSDPIMK DDIKVMDILRINGSKQ NWEGAGYTDVIGAFARGDVLMTPNGS AITAINEQKPNFKIGTF IPGKEKG QSLTVGAGDIΛWSISATTKΗPKE-ANAFVEYMTRPEVMQKYYDVDGSPTAIEGVKQAGEDSPLAGMTEYA FTDRH VWLQQYVJTSEADFKTLTMNYVLTGDKQGMV D NAFFNPMKADVD
SP012 nucleotide (SEQ ID NO:17)
TGGGAAAAATTCTAGCGAAACTAGTGGAGATAATTGGTCAAAGTACCAGTCTAACAAGTCTATTACTAT TGGATTTGATAGTACTTTTGTTCCAATGGGATTTGCTCAGAAAGATGGTTCTTATGCAGGATTTGATAT TGATTTAGCTACAGCTGTTTTTGAAAAATACGGAATCACGGTAAATTGGCAACCGATTGATTGGGATTT Table 1 52
GAAAGAAGCTGAATTGACAAAAGGAACGATTGATCTGATTTGGAATGGCTATTCCGCTACAGACGAACG CCGTGAAAAGGTGGCTTTCAGTAACTCATATATGAAGAATGAGCAGGTATTGGTTACGAAGAAATCATC TGGTATCACGACTGCAAAGGATATGACTGGAAAGACATTAGGAGCTCAAGCTGGTTCATCTGGTTATGC GGACTTTGAAGCAAATCCAGAAATTTTGAAGAATATTGTCGCTAATAAGGAAGCGAATCAATACCAAAC CTTTAATGAAGCCTTGATTGATTTGAAAAACGATCGAATTGATGGTCTATTGATTGACCGTGTCTATGC AAACTATTATTTAGAAGCAGAAGGTGTTTTAAACGATTATAATGTCTTTACAGTTGGACTAGAAACAGA AGCTTTTGCGGTTGGAGCCCGTAAGGAAGATACAAACTTGGTTAAGAAGATAAATGAAGCTTTTTCTAG TCTTTACAAGGACGGCAAGTTCCAAGAAATCAGCCAAAAATGGTTTGGAGAAGATGTAGCAACCAAAGA AGTAAAAGAAGGACAG
SP012 nucleotide (SEQ ID NO: 18)
GK SSETSGD SKYQSNKSITIGFDSTFVPMGFAQKDGSYAGFDIDLATAVFEKYGITVNWQPIDWDL KEAELTKGTIDLI NGYSATDERREKVAFSNSYMKNEQVLVTKKSSGITTAKDMTGKTLGAQAGSSGYA DFEANPEILKMIVANKEA QYQTFNEALIDLKNDRIDGL IDRVΥANYYLEAEGVLNDYNVFTVG ETE AFAVGARKEDTNLVKKINEAFSSLYKDGKFQEISQKWFGEDVATKEVKEGQ
SP013 nucleotide (SEQ ID NO:19)
TGCTAGCGGAAAAAAAGATACAACTTCTGGTCAAAAACTAAAAGTTGTTGCTACAAACTCAATCATCGC TGATATTACTAAAAATATTGCTGGTGACAAAATTGACCTTCATAGTATCGTTCCGATTGGGCAAGACCC ACACGAATACGAACCACTTCCTGAAGACGTTAAGAAAACTTCTGAGGCTAATTTGATTTTCTATAACGG TATCAACCTTGAAACAGGTGGCAATGCTTGGTTTACAAAATTGGTAGAAAATGCCAAGAAAACTGAAAA CAAAGACTACTTCGCAGTCAGCGACGGCGTTGATGTTATCTACCTTGAAGGTCAAAATGAAAAAGGAAA AGAAGACCCACACGCTTGGCTTAACCTTGAAAACGGTATTATTTTTGCTAAAAATATCGCCAAACAATT GAGCGCCAAAGACCCTAACAATAAAGAATTCTATGAAAAAAATCTCAAAGAATATACTGATAAGTTAGA CAAACTTGATAAAGAAAGTAAGGATAAATTTAATAAGATCCCTGCTGAAAAGAAACTCATTGTAACCAG CGAAGGAGCATTCAAATACTTCTCTAAAGCCTATGGTGTCCCAAGTGCTTACATCTGGGAAATCAATAC TGAAGAAGAAGGAACTCCTGAACAAATCAAGACCTTGGTTGAAAAACTTCGCCAAACAAAAGTTCCATC ACTCTTTGTAGAATCAAGTGTGGATGACCGTCCAATGAAAACTGTTTCTCAAGACACAAACATCCCAAT CTACGCTCAAATCTTTACTGACTCTATCGCAGAACAAGGTAAAGAAGGCGACAGCTACTACAGCATGAT GAAATACAACCTTGACAAGATTGCTGAAGGATTGGCAAAA
SP013 amino acid (SEQ ID NO:20)
ASGKKDTTΞGQK KWATNSIIADITKNIAGDKIDLHΞIVPIGQDPHEYEPLPEDVKKTΞEANLIFYNG INLETGGNAWFTKLVENAKKTENKDYFAVSDGVDVIYLEGQNEKGKΞDPHAWLN ENGIIFAKNIAKQL SAKDPNNKEFYE MLKEYTDKLDKLDKESKDKFNKIPAEKKLIVTSEGAFKYFSKAYGVPSAYIWEINT ΞΞEGTPEQIKTLVEKLRQTKVPSLFVESSVDDRPMKTVSQDTNIPIYAQIFTDΞIAEQGKEGDSYYSMM KYNLDKIAEGLAK
SP014 nucleotide (SEQ ID NO: 21)
TGGCTCAAAAAATACAGCTTCAAGTCCAGATTATAAGTTGGAAGGTGTAACATTCCCGCTTCAAGAAAA GAAAACATTGAAGTTTATGACAGCCAGTTCACCGTTATCTCCTAAAGACCCAAATGAAAAGTTAATTTT GCAACGTTTGGAGAAGGAAACTGGCGTTCATATTGACTGGACCAACTACCAATCCGACTTTGCAGAAAA ACGTAACTTGGATATTTCTAGTGGTGATTTACCAGATGCTATCCACAACGACGGAGCTTCAGATGTGGA CTTGATGAACTGGGCTAAAAAAGGTGTTATTATTCCAGTTGAAGATTTGATTGATAAATACATGCCAAA TCTTAAGAAAATTTTGGATGAGAAACCAGAGTACAAGGCCTTGATGACAGCACCTGATGGGCACATTTA CTCATTTCCATGGATTGAAGAGCTTGGAGATGGTAAAGAGTCTATTCACAGTGTCAACGATATGGCTTG GATTAACAAAGATTGGCTTAAGAAACTTGGTCTTGAAATGCCAAAAACTACTGATGATTTGATTAAAGT CCTAGAAGCTTTCAAAAACGGGGATCCAAATGGAAATGGAGAGGCTGATGAAATTCCATTTTCATTTAT TAGTGGTAACGGAAACGAAGATTTTAAATTCCTATTTGCTGCATTTGGTATAGGGGATAACGATGATCA TTTAGTAGTAGGAAATGATGGCAAAGTTGACTTCACAGCAGATAACGATAACTATAAAGAAGGTGTCAA ATTTATCCGTCAATTGCAAGAAAAAGGCCTGATTGATAAAGAAGCTTTCGAACATGATTGGAATAGTTA CATTGCTAAAGGTCATGATCAGAAATTTGGTGTTTACTTTACATGGGATAAGAATAATGTTACTGGAAG TAACGAAAGTTATGATGTTTTACCAGTACTTGCTGGACCAAGTGGTCAAAAACACGTAGCTCGTACAAA CGGTATGGGATTTGCACGTGACAAGATGGTTATTACCAGTGTAAACAAAAACCTAGAATTGACAGCTAA ATGGATTGATGCACAATACGCTCCACTCCAATCTGTGCAAAATAACTGGGGAACTTACGGAGATGACAA ACAACAAAACATCTTTGAATTGGATCAAGCGTCAAATAGTCTAAAACACTTACCACTAAACGGAACTGC ACCAGCAGAACTTCGTCAAAAGACTGAAGTAGGAGGACCACTAGCTATCCTAGATTCATACTATGGTAA AGTAACAACCATGCCTGATGATGCCAAATGGCGTTTGGATCTTATCAAAGAATATTATGTTCCTTACAT Table 1 53
GAGCAATGTCAATAACTATCCAAGAGTCTTTATGACACAGGAAGATTTGGACAAGATTGCCCATATCGA AGCAGATATGAATGACTATATCTACCGTAAACGTGCTGAATGGATTGTAAATGGCAATATTGATACTGA GTGGGATGATTACAAGAAAGAACTTGAAAAATACGGACTTTCTGATTACCTCGCTATTAAACAAAAATA CTACGACCAATACCAAGCAAACAAAAAC
SP014 amino acid (SEQ ID NO:22)
GSKNTASSPDYKLEGVTFPLQEKKTLKFMTASSPLSPKDPNEKLILQRLEKETGVHID TNYQSDFAEK R LDISSGDLPDAIH DGASDVDLMN AKKGVIIPVEDLIDKYMPNLKKILDEKPEYKALMTAPDGHIY SFP IEELGDGKESIHSVNDMAWINKDW KKLGLEMPKTTDDLIK LEAFKNGDPNGNGEADEIPFSFI SGNGNEDFKFLFAAFGIGDNDDHLWGNDGKVDFTADNDNYKEGλKFIRQLQEKGLIDKEAFEHDWNSY IAKGHDQKFGVYFT DKN VTGSNESYDVLPVLAGPSGQKHVARTNGMGFARDKMVITSVNKNLELTAK WIDAQYAPLQSVQN GTYGDDKQQNIFELDQASNSLKHLPLNGTAPAELRQKTEVGGPLAILDSYYGK VTTMPDDAK RLDLIKEYYVPYMSNVNNYPRVFMTQEDLDKIAHIEAD NDYIYRKRAEWIVNGNIDTE DDYKKELEKYGLSDYLAIKQKYYDQYQA KN
SP015 nucleotide (SEQ ID NO: 23)
TAGTACAAACTCAAGCACTAGTCAGACAGAGACCAGTAGCTCTGCTCCAACAGAGGTAACCATTAAAAG TTCACTGGACGAGGTCAAACTTTCCAAAGTTCCTGAAAAGATTGTGACCTTTGACCTCGGCGCTGCGGA TACTATTCGCGCTTTAGGATTTGAAAAAAATATCGTCGGAATGCCTACAAAAACTGTTCCGACTTATCT AAAAGACCTAGTGGGAACTGTCAAAAATGTTGGTTCTATGAAAGAACCTGATTTAGAAGCTATCGCCGC CCTTGAGCCTGATTTGATTATCGCTTCGCCACGTACACAAAAATTCGTAGACAAATTCAAAGAAATCGC CCCAACCGTTCTCTTCCAAGCAAGCAAGGACGACTACTGGACTTCTACCAAGGCTAATATCGAATCCTT AGCAAGTGCCTTCGGCGAAACTGGTACACAGAAAGCCAAGGAAGAATTGACCAAGCTAGACAAGAGCAT CCAAGAAGTCGCTACTAAAAATGAAAGCTCTGACAAAAAAGCCCTTGCGATCCTCCTTAATGAAGGAAA AATGGCAGCCTTTGGTGCCAAATCTCGTTTCTCTTTCTTGTACCAAACCTTGAAATTCAAACCAACTGA TACAAAATTTGAAGACTCACGCCACGGACAAGAAGTCAGCTTTGAAAGTGTCAAAGAAATCAACCCTGA CATCCTCTTTGTCATCAACCGTACCCTTGCCATCGGTGGGGACAACTCTAGCAACGACGGTGTCCTAGA AAATGCCCTTATCGCTGAAACACCTGCTGCTAAAAATGGTAAGATTATCCAACTAACACCAGACCTCTG GTATCTAAGCGGAGGCGGACTTGAATCAACAAAACTCATGATTGAAGACATACAAAAAGCTTTGAAA
SP015 amino acid (SEQ ID NO: 24)
STNSSTSQTETSSSAPTEVTIKSΞLDEVKLSKVPEKIVTFDLGAADTIRALGFEKNIVG PTKTVPTYL KDLVGTVKNVGSMKEPDLEAIAALEPDLIIASPRTQKFVDKFKEIAPTVLFQASKDDYWTSTKANIESL ASAFGETGTQKAKEELTKLDKSIQEVAT NESSDKKALAILLNΞGKMAAFGAK5RFΞFLYQTLKFKPTD TKFEDSRHGQEVSFESVKEINPDILFVINRTLAIGGDNSSNDGVLENA IAETPAAKNGKIIQLTPDL YLSGGGLESTKLMIEDIQKALK
SP016 nucleotide (SEQ ID NO:25)
TGGCAATTCTGGCGGAAGTAAAGATGCTGCCAAATCAGGTGGTGACGGTGCCAAAACAGAAATCACTTG GTGGGCATTCCCAGTATTTACCCAAGAAAAAACTGGTGACGGTGTTGGAACTTATGAAAAATCAATCAT CGAAGCGTTTGAAAAAGCAAACCCAGATATAAAAGTGAAATTGGAAACCATCGACTTCAAGTCAGGTCC TGAAAAAATCACAACAGCCATCGAAGCAGGAACAGCTCCAGACGTACTCTTTGATGCACCAGGACGTAT CATCCAATACGGTAAAAACGGTAAATTGGCTGAGTTGAATGACCTCTTCACAGATGAATTTGTTAAAGA TGTCAACAATGAAAACATCGTACAAGCAAGTAAAGCTGGAGACAAGGCTTATATGTATCCGATTAGTTC TGCCCCATTCTACATGGCAATGAACAAGAAAATGTTAGAAGATGCTGGAGTAGCAAACCTTGTAAAAGA AGGTTGGACAACTGATGATTTTGAAAAAGTATTGAAAGCACTTAAAGACAAGGGTTACACACCAGGTTC ATTGTTCAGTTCTGGTCAAGGGGGAGACCAAGGAACACGTGCCTTTATCTCTAACCTTTATAGCGGTTC TGTAACAGATGAAAAAGTTAGCAAATATACAACTGATGATCCTAAATTCGTCAAAGGTCTTGAAAAAGC AACTAGCTGGATTAAAGACAATTTGATCAATAATGGTTCACAATTTGACGGTGGGGCAGATATCCAAAA CTTTGCCAACGGTCAAACATCTTACACAATCCTTTGGGCACCAGCTCAAAATGGTATCCAAGCTAAACT TTTAGAAGCAAGTAAGGTAGAAGTGGTAGAAGTACCATTCCCATCAGACGAAGGTAAGCCAGCTCTTGA GTACCTTGTAAACGGGTTTGCAGTATTCAACAATAAAGACGACAAGAAAGTCGCTGCATCTAAGAAATT CATCCAGTTTATCGCAGATGACAAGGAGTGGGGACCTAAAGACGTAGTTCGTACAGGTGCTTTCCCAGT CCGTACTTCATTTGGAAAACTTTATGAAGACAAACGCATGGAAACAATCAGCGGCTGGACTCAATACTA CTCACCATACTACAACACTATTGATGGATTTGCTGAAATGAGAACACTTTGGTTCCCAATGTTGCAATC TGTATCAAATGGTGACGAAAAACCAGCAGATGCTTTGAAAGCCTTCACTGAAAAAGCGAACGAAACAAT CAAAAAAGCTATGAAACAA Table 1 54
SP016 amino acid (SEQ ID NO:26)
GNSGGΞKDAAKSGGDGAKTEITW AFPVFTQEKTGDGVGTYEKSIIEAFEKANPDIKVKLETIDFKSGP EKITTAIEAGTAPDVLFDAPGRIIQYGKNGKLAELNDLFTDEFVKDVNNENIVQASKAGDKAYMYPISS APFYMAMNKKMLEDAGVA LVKEG TTDDFEKVLKALKDKGYTPGΞLFSSGQGGDQGTRAFISNLYΞGS VTDEKVSKYTTDDPKFVKGLEKATS IKDN INNGSQFDGGADIQNFANGQTSYTIL APAQNGIQAKL LF-ASKVFΛ^VEVPFPSDEGKPALEYLVNGFAVF NKDDKKVAASKKFIQFIADDKE GPKDVVRTGAFPV RTSFGKLYEDKRMETISG TQYYSPYYNTIDGFAEMRTLWFP LQSVΞNGDEKPADALKAFTEKANETI KKAMKQ
SP017 nucleotide (SEQ ID NO:27)
TTCACAAGAAAAAACAAAAAATGAAGATGGAGAAACTAAGACAGAACAGACAGCCAAAGCTGATGGAAC AGTCGGTAGTAAGTCTCAAGGAGCTGCCCAGAAGAAAGCAGAAGTGGTCAATAAAGGTGATTACTACAG CATTCAAGGGAAATACGATGAAATCATCGTAGCCAACAAACACTATCCATTGTCTAAAGACTATAATCC AGGGGAAAATCCAACAGCCAAGGCAGAGTTGGTCAAACTCATCAAAGCGATGCAAGAGGCAGGTTTCCC TATTAGTGATCATTACAGTGGTTTTAGAAGTTATGAAACTCAGACCAAGCTCTATCAAGATTATGTCAA CCAAGATGGAAAGGCAGCAGCTGACCGTTACTCTGCCCGTCCTGGCTATAGCGAACACCAGACAGGCTT GGCCTTTGATGTGATTGGGACTGATGGTGATTTGGTGACAGAAGAAAAAGCAGCCCAATGGCTCTTGGA TCATGCAGCTGATTATGGCTTTGTTGTCCGTTATCTCAAAGGCAAGGAAAAGGAAACAGGCTATATGGC TGAAGAATGGCACCTGCGTTATGTAGGAAAAGAAGCTAAAGAAATTGCTGCAAGTGGTCTCAGTTTGGA AGAATACTATGGCTTTGAAGGCGGAGACTACGTCGAT
SP017 amino acid (SEQ ID NO:28)
SQEKTKNEDGETKTEQTAKADGTVGSKSQGAAQKKAEλA/NKGDYYSIQGKYDEIIVANKHYPLSKDYNP GENPTAKAELVKLIKAMQEAGFPISDHYSGFRSYΞTQTKLYQDYVNQDGKAAADRYSARPGYSEHQTGL AFDVIGTDGDLVTΞEKAAQ LLDHAADYGFVVRYLKGKEKETGYMAEE HLRYVGKEAKEIAASGLSLE ΞYYGFEGGDYVD
SP019 nucleotide (SEQ ID NO:29)
GAAAGGTCTGTGGTCAAATAATCTTACCTGCGGTTATGATGAAAAAATAATCTTGGAAAATATAAATAT AAAAATACCTGAAGAAAAAATATCAGTTATTATTGGGTCAAATGGTTGTGGGAAATCAACACTCATTAA AACCTTGTCTCGACTTATAAAGCCATTAGAGGGAGAAGTATTGCTTGATAATAAATCAATTAATTCTTA TAAAGAAAAAGATTTAGCAAAACACATAGCTATATTACCTCAATCTCCAATAATCCCTGAATCAATAAC AGTAGCTGATCTTGTAAGCCGTGGTCGTTTCCCCTACAGAAAGCCTTTTAAGAGTCTTGGAAAAGATGA CCTTGAAATAATAAACAGATCAATGGTTAAGGCCAATGTTGAAGATCTAGCAAATAACCTAGTTGAAGA ACTTTCTGGGGGTCAAAGGCAAAGAGTATGGATAGCTCTAGCCCTAGCCCAAGATACAAGTATCCTACT TTTAGATGAGCCAACTACTTACTTGGATATCTCATATCAAATAGAACTATTAGACCTCTTGACTGATCT AAACCAAAAATATAAGACAACCATTTGCATGATTTTGCACGATATAAATCTAACAGCAAGATACGCTGA TTACCTATTTGCAATTAAAGAAGGTAAACTTGTTGCAGAGGGAAAGCCTGAAGATATACTAAATGATAA ACTAGTTAAAGATATCTTTAATCTTGAAGCAAAAATTATACGTGACCCTATTTCCAATTCGCCTCTAAT GATTCCTATTGGCAAGCACCATGTTAACTCT
SP019 amino acid (SEQ ID NO:30)
KGL S N TCGYDEKIILENINIKIPEEKISVIIGSNGCGKSTLIKT SRLIKPLEGEVLLDNKSINΞY KEKDLAKHIAILPQSPIIPESITVADLVSRGRFPYRKPFKSLGKDDLEIINRSMVKAVEDLANNLVEE LSGGQRQRV IALALAQDTSI LLDEPTTYLDISYQIELLDLLTDLNQKYKTTICMILHDINLTARYAD YLFAIKEGKLVAEGKPEDILNDKLVKDIFNLEAKIIRDPISNSPLMIPIGKHHVS
SP020 nucleotide (SEQ ID NO:31)
AAACTCAGAAAAGAAAGCAGACAATGCAACAACTATCAAAATCGCAACTGTTAACCGTAGCGGTTCTGA AGAAAAACGTTGGGACAAAATCCAAGAATTGGTTAAAAAAGACGGAATTACCTTGGAATTTACAGAGTT CACAGACTACTCACAACCAAACAAAGCAACTGCTGATGGCGAAGTAGATTTGAACGCTTTCCAACACTA TAACTTCTTGAACAACTGGAACAAAGAAAACGGAAAAGACCTTGTAGCGATTGCAGATACTTACATCTC TCCAATCCGCCTTTACTCAGGTTTGAATGGAAGTGCCAACAAGTACACTAAAGTAGAAGACATCCCAGC AAACGGAGAAATCGCTGTACCGAATGACGCTACAAACGAAAGCCGTGCGCTTTATTTGCTTCAATCAGC TGGCTTGATTAAATTGGATGTTTCTGGAACTGCTCTTGCAACAGTTGCCAACATCAAAGAAAATCCAAA GAACTTGAAAATCACTGAATTGGACGCTAGCCAAACAGCTCGTTCATTGTCATCAGTTGACGCTGCCGT TGTAAACAATACCTTCGTTACAGAAGCAAAATTGGACTACAAGAAATCACTTTTCAAAGAACAAGCTGA TGAAAACTCAAAACAATGGTACAACATCATTGTTGCAAAAAAAGATTGGGAAACATCACCTAAGGCTGA Table 1 55
TGCTATCAAGAAAGTAATCGCAGCTTACCACACAGATGACGTGAAAAAAGTTATCGAAGAATCATCAGA TGGTTTGGATCAACCAGTTTGG
SP020 amino acid (SEQ ID NO:32)
NSEKKADNATTIKIATVNRSGSEEKR DKIQELVKKDGITLEFTEFTDYSQPNKATADGEVDLNAFQHY NFLNN NKENGKDLVAIADTYISPIRLYSG NGSANKYTKVEDIPANGEIAVPNDATNESRALYLLQSA GLIKLDVSGTAI-ATVANIKENPK LKITELDASQTARSLSSVDAAVVNNTFVTEAKLDYKKSLFKEQAD ENSKQWYNIIVAKKD ETSPKADAIKKVIAAYHTDDVKKVIEESΞDGLDQP'VW
SP021 nucleotide (SEQ ID NO:33)
TTCGAAAGGGTCAGAAGGTGCAGACCTTATCAGCATGAAAGGGGATGTCATTACAGAACATCAATTTTA TGAGCAAGTGAAAAGCAACCCTTCAGCCCAACAAGTCTTGTTAAATATGACCATCCAAAAAGTTTTTGA AAAACAATATGGCTCAGAGCTTGATGATAAAGAGGTTGATGATACTATTGCCGAAGAAAAAAAACAATA TGGCGAAAACTACCAACGTGTCTTGTCACAAGCAGGTATGACTCTTGAAACACGTAAAGCTCAAATTCG TACAAGTAAATTAGTTGAGTTGGCAGTTAAGAAGGTAGCAGAAGCTGAATTGACAGATGAAGCCTATAA GAAAGCCTTTGATGAGTACACTCCAGATGTAACGGCTCAAATCATCCGTCTTAATAATGAAGATAAGGC CAAAGAAGTTCTCGAAAAAGCCAAGGCAGAAGGTGCTGATTTTGCTCAATTAGCCAAAGATAATTCAAC TGATGAAAAAACAAAAGAAAATGGTGGAGAAATTACCTTTGATTCTGCTTCAACAGAAGTACCTGGAGC AAGTCCAAAAAAGCCGCTTTTCGCTTTTAGATGTGGGATGGTGTTTCTGGATGTGGATTACAGCAACTG GGGCACACCAAGCCTACAG
SP021 amino acid (SEQ ID NO:34)
SKGSEGADLISMKGDVITEHQFYEQV SNPSAQQVLLNMTIQKVFEKQYGSELDDKEVDDTIAEEKKQY GENYQRVLSQAGMTLETRKAQIRTSKLVELAVKKVAEAELTDEAYKKAFDEYTPDVTAQIIRLNNEDKA KEV EKAKAEGADFAQLAKDNSTDEKTKENGGEITFDSASTEVPGASPKKP FAFRCGMVFLDVDYSNW GTPSLQ
SP022 nucleotide (SEQ ID NO:35)
GGGGATGGCAGCTTTTAAAAATCCTAACAATCAATACAAAGCTATTACAATTGCTCAAACTCTAGGTGA TGATGCTTCTTCAGAGGAATTGGCTGGTAGATATGGTTCTGCTGTTCAGTGTACAGAAGTGACTGCCTC AAACCTTTCAACAGTTAAAACTAAAGCTACGGTTGTAGAAAAACCACTGAAAGATTTTAGAGCGTCTAC GTCTGATCAGTCTGGTTGGGTGGAATCTAATGGTAAATGGTATTTCTATGAGTCTGGTGATGTGAAGAC AGGTTGGGTGAAAACAGATGGTAAATGGTACTATTTGAATGACTTAGGTGTCATGCAGACTGGATTTGT AAAATTTTCTGGTAGCTGGTATTACTTGAGCAATTCAGGTGCTATGTTTACAGGCTGGGGAACAGATGG TAGCAGATGGTTCTACTTTGACGGCTCAGGAGCTATGAAGACAGGCTGGTACAAGGAAAATGGCACTTG GTATTACCTTGACGAAGCAGGTATCATGAAGACAGGTTGGTTTAAAGTCGGACCACACTGGTACTATGC CTACGGTTCAGGAGCTTTGGCTGTGAGCACAACAACACCAGATGGTTACCGTGTAAATGGTAATGGTGA ATGGGTAAAC
SP022 amino acid (SEQ ID NO:36)
GMAAFKNPNNQYKAITIAQT GDDASSEELAGRYGSAVQCTEVTASNLSTVKTKATWEKP KDFRAST SDQSG VESNGKWYFYEΞGDVKTG VKTDGKWYYLNDLGVMQTGFVKFSGS YY SNSGAMFTG GTDG SR FYFDGSGAMKTGWYKENGT YYLDEAGIMKTG FKVGPH YYAYGSGALAVSTTTPDGYRVNGNGE WVN
SP023 nucleotide (SEQ ID NO:37)
AGACGAGCAAAAAATTAAGCAAGCAGAAGCGGAAGTTGAGAGTAAACAAGCTGAGGCTACAAGGTTAAA AAAAATCAAGACAGATCGTGAAGAAGCAGAAGAAGAAGCTAAACGAAGAGCAGATGCTAAAGAGCAAGG TAAACCAAAGGGGCGGGCAAAACGAGGAGTTCCTGGAGAGCTAGCAACACCTGATAAAAAAGAAAATGA TGCGAAGTCTTCAGATTCTAGCGTAGGTGAAGAAACTCTTCCAAGCCCATCCCTGAAACCAGAAAAAAA GGTAGCAGAAGCTGAGAAGAAGGTTGAAGAAGCTAAGAAAAAAGCCGAGGATCAAAAAGAAGAAGATCG CCGTAACTACCCAACCAATACTTACAAAACGCTTGAACTTGAAATTGCTGAGTCCGATGTGGAAGTTAA AAAAGCGGAGCTTGAACTAGTAAAAGAGGAAGCTAAGGAACCTCGAAACGAGGAAAAAGTTAAGCAAGC AAAAGCGGAAGTTGAGAGTAAAAAAGCTGAGGCTACAAGGTTAGAAAAAATCAAGACAGATCGTAAAAA AGCAGAAGAAGAAGCTAAACGAAAAGCAGCAGAAGAAGATAAAGTTAAAGAAAAACCAGCTGAACAACC ACAACCAGCGCCGGCTCCAAAAGCAGAAAAACCAGCTCCAGCTCCAAAACCAGAGAATCCAGCTGAACA ACCAAAAGCAGAAAAACCAGCTGATCAACAAGCTGAAGAAGACTATGCTCGTAGATCAGAAGAAGAATA TAATCGCTTGACTCAACAGCAACCGCCAAAAACTGAAAAACCAGCACAACCATCTACTCCAAAAACAGG Table 1 56
CTGGAAACAAGAAAACGGTATGTGGTACTTCTACAATACTGATGGTTCAATGGCGACAGGATGGCTCCA AAACAATGGCTCATGGTACTACCTCAACAGCAATGGCGCTATGGCGACAGGATGGCTCCAAAACAATGG TTCATGGTACTATCTAAACGCTAATGGTTCAATGGCAACAGGATGGCTCCAAAACAATGGTTCATGGTA CTACCTAAACGCTAATGGTTCAATGGCGACAGGATGGCTCCAATACAATGGCTCATGGTACTACCTAAA CGCTAATGGTTCAATGGCGACAGGATGGCTCCAATACAATGGCTCATGGTACTACCTAAACGCTAATGG TGATATGGCGACAGGTTGGGTGAAAGATGGAGATACCTGGTACTATCTTGAAGCATCAGGTGCTATGAA AGCAAGCCAATGGTTCAAAGTATCAGATAAATGGTACTATGTCAATGGCTCAGGTGCCCTTGCAGTCAA CACAACTGTAGATGGCTATGGAGTCAATGCCAATGGTGAATGGGTAAAC
SP023 amino acid (SEQ ID NO:38)
DEQKIKQAEAEVESKQAEATRLKKIKTDREEAEEEAKRRADAKEQGKPKGRAKRGVPGELATPDKKEND AKSSDΞSVGEETLPSPSLKPEKKVAEAEKKVEEAKKKAEDQKEEDRRNYPTNTYKTLELEIAESDVEVK KAE ELVKEEAKEPRNEEKVKQAKAEVESKKAEATRLEKIKTDRKKAEEEAKRKAAEEDKVKEKPAEQP QPAPAPKAEKPAPAPKPENPAEQPKAEKPADQQAEEDYARRSEEEYNRLTQQQPPKTEKPAQPSTPKTG WKQENGM YFYNTDGSlATG LQNNGΞWYYLNSNGAMATG LQN GS YYLNANGS ATG LQNISIGSWY YLNANGSI^TGWLQYNGS YYLNANGSMATG LQYNGS YYLNA GDMATG VKDGDT YYLEASGAMK ASQWFKVSDK YYV GSGALAVNTTVDGYGVNANGEWVN
SP025 nucleotide (SEQ ID NO:39)
CTGTGGTGAGGAAGAAACTAAAAAGACTCAAGCAGCACAACAGCCAAAACAACAAACGACTGTACAACA AATTGCTGTTGGAAAAGATGCTCCAGACTTCACATTGCAATCCATGGATGGCAAAGAAGTTAAGTTATC TGATTTTAAGGGTAAAAAGGTTTACTTGAAGTTTTGGGCTTCATGGTGTGGTCCATGCAAGAAAAGTAT GCCAGAGTTGATGGAACTAGCGGCGAAACCAGATCGTGATTTCGAAATTCTTACTGTCATTGCACCAGG AATTCAAGGTGAAAAAACTGTTGAGCAATTCCCACAATGGTTCCAGGAACAAGGATATAAGGATATCCC AGTTCTTTATGATAC AAAGC.AACCACTTCCAAGCTTATCAAATTCGAAGCATTCCTACAGAATATT
SP025 amino acid (SEQ ID NO:40)
CGEEETKKTQAAQQPKQQTTVQQIAVGKDAPDFTLQSMDGKEVKLSDFKGKKVYLKF AS CGPCKKSM PELMELAAKPDRDFEILTVIAPGIQGEKTVEQFPQWFQEQGYKDIPVLYDTKATTSKLIKFEAFLQNI
SP028 nucleotide (SEQ ID NO:41)
GACTTTTAACAATAAAACTATTGAAGAGTTGCACAATCTCCTTGTCTCTAAGGAAATTTCTGCAACAGA ATTGACCCAAGCAACACTTGAAAATATCAAGTCTCGTGAGGAAGCCCTCAATTCATTTGTCACCATCGC TGAGGAGCAAGCTCTTGTTCAAGCTAAAGCCATTGATGAAGCtGGAATTGATGCTGACAATGTCCTTTC AGGAATTCCACTTGCTGTTAAGGATAACATCTCTACAGACGGTATTCTCACAACTGCTGCCTCAAAAAT GCTCTACAACTATGAGCCAATCTTTGATGCGACagCTgTTGCCAATGCAAAAACCAAGGGCATGATTGT CGTTGGAAAGACCAACATGGACGAATTTGCTATGGGTGGTTCAGGtGAAACTTCACACTACGGAGCAAC TAAAAACGCTTGGAACCACAGCAAGGTTCCTGGTGGGTCATCAAGTGGTTCTGCCGCAGCTGTAGCCTC AGGACAAGTTCGCTTGTCACTTGGTTCTGATACTGGTGGTTCCATCCGCCAACCTGCTGCCTTCAACGG AATCGTTGGTCTCAAACCAACCTACGGAACAGTTTCACGTTTCGGTCTCATTGCCTTTGGTAGCTCATT AGACCAGATTGGACCTTTTGCTCCTACTGTTAAGGAAAATGCCCTCTTGCTCAACGCTATTGCCAGCGA AGATGCTAAAGACTCTACTTCTGCTCCTGTCCGCATCGCCGACTTTACTTCAAAAATCGGCCAAGACAT CAAGGGTATGAAAATCGCTTTGCCTAAGGAATACCTAGGCGAAGGAATTGATCCAGAGGTTAAGGAAAC AATCTTAAACGCGGCCAAACACTTTGAAAAATTGGGTGCTATCGTCGAAGAAGTCAGCCTTCCTCACTC TAAATACGGTGTTGCCGTTTATTACATCATCGCTTCATCAGAAGCTTCATCAAACTTGCAACGCTTCGA CGGTATCCGTTACGGCTATCGCGCAGAAGATGCAACCAACCTTGATGAAATCTATGTAAACAGCCGAAG CCAAGGTTTTGGTGAAGAGGTAAAACGTCGTATCATGCTGGGTACTTTCAGTCTTTCATCAGGTTACTA TGATGCCTACTACAAAAAGGCTGGTCAAGTCCGTACCCTCATCATTCAAGATTTCGAAAAAGTCTTCGC GGATTACGATTTGATTTTGGGTCCAACTGCTCCAAGTGTTGCC1ΑTGACTTGGATTCTCTCAACCATGA CCCAGTTGCCATGTACTTAGCCGACCTATTGACCATACCTGTAAACTTGGCAGGACTGCCTGGAATTTC GATTCCTGCTGGATTCTCTCAAGGTCTACCTGTCGGACTCCAATTGATTGGTCCCAAGTACTCTGAGGA AACCATTTACCAAGCTGCTGCTGCTTTTGAAGCAACAACAGACTACCACAAACAACAACCCGTGATTTT TGGAGGTGACAAC
SP028 amino acid (SEQ ID NO: 42)
TFNNKTIEELHN LVSKEISATELTQATLENIKSREEALNSFVTIAEEQALVQAKAIDEAGIDADNVLS GIPLAVKDNISTDGILTTAASKMLYNYEPIFDATAVANAKTKGMIWGKTNMDEFAMGGΞGETΞHYGAT K AWNHSKVPGGSSSGSAAAVASGQVRLS GSDTGGΞIRQPAAFNGIVGLKPTYGTVSRFGLIAFGSSL Table 1 5
DQIGPFAPTVKENA L NAIASEDAKDSTSAPVRIADFTSKIGQDIKGMKIALPKEYLGEGIDPEVKET ILNAAKHFEK GAIVEEVΞLPHSKYGVAVYYIIASSEASSN QRFDGIRYGYRAEDATNLDEIYVNSRS QGFGEEλKRRIMLGTFSLSSGYYDAYYKKAGQVRTLIIQDFEKVFADYDLILGPTAPSVAYDLDSLNHD PVAMYLADLLTIPVNLAGLPGISIPAGFSQGLPVGLQLIGPKYSEETIYQAAAAFEATTDYHKQQPVIF GGDN
SP030 nucleotide (SEQ ID NO:43)
CTTTACAGGTAAACAACTACAAGTCGGCGACAAGGCGCTTGATTTTTCTCTTACTACAACAGATCTTTC TAAAAAATCTCTGGCTGATTTTGATGGCAAGAAAAAAGTCTTGAGTGTCGTTCCTTCTATCGATACAGG CATCTGCTCAACTCAAACACGTCGTTTTAATGAAGAATTGGCTGGACTGGACAACACGGTCGTATTGAC TGTTTCAATGGACCTACCTTTTGCTCAAAAACGTTGGTGCGGTGCTGAAGGCCTTGACAATGCCATTAT GCTTTCAGACTACTTTGACCATTCTTTCGGGCGCGATTATGCCCTCTTGATCAACGAATGGCACCTATT AGCACGCGCAGTCTTTGTCCTCGATACTGACAATACGATTCGCTACGTTGAATACGTGGATAATATCAA TTCTGAGCCAAACTTCGAA
SP030 amino acid (SEQ ID NO:44)
FTGKQLQVGDKALDFSLTTTDLSKKSLADFDGKKKVLSWPSIDTGICSTQTRRFNEELAGLDNTWLT VSMDLPFAQKRWCGAEG DNAIMLSDYFDHSFGRDYALLINEWHLLARAVFVLDTDNTIRYVEYVDNIN SEPNFE
SP031 nucleotide (SEQ ID NO:45)
CCAGGCTGATACAAGTATCGCAGACATTCAAAAAAGAGGCGAACTGGTTGTCGGTGTCAAACAAGACGT TCCCAATTTTGGTTACAAnGATCCCAAGACCGGTACTTATTCTGGTATCGAAaCCGACTTGGCCAAGAT GGTAGCTGATGAACTCAAGGTCAAGATTCGCTATGTGCCGGTTACAGCACAAACCCGCGGCCCCCTTCT AGACAATGAACAGGTCGATATGGATATCGCGACCTTTACCATCACGGACGAACGCAAAAAACTCTACAA CTTTACCAGTCCCTACTACACAGACGCTTCTGGATTTTTGGTCAATAAATCTGCCAAAATCAAAAAGAT TGAGGACCTAAACGGCAAAACCATCGGAGTCGCCCAAGGTTCTATCACCCAACGCCTGATTACTGAACT GGGTAAAAAGAAAGGTCTGAAGTTTAAATTCGTCGAACTTGGTTCCTACCCAGAATTGATTACTTCCCT GCACGCTCATCGTATCGATACCTTTTCCGTTGACCGCTCTATTCTATCTGGCTACACTAGTAAACGGAC AGCACTACTAGATGATAGTTTCAAGCCATCTGACTACGGTATTGTTACCAAGAAATCAAATACAGAGCT CAACGACTATCTTGATAACTTGGTTACTAAATGGAGCAAGGATGGTAGTTTGCAGAAACTTTATGACCG TTACAAGCTCAAACCATCTAGCCATACTGCAGAT
SP031 amino acid (SEQ ID NO:46)
QADTSIADIQKRGELWGVKQDVPNFGYXDPKTGTYSGIETDLAKMVADELKV IRYVPVTAQTRGPLL DNEQVDMDIATFTITDERKKLYNFTSPYYTDASGFLV KSAKIKKIEDLNGKTIGVAQGSITQRLITEL GKKKGLKFKFVELGSYPELITSLHAHRIDTFSVDRSILSGYTSKRTALLDDSFKPSDYGIVTKKSNTEL NDYLDNLVTKWSKDGSLQKLYDRYK KPSSHTAD
SP032 nucleotide (SEQ ID NO:47)
GTCTGTATCATTTGAAAACAAAGAAACAAACCGTGGTGTCTTgACTTTCACTATCTCTCAAGACCAAAT CAAACCAGAATTGGACCGTGTCTTCAAGtCAGTGAAGAAATCTCTTAATGTTCCAGGTTTCCGTAAAGG TCACCTTCCACGCCCTATCTTCGACCAAAAATTTGGTGAAGAAGCTCTTTATCAAGATGCAATGAACGC ACTTTTGCCAAACGCTTATGAAGCAGCTGTAAAAGAAGCTGGTCTTGAAGTGGTTGCCCAACCAAAAAT TGACGTAACTTCAATGGAAAAAGGTCAAGACTGGGTTATCACTGCTGAAGTCGTTACAAAACCTGAAGT AAAATTGGGTGACTACAAAAACCTTGAAGTATCAGTTGATGTAGAAAAAGAAGTAACTGACGCTGATGT CGAAGAGCGTATCGAACGCGAACGCAACAACCTGGCTGAATTGGTTATCAAGGAAGCTGCTGCTGAAAA CGGCGACACTGTTGTGATCGACTTCGTTGGTTCTATCGACGGTGTTGAATTTGACGGTGGAAAAGGTGA AAACTTCTCACTTGGACTTGGTTCAGGTCAATTCATCCCTGGTTTCGAAGACCAATTGGTAGGTCACTC AGCTGGCGAAACCGTTGATGTTATCGTAACATTCCCAGAAGACTACCAAGCAGAAGACCTTGCAGGTAA AGAAGCTAAATTCGTGACAACTATCCACGAAGTAAAAGCTAAAGAAGTTCCGGCTCTTGACGATGAACT TGCAAAAGACATTGATGAAGAAGTTGAAACACTTGCTGACTTGAAAGAAAAATACAGCAAAGAATTGGC TGCTGCTAAAGAAGAAGCTTACAAAGATGCAGTTGAAGGTGCAGCAATTGATACAGCTGTAGAAAATGC TGAAATCGTAGAACTTCCAGAAGAAATGATCCATGAAGAAGTTCACCGTTCAGTAAATGAATTCCTTGG GAATTTGCAACGTCAAGGGATCAACCCTGACATGTACTTCCAAATCACTGGAACTACTCAAGAAGACCT TCACAACCAATACCAAGCAGAAGCTGAGTCACGTACTAAGACTAACCTTGTTATCGAAGCAGTTGCCAA AGCTGAAGGATTTGATGCTTCAGAAGAAGAAATCCAAAAAGAAGTTGAGCAATTGGCAGCAGACTACAA Table 1 58
CATGGAAGTTGCACAAGTTCAAAACTTGCTTTCAGCTGACATGTTGAAACATGATATCACTATCAAAAA AGCTGTTGAATTGATCACAAGCACAGCAACAGTAAAA
SP032 amino acid (SEQ ID NO:48)
SVSFENKETNRGV TFTISQDQIKPELDRVFKSVKKS NVPGFRKGHLPRPIFDQKFGEEA YQDAMNA L PNAYEAAVKEAGLEWAQPKIDVTSMEKGQDWVITAEWTKPEVKLGDYKNLEVSVDVEKEVTDADV EERIERERNNLAE VIKEAAAENGDTWIDFVGSIDGVEFDGGKGENFSLG GΞGQFIPGFEDQLVGHS AGETVDVIVTFPEDYQAEDLAGKEAKFVTTIHEVKAKEVPALDDELAKDIDEEVETLADLKEKYSKELA AAKEEAYKDAVEGAAIDTAVENAEIVELPEEMIHEEVHRSVNEFLGNLQRQGINPDMYFQITGTTQEDL HNQYQAEAESRTKTNLVIEAVAKAEGFDASEEEIQKΞVEQLAADYNMEVAQVQNLLSADMLKHDITIKK AVELITSTA VK
SP033 nucleotide (SEQ ID NO:49)
TGGTCAAAAGGAAAGTCAGACAGGAAAGGGGATGAAAATTGTGACCAGTTTTTATCCTATCTACGCTAT GGTTAAGGAAGTATCTGGTGACTTGAATGATGTTCGGATGATTCAGTCAAGTAGTGGTATTCACTCCTT TGAACCTTCGGCAAATGATATCGCAGCCATCTATGATGCAGATGTCTTTGTTTACCATTCTCATACACT CGAATCTTGGGCAGGAAGTCTGGATCCAAATCTAAAAAAATCCAAAGTGAAGGTCTTAGAGGCTTCTGA GGGAATGACCTTGGAACGTGTCCCTGGACTAGAGGATGTGGAAGCAGGGGATGGAGTTGATGAAAAAAC GCTCTATGACCCTCACACATGGCTAGATCCTGAAAAAGCTGGAGAAGAAGCCCAAATTATCGCTGATAA ACTTTCAGAGGTGGATAGTGAGCATAAAGAGACTTATCAAAAAAATGCGCAACCTTTATCAAAAAAGCT CAGGAAT
SP033 amino acid (SEQ ID NO:50)
GQKESQTGKGMKIVTSFYPIYA VKEVSGDLNDVRMIQSSSGIHSFEPSANDIAAIYDADVFΛT/HSHTL ESWAGSLDPNLKKSKVKVLEASEGMTLΞRVPGLΞDVΞAGDGVDEKTLYDPHT LDPEKAGEEAQIIADK LSEVDSEHKETYQKNAQPLSKKLRN
SP034 nucleotide (SEQ ID NO: 51)
GAAGGATAGATATATTTTAGCATTTGAGACATCCTGTGATGAGACCAGTGTCGCCGTCTTGAAAAACGA CGATGAGCTCTTGTCCAATGTCATTGCTAGTCAAATTGAGAGTCACAAACGTTTTGGTGGCGTAGTGCC CGAAGTAGCCAGTCGTCACCATGTCGAGGTCATTACAGCCTGTATCGAGGAGGCATTGGCAGAAGCAGG GATTACCGAAGAGGACGTGACAGCTGTTGCGGTTACCTACGGACCAGGCTTGGTCGGAGCCTTGCTAGT TGGTTTGTCAGCTGCCAAGGCCTTTGCTTGGGCTCACGGACTTCCACTGATTCCTGTTAATCACATGGC TGGGCACCTCATGGCAGCTCAGAGTGTGGAGCCTTTGGAGTTTCCCTTGCTAGCCCTCTTGGTCAGCGG CGGACACACAGAGTTGGTTTATGTTTCGGAGGCAGGAGATTATAAGATTGTTGGGGAAACCCGTGATGA TGCGGTTGGTGAGGCTTATGATAAGGTCGGCCGTGTCATGGGCTTGACCTATCCTGCAGGTCGTGAGAT TGACGAGCTGGCTCATCAGGGGCAGGATATTTATGATTTCCCCCGTGCCATGATTAAGGAAGATAATCT GGAGTTCTCCTTCTCAGGTTTGAAATCTGCCTTTATCAATCTTCATCACAATGCCGAGCAAAAGGGAGA AAGCCTGTCTACAGAAGATTTGTGTGCTTCCTTCCAAGCAGCAGTTATGGACATTCTCATGGCAAAAAC CAAGAAGGCTTTGGAGAAATATCCTGTTAAAATCCTAGTTGTGGCAGGTGGTGTGGCAGCCAATAAAGG TCTCAGAGAACGCCTAGCAGCCGAAATCACAGATGTCAAGGTTATCATCCCCCCTCTGCGACTCTGCGG AGACAATGCAGGTATGATTGCCTATGCCAGCGTCAGCNAGTGGAACAAAGAAAACTTCGCAGGCTGGGA CCTCAATGCCAAACCAAGTCTTGCCTTTGATACCATGGAA
SP034 amino acid (SEQ ID NO:52)
KDRYILAFETSCDETSVAVLKNDDELLSNVIASQIESHKRFGGWPEVASRHHVEVITACIEEALAEAG ITEEDVTAVAVTYGPG VGALLVGLSAAKAFA AHGLPLIPVNHMAGHLMAAQSVEPLEFPLLALLVSG GHTELVYVSEAGDYKIVGETRDDAVGEAYDKVGRVMG TYPAGREIDELAHQGQDIYDFPRAMIKEDNL EFSFΞGLKSAFIN HHNAEQKGESLSTEDLCASFQAAV DI MAKTKKALEKYPVKILWAGGVAANKG LRERLAAEITDV VIIPPLRLCGDNAGMIAYASVSXWNKENFAG DLNAKPSLAFDT E
SP035 nucleotide (SEQ ID NO:53)
GGTAGTTAAAGTTGGTATTAACGGTTTCGGACGTATCGGTCGTCTTGCTTTCCGTCGTATCCAAAACGT AGAAGGTGTTGAAGTTACACGCATCAACGACCTTACAGATCCAGTTATGCTTGCACACTTGTTGAAATA CGACACAACTCAAGGTCGTTTCGACGGTACTGTTGAAGTTAAAGAAGGTGGATTTGAAGTTAACGGTAA ATTCATCAAAGTTTCTGCTGAACGTGATCCAGAACAAATCGACTGGGCTACTGACGGTGTAGAAATCGT TCTTGAAGCTACTGGTTTCTTTGCTAAGAAAGAAGCAGCTGAAAAACACCTTAAAGGTGGAGCTAAAAA Table 1 59
AGTTGTTATCACTGCTCCTGGTGGAAACGACGTTAAAACAGTTGTATTCAACACTAACCACGACGTTCT TGACGGTACTGAAACAGTTATCTCAGGTGCTTCATGTACTACAAACTGCTTGGCTCCAATGGCTAAAGC TCTTCAAGACAACTTTGGTGTTGTTGAAGGATTGATGACTACTATCCACGCTTACACTGGTGACCAAAT GATCCTTGACGGACCACACCGTGGTGGTGACCTTCGCCGTGCTCGCGCTGGTGCTGCAAACATCGTTCC TAACTCAACTGGTGCTGCAAAAGCTATCGGTCTTGTAATCCCAGAATTGAATGGTAAACTTGACGGATC TGCACAACGCGTTCCAACTCCAACTGGATCAGTTACTGAATTGGTAGCAGTTCTTGAAAAGAACGTTAC TGTTGATGAAGTGAACGCAGCTATGAAAGCAGCTTCAAACGAATCATACGGTTACACAGAAGATCCAAT CGTATCTTCAGATATCGTAGGTATGTCTTACGGTTCATTGTTTGACGCAACTCAAACTAAAGTTCTTGA CGTTGACGGTAAACAATTGGTTAAAGTTGTATCATGGTACGACAACGAAATGTCATACACTGCACAACT TGTTCGTACTCTTGGAATACTTCGCAAAAATTGC
SP035 amino acid (SEQ ID NO:54)
VV VGINGFGRIGRIAFRRIQ1WEGVEVTRINDLTDPVMLAHLLKYDTTQGRFDGTVEVKEGGFEV GK FIKVSAERDPEQID ATDGVEIV EATGFFAKKEAAEKHLKGGAKKVVITAPGGNDVKTVVFNTNHDVL DGTETVISGASCTTNCLAPMAKALQDNFGWEGL TTIHAYTGDQMILDGPHRGGDLRRARAGAANIVP NSTGAAKAIGLVIPELNGKLDGSAQRVPTPTGSVTELVAVLEK VTVDEVNAAMKAASNESYGYTEDPI VSSDIVGMSYGSLFDATQTKVLDVOGKQLVKWS YDNEMSYTAQLVRTLGILRKNC
SP036 nucleotide (SEQ ID NO:55)
TTCTTACGAGTTGGGACTGTATCAAGCTAGAACGGTTAAGGAAAATAATCGTGTTTCCTATATAGATGG AAAACAAGCGACGCAAAAAACGGAGAATTTGACTCCTGATGAGGTTAGCAAGCGTGAAGGAATCAATGC TGAGCAAATCGTCATCAAGATAACAGACCAAGGCTATGTCACTTCACATGGCGACCACTATCATTATTA CAATGGTAAGGTTCCTTATGACGCTATCATCAGTGAAGAATTACTCATGAAAGATCCAAACTATAAGCT AAAAGATGAGGATATTGTTAATGAGGTCAAGGGTGGATATGTTATCAAGGTAGATGGAAAATACTATGT TTACCTTAAGGATGC^GCCCACGCGGATAACGTCCGTACAAAAGAGGAAATCAATCGACAAAAACAAGA GCATAGTCAACATCGTGAAGGTGGAACTCCAAGAAACGATGGTGCTGTTGCCTTGGCACGTTCGCAAGG ACGCTATACTACAGATGATGGTTATATCTTTAATGCTTCTGATATCATAGAGGATACTGGTGATGCTTA TATCGTTCCTCATGGAGATCATTACCATTACATTCCTAAGAATGAGTTATCAGCTAGCGAGTTGGCTGC TGCAGAAGCCTTCCTATCTGGTCGAGGAAATCTGTCAAATTCAAGAACCTATCGCCGACAAAATAGCGA TAACACTTCAAGAACAAACTGGGTACCTTCTGTAAGCAATCCAGGAACTACAAATACTAACACAAGCAA CAACAGCAACACTAACAGTCAAGCAAGTCAAAGTAATGACATTGATAGTCTCTTGAAACAGCTCTACAA ACTGCCTTTGAGTCAACGACATGTAGAATCTGATGGCCTTGTCTTTGATCCAGCACAAATCACAAGTCG AACAGCTAGAGGTGTTGCAGTGCCACACGGAGATCATTACCACTTCATCCCTTACTCTCAAATGTCTGA ATTGGAAGAACGAATCGCTCGTATTATTCCCCTTCGTTATCGTTCAAACCATTGGGTACCAGATTCAAG GCCAGAACAACCAAGTCCACAACCGACTCCGGAACCTAGTCCAGGCCCGCAACCTGCACCAAATCTTAA AATAGACTCAAATTCTTCTTTGGTTAGTCAGCTGGTACGAAAAGTTGGGGAAGGATATGTATTCGAAGA AAAGGGCATCTCTCGTTATGTCTTTGCGAAAGATTTACCATCTGAAACTGTTAAAAATCTTGAAAGCAA GTTATCAAAACAAGAGAGTGTTTCACACACTTTAACTGCTAAAAAAGAAAATGTTGCTCCTCGTGACCA AGAATTTTATGATAAAGCATATAATCTGTTAACTGAGGCTCATAAAGCCTTGTTTGNAAATAAGGGTCG TAATTCTGATTTCCAAGCCTTAGACAAATTATTAGAACGCTTGAATGATGAATCGACTAATAAAGAAAA ATTGGTAGATGATTTATTGGCATTCCTAGCACCAATTACCCATCCAGAGCGACTTGGCAAACCAAATTC TCAAATTGAGTATACTGAAGACGAAGTTCGTATTGCTCAATTAGCTGATAAGTATACAACGTCAGATGG TTACATTTTTGATGAACATGATATAATCAGTGATGAAGGAGATGCATATGTAACGCCTCATATGGGCCA TAGTCACTGGATTGGAAAAGATAGCCTTTCTGATAAGGAAAAAGTTGCAGCTCAAGCCTATACTAAAGA AAAAGGTATCCTACCTCCATCTCCAGACGCAGATGTTAAAGCAAATCCAACTGGAGATAGTGCAGCAGC TATTTACAATCGTGTGAAAGGGGAAAAACGAATTCCACTCGTTCGACTTCCATATATGGTTGAGCATAC AGTTGAGGTTAAAAACGGTAATTTGATTATTCCTCATAAGGATCATTACCATAATATTAAATTTGCTTG GTTTGATGATCACACATACAAAGCTCCAAATGGCTATACCTTGGAAGATTTGTTTGCGACGATTAAGTA CTACGTAGAACACCCTGACGAACGTCCACATTCTAATGATGGATGGGGCAATGCCAGTGAGCATGTGTT AGGCAAGAAAGACCACAGTGAAGATCCAAATAAGAACTTCAAAGCGGATGAAGAGCCAGTAGAGGAAAC ACCTGCTGAGCCAGAAGTCCCTCAAGTAGAGACTGAAAAAGTAGAAGCCCAACTCAAAGAAGCAGAAGT TTTGCTTGCGAAAGTAACGGATTCTAGTCTGAAAGCCAATGCAACAGAAACTCTAGCTGGTTTACGAAA TAATTTGACTCTTCAAATTATGGATAACAATAGTATCATGGCAGAAGCAGAAAAATTACTTGCGTTGTT AAAAGGAAGTAATCCTTCATCTGTAAGTAAGGAAAAAATAAAC
SP036 amino acid (SEQ ID NO:56)
SYELG YQARTVKENNRVSYIDGKQATQKTENLTPDEVSKREGINAEQIVIKITDQGYVTSHGDHYHYY NGKVPYDAIIΞEΞ KDPNYKLKΏEDIVNF^KGGYVIKVDGKYYVYLKDAAHADNVRTKEEINRQKQE Table 1 60
HSQHREGGTPRNDGAVALARSQGRYTTDDGYIFNASDIIEDTGDAYIVPHGDHYHYIPKNELSASELAA AEAFLSGRGNLSNSRTYRRQNSDNTSRTNWVPSVSNPGTTNTNTSN SNTNSQASQSNDIDΞLLKQLYK PLSQRHVESDGLVFDPAQITSRTARGVAVPHGDHYHFIPYSQMSELEERIARIIPLRYRSNH VPDSR PEQPSPQPTPEPSPGPQPAPNLKIDSNSSLVSQLVRKVGEGYVFEEKGISRYVFAKDLPSETVKNLESK SKQESVSHTLTAKKENVAPRDQEFYDKAYNL TEAHKALFXNKGRNSDFQA DKLLERLNDESTNKEK LVDDLLAFLAPITHPERLGKPNSQIEYTEDEVRIAQLADKYTTSDGYIFDEHDIISDEGDAYVTPHMGH SH IGKDSLSDKEKVAAQAYTKEKGI PPSPDADVKANPTGDSAAAIYNRVKGEKRIPLVR PYMVEKT VEVKNGNLIIPHKDHYHNIKFAWFDDHTYKAPNGYTLEDLFATIKYYVEHPDERPHSNDG GNASEHVL GKKDHSEDPNKNFKADEEPVEETPAEPF/PQVETEKVEAQLKEAEV LAKVTDSSLKANATETLAGLRN NLTLQIMDNNSIMAEAEKL A LKGSNPSSVSKEKIN
SP038 nucleotide (SEQ ID NO:57)
TACTGAGATGCATCATAATCTAGGAGCTGAAAAGCGTTCAGCAGTGGCTACTACTATCGATAGTTTTAA GGAGCGAAGTCAAAAAGTCAGAGCACTATCTGATCCAAATGTGCGTTTTGTTCCCTTCTTTGGCTCTAG TGAATGGCTTCGTTTTGACGGTGCTCATTCTGCGGTATTAGCTGAGAAATACAATCGTTCCTACCGTCC TATCTTTTAGGACAGGGGGGAGCTGCATCGCTTAACCAATATTTTGGAATGCAACAGATGTTACCACA GCTGGAGAATAAACAAGTTGTGTATGTTATCTCACCTCAGTGGTTCAGTAAAAATGGCTATGATCCAGC AGCCTTCCAGCAGTATTTTAATGGAGACCAGTTGACTAGTTTTCTGAAACATCAATCTGGGGATCAGGC TAGTCAATATGCAGCGACTCGCTTACTGCAACAGTTCCCAAACGTAGCTATGAAGGACCTGGTTCAGAA GTTGGCAAGTAAAGAAGAATTGTCGACAGCAGACAATGAAATGATTGAATTATTGGCTCGTTTTAATGA ACGCCAAGCTTCCTTTTTTGGTCAGTTTTCGGTTAGAGGCTATGTTAACTACGATAAGCATGTAGCTAA GTATTTAAAAATCTTGCCAGACCAGTTTTCTTATCAGGCAATAGAAGATGTTGTCAAAGCAGATGCTGA AAAAAATACTTCCAATAATGAGATGGGAATGGAAAATTATTTCTATAATGAGCAGATCAAGAAGGATTT GAAGAAATTAAAGGATTCTCAGAAAAGCTTTACCTATCTCAAGTCGCCAGAGTATAATGNNTTGCAGTT GGTTTTAACACAGTTTTCTAAATCTAAGGTAAACCCGATTTTTATCATTCCACCTGTTAATAAAAAATG GATGNACTATGCTGGTCTACGAGAGGATATGTACCAACAAACGGTGCAGAAGATTCGCTACCAGTTAGA AAGTCAAGGTTTTACCAATATAGCAGATTTTTCTAAGGACGGCGGGGAGCCTTTCTTTATGAAGGACAC CATTCACCTTGGTTGGTTGGGTTGGTTGGCTTTTGACAAGGCAGTTGATCCTTTCCTATCCAATCCCAC ACCAGCTCCGACTTACCATCTGAATGAGCGCTTTTTCAGCAAAGATTGGGCGACTTATGATGGAGATGT
CAAAGAA
SP038 amino acid (SEQ ID NO:58) EMHHNLGAEKRSAVATTIDSFKERΞQKVRALSDPNVRFVPFFGSSEWLRFDGAHSAVLAEKYNRSYRP YLLGQGGAASLNQYFGMQQMLPQLENKQWΎVISPQ FSKNGYDPAAFQQYFNGDQLTSFLKHQSGDQA SQYAATRLLQQFP VAMKDLVQKLASKEELSTADNEMIΞ LARFNERQASFFGQFSVRGYVNYDKHVAK YLKILPDQFSYQAIEDWKADAEKNTSNNE GMEMΎFY EQIKKDLKKLKDSQKSFTYLKSPEYNXLQL VLTQFSKΞKV PIFIIPPVNKKWMXYAGLREDMYQQTVQKIRYQLESQGFTNIADFSKDGGEPFFMKDT IHLGWLGWLAFDKAVDPFLSNPTPAPTYHLNERFFSKDWATYDGDVKE
SP039 nucleotide (SEQ ID NO:59)
GGTTTTGAGAAAGTATTTGCAGGGGGCCCTGATTGAGTCGATTGAGCAAGTGGAAAATGACCGTATTGT GGAAATTACAGTTTCCAATAAAAACGAGATTGGAGACCATATCCAGGCTACCTTGATTATCGAAATTAT GGGGAAACACAGTAATATTCTACTGGTCGATAAAAGCAGTCATAAAATCCTCGAAGTTATCAAACACGT CGGCTTTTCACAAAATAGCTACCGCACCTTACTTCCAGGATCGACCTATATCGCTCCGCCAAGTACAAA ATCTCTCAATCCTTTTACTATCAAGGATGAAAAGCTCTTTGAAATCCTGCAAACCCAAGAACTAACAGC AAAAATCTTCAAAGCCTCTTTCAAGGTCTGGGACGCGATACGGCAAATGAATTGGAAAGGATACTGGT TAGTGAAAAACTTTCCGCTTTCCGAAATTTTTTCAATCAAGAAACCAAGCCATGCTTGACTGAGACTTC CTTCAGTCCAGTTCCTTTTGCAAATCAGGTGGGAGAGCCTTTTGCAAATCTTTCTGATTTGTTGGACAC CTACTATAAGGATAAGGCTGAGCGCGACCGCGTCAAACAGCAGGCCAGTGAACTGATTCGTCGTGTTGA AATGAACTTCAGAAAAACCGACACAAACTCAAAAAACAGGAAAAAGAGTTACTGGCGACAGACAACGC GAAGAATTTCGTCAAAAAGGAGAATTGCTGACAACCTTCCTCCACCAAGTGCCTAACGACCAAGACCA GGTTATCCTAGACAACTACTATACCAACCAACCTATCATGATTGCGCTTGATAAGGCTCTGACTCCCAA CCAGAATGCCCAACGCTATTTTAAACGGTATCAGAAACTCAAAGAAGCTGTCAAATACTTGACTGATTT GATTGAAGAAACCAAAGCCACTATTCTCTATCTGGAAAGTGTAGAAACCGTCCTCAACCAAGCTGGACT GGAAGAAATCGCTGAAATCCGTGAAGAATTGATTCAAACAGGTTTTATCCGCAGAAGACAACGGGAGAA AATCCAGAAACGCAAAAAACTAGAACAATATCTAGCAAGCGATGGCAAAACCATCATCTATGTCGGACG AAACAATCTTCAAAATGAGGAATTGACCTTTAAAATGGCCCGCAAGGAGGAACTTTGGTTCCATGCTAA GGACATTCCTGGAAGCCATGTTGTCATCTCAGGAAATCTTGACCCATCTGATGCAGTCAAGACAGACGC Table 1 61
AGCAGAGTTAGCTGCCTACTTCTCTCAAGGGCGCCTGTCGAATCTGGTGCAGGTAGATATGATTGAAGT CAAAAAACTCAATAAACCAACTGGTGGAAAACCCGGCTTTGTCACTTACACAGGACAAAAGACCCTCCG CGTCACACCAGACTCCAAAAAAATTGCATCCATGAAAAAATCC
SP039 amino acid (SEQ ID NO:60)
VLRKYLQGALIESIEQVENDRIVEITVSNKNEIGDHIQATLIIEIMGKHSNI LVDKSSHKI EVIKHV GFSQNSYRTLLPGSTYIAPPSTKSLNPFTIKDEK FEILQTQELTAK LQSLFQGLGRDTANELERILV SEKLSAFRNFFNQETKPCLTETSFSPVPFANQVGEPFANLSD LDTYYKDKAERDRVKQQASELIRRVE NELQKNRHK KKQEKELLATDNAEEFRQKGELLTTFLHQVPNDQDQVILDNYYTNQPIMIALDKALTPN QNAQRYFKRYQKLKEAVKYLTDLIEETKATILYLESVETVLNQAGLEEIAEIREELIQTGFIRRRQREK IQKRKKLEQYLAΞDGKTIIYVGRNNLQNEELTFKMARKEEL FHAKDIPGSHWISGNLDPSDAVKTDA AELAAYFSQGRLSNLVQVDMIEVKKLNKPTGGKPGFVTYTGQKTLRVTPDSKKIASMKKS
SP040 nucleotide (SEQ ID NO: 61)
GACAACATTTACTATCCATACAGTAGAGTCAGCACCAGCAGAAGTGAAAGAAATTCTTGAAACAGTAGA AAAAGACAACAATGGCTATATTCCCAACCTAATCGGTCTCTTGGCCAATGCCCCGACTGTTTTAGAAGC
CTACCAAATTGTCTCATCTATCCACCGTCGCAACAGCCTGACACCCGTTGAGCGTGAAGTGGTGCAAAT CACGGCAGCCGTGACCAATGGTTGTGCCTTCTGTGTCGCAGGTCACACAGCCTTTTCCATCAAACAAAT CCAGATGAATGATGACTTGATTCAAGCTCTTCGCAATCGTACTCCAATTGAAACAGATCCTAAATTGGA ACCCTAGCTAAGTTTACCTTGGCAGTTATCAATACCAAGGGTCGTGTAGGAGATGAAGCCTTGTCTGA GTTTTTAGAAGCTGGCTACACTCAACAAAATGCCTTGGATGTGGTTTTTGGTGTCAGCCTAGCAATCCT CTGTAACTATGCCAACAACTTAGCTAATACACCAATTAATCCAGAATTGCAACCTTATGCC
SP040 amino acid (SEQ ID NO:62)
TTFTIHTVESAPAΞVKEILETVEKDNNGYIPNLIGLLANAPTVLEAYQIVΞSIHRRNSLTPVEREVVQI TAAVTNGCAFCVAGHTAFSIKQIQMNDDLIQALRNRTPIETDPKLDTLAKFTLAVINTKGRVGDEALSE FLEAGYTQQNALDWFGVSLAILCNYANNLANTPINPELQPYA
SP041 nucleotide (SEQ ID NO: 63)
GGCTAAGGAAAGAGTGGATGTACTAGCTTATAAACAGGGGTTGTTTGAAACGAGAGAGCAGGCCAAGCG AGGTGTGATGGCTGGCCTAGTCGTAGCAGTCCTTAATGGAGAACGGTTTGACAAGCCAGGAGAGAAAAT TCCAGATGACACCGAATTAAAACTCAAGGGGGAGAAACTCAAGTATGTCAGCCGTGGTGGTTTGAAACT GGAAAAGGCCTTGCAGGTCTTTGATTTGTCGGTGGATGGCGCGACTACGATTGATATCGGGGCCTCTAC TGGAGGTTTTACCGATGTCATGCTACAGAATAGTGCCAAGTTGGTCTTTGCAGTCGATGTTGGTACCAA TCAGTTGGCTTGGAAATTACGCCAAGACCCACGAGTTGTCAGCATGGAGCAGTTCAATTTCCGCTATGC TGAAAAGACTGATTTCGAGCAGGAGCCGAGCTTTGCCAGTATTGATGTGAGTTTCATTTCCCTTAGTCT GATTTTGCCAGCCTTGCACCGTGTCTTGGCTGATCAAGGTCAGGTGGTAGCACTTGTCAAACCTCAGTT TGAGGCAGGACGTGAGCAGATTGGGAAAAATGGAATTATTCGAGATGCTAAGGTTCATCAGAATGTCCT TGAATCTGTAACAGCTATGGCAGTAGAGGTAGGTTTTTCAGTCCTTGGCTTGGACTTTTCTCCCATCCA AGGTGGACATGGAAATATTGAATTTTTAGCGTATTTGAAAAAAGAAAAGTCAGCAAGCAATCAGATTCT TGCTGAGATTAAAGAAGCAGTAGAGAGGGCGCATAGTCAATTTAAAAATGAA
SP041 amino acid (SEQ ID NO:64)
AKERVDVLAYKQGLFETREQAKRGVMAGLWAVLNGERFDKPGEKIPDDTELKLKGEKLKYVSRGGLKL EKALQVFDLΞ'VDGATTIDIGASTGGFTDVMLQNSAKLVFAVDVGTNQLA KLRQDPRWS EQFNFRYA EKTDFEQEPSFASIDVSFISLSLILPALHRVLADQGQWALVKPQFEAGREQIGKNGIIRDAKVHQNVL ESVTAMAVEVGFSV GLDFΞPIQGGHGNIEF AYLKKEKSASNQI AEIKEAVERAHSQFKNE
SP042 nucleotide (SEQ ID NO: 65)
TTGTTCCTATGAACTTGGTCGTCACCAAGCTGGTCAGGTTAAGAAAGAGTCTAATCGAGTTTCTTATAT AGATGGTGATCAGGCTGGTCAAAAGGCAGAAAACTTGACACCAGATGAAGTCAGTAAGAGGGAGGGGAT CAACGCCGAACAAATNGTNATCAAGATTACGGATCAAGGTTATGTGACCTCTCATGGAGACCATTATCA TTACTATAATGGCAAGGTTCCTTATGATGCCATCATCAGTGAAGAGCTCCTCATGAAAGATCCGAATTA TCAGTTGAAGGATTCAGACATTGTCAATGAAATCAAGGGTGGTTATGTCATTAAGGTAAACGGTAAATA CTATGTNTACCTTAAGGATGCAGCTCATGCGGATAATATTCGGACAAAAGAAGAGATTAAACGTCAGAA GCAGGAACGCAGTCATAATCATAACTCAAGAGCAGATAATGCTGTTGCTGCAGCCAGAGCCCAAGGACG TTATACAACGGATGATGGGTATATCTTCAATGCATCTGATATCATTGAGGACACGGGTGATGCTTATAT CGTTCCTCACGGCGACCATTACCATTACATTCCTAAGAATGAGTTATCAGCTAGCGAGTTAGCTGCTGC Table 1 62
AGAAGCCTATTGGAATGGGAAGCAGGGATCTCGTCCTTCTTCAAGTTCTAGTTATAATGCAAATCCAGC TCAACCAAGATTGTCAGAGAACCACAATCTGACTGTCACTCCAACTTATCATCAAAATCAAGGGGAAAA CATTTCAAGCCTTTTACGTGAATTGTATGCTAAACCCTTATCAGAACGCCATGTGGAATCTGATGGCCT TATTTTCGACCCAGCGCAAATCACAAGTCGAACCGCCAGAGGTGTAGCTGTCCCTCATGGTAACCATTA CCACTTTATCCCTTATGAACAAATGTCTGAATTGGAAAAACGAATTGCTCGTATTATTCCCCTTCGTTA TCGTTCAAACCATTGGGTACCAGATTCAAGACCAGAACAACCAAGTCCACAATCGACTCCGGAACCTAG TCCAAGTCCGCAACCTGCACCAAATCCTCAACCAGCTCCAAGCAATCCAATTGATGAGAAATTGGTCAA AGAAGCTGTTCGAAAAGTAGGCGATGGTTATGTCTTTGAGGAGAATGGAGTTTCTCGTTATATCCCAGC CAAGGATCTTTCAGCAGAAACAGCAGCAGGCATTGATAGCAAACTGGCCAAGCAGGAAAGTTTATCTCA TAAGCTAGGAGCTAAGAAAACTGACCTCCCATCTAGTGATCGAGAATTTTACAATAAGGCTTATGACTT ACTAGCAAGAATTCACCAAGATTTACTTGATAATAAAGGTCGACAAGTTGATTTTGAGGCTTTGGATAA CCTGTTGGAACGACTCAAGGATGTCNCAAGTGATAAAGTCAAGTTAGTGGANGATATTCTTGCCTTCTT AGCTCCGATTCGTCATCCAGAACGTTTAGGAAAACCAAATGCGCAAATTACCTACACTGATGATGAGAT TCAAGTAGCCAAGTTGGCAGGCAAGTACACAACAGAAGACGGTTATATCTTTGATCCTCGTGATATAAC CAGTGATGAGGGGGATGCCTATGTAACTCCACATATGACCCATAGCCACTGGATTAAAAAAGATAGTTT GTCTGAAGCTGAGAGAGCGGCAGCCCAGGCTTATGCTAAAGAGAAAGGTTTGACCCCTCCTTCGACAGA CCATCAGGATTCAGGAAATACTGAGGCAAAAGGAGCAGAAGCTATCTACAACCGCGTGAAAGCAGCTAA GAAGGTGCCACTTGATCGTATGCCTTACAATCTTCAATATACTGTAGAAGTCAAAAACGGTAGTTTAAT CATACCTCATTATGACCATTACCATAACATCAAATTTGAGTGGTTTGACGAAGGCCTTTATGAGGCACC TAAGGGGTATACTCTTGAGGATCTTTTGGCGACTGTCAAGTACTATGTCGAACATCCAAACGAACGTCC GCATTCAGATAATGGTTTTGGTAACGCTAGCGACCATGTTCAAAGAAACAAAAATGGTCAAGCTGATAC CAATCAAACGGAAAAACCAAGCGAGGAGAAACCTCAGACAGAAAAACCTGAGGAAGAAACCCCTCGAGA AGAGAAACCGCAAAGCGAGAAACCAGAGTCTCCAAAACCAACAGAGGAACCAGAAGAATCACCAGAGGA ATCAGAAGAACCTCAGGTCGAGACTGAAAAGGTTGAAGAAAAACTGAGAGAGGCTGAAGATTTACTTGG AAAAATCCAGGAT /
SP042 amino acid (SEQ ID NO:66)
CSYELGRHQAGQVKKESNRVSYIDGDQAGQKAENLTPDEVSKREGINAEQXVIKITDQGY^/TSHGDHYH YYNGKVPYDAIISEELLMKDPNYQLKDSDIVNEIKGGYVIKVNGKYYVYLKDAAHADNIRTKEEIKRQK QERSHNHNSRADNAVAAARAQGRYTTDDGYIFNASDIIΞDTGDAYIVPHGDHYHYIPKNEL5ASELAAA EAY NGKQGSRPSSΞSΞYNANPAQPRLSENHNLTV PTYHQNQGENISSLLRELYAKPLSERHVESDGL IFDPAQITSRTARGVAVPHGNHYHFIPYEQMΞELEKRIARIIPLRYRSNH VPDSRPEQPSPQSTPEPS PSPQPAPNPQPAPSNPIDEK VKEAVRKVGDGYVFEENGVSRYIPAKDLSAETAAGIDSKLAKQESLSH KLGAKKTDLPSSDREFYNKAYDLLARIHQDLLDNKGRQVDFEALDNLLERLKDVXSDKVKLVXDI AFL APIRHPERLGKPNAQITYTDDEIQVAKLAGKYTTΞDGYIFDPRDITSDEGDAYVTPHMTHSH IKKDΞL SEAERAAAQAYAKEKGLTPPSTDHQDSGNTEAKGAEAIY RVKAAKKVP DR PYNLQY'T/ΞVKNGSLI IPHYDHYHNIKFE FDEGLYEAPKGYTLEDLLATVKYYVEHPNERPHSDNGFGNASDHVQRNKNGQADT NQTEKPSΞEKPQTEKPEEETPREEKPQSEKPΞSPKPTEEPEESPEESEEPQVΞTE VEEK REAEDLLG KIQD
SP043 nucleotide (SEQ ID NO: 67)
TTATAAGGGTGAATTAGAAAAAGGATACCAATTTGATGGTTGGGAAATTTCTGGTTTCGAAGGTAAAAA AGACGCTGGCTATGTTATTAATCTATCAAAAGATACCTTTATAAAACCTGTATTCAAGAAAATAGAGGA GAAAAAGGAGGAAGAAAATAAACCTACTTTTGATGTATCGAAAAAGAAAGATAACCCACAAGTAAACCA TAGTCAATTAAATGAAAGTCACAGAAAAGAGGATTTACAAAGAGAAGAGCATTCACAAAAATCTGATTC AACTAAGGATGTTACAGCTACAGTTCTTGATAAAAACAATATCAGTAGTAAATCAACTACTAACAATCC TAATAAG
SP043 amino acid (SEQ ID NO:68)
YKGELEKGYQFDG EISGFEGKKDAGYVINLSKDTFIKPVFKKIEEKKEEENKPTFDVSKKKDNPQVNH SQLNESHRKEDLQREEHSQKSDSTKDVTATVLDK NISSKSTTNNPNK
SP044 nucleotide (SEQ ID NO: 69)
GAATGTTCAGGCTCAAGAAAGTTCAGGAAATAAAATCCACTTTATCAATGTTCAAGAAGGTGGCAGTGA TGCGATTATTCTTGAAAGCAATGGACATTTTGCCATGGTGGATACAGGAGAAGATTATGATTTCCCAGA TGGAAGTGATTCTCGCTATCCATGGAGAGAAGGAATTGAAACGTCTTATAAGCATGTTCTAACAGACCG TGTCTTTCGTCGTTTGAAGGAATTGGGTGTCCAAAAACTTGATTTTATTTTGGTGACCCATACCCACAG TGATCATATTGGAAATGTTGATGAATTACTGTCTACCTATCCAGTTGACCGAGTCTATCTTAAGAAATA Table 1 63
TAGTGATAGTCGTATTACTAATTCTGAACGTCTATGGGATAATCTGTATGGCTATGATAAGGTTTTACA GACTGCTGCAGAAAAAGGTGTTTCAGTTATTCAAAATATCACACAAGGGGATGCTCATTTTCAGTTTGG GGACATGGATATTCAGCTCTATAATTATGAAAATGAAACTGATTCATCGGGTGAATTAAAGAAAATTTG GGATGACAATTCCAATTCCTTGATTAGCGTGGTGAAAGTCAATGGCAAGAAAATTTACCTTGGGGGCGA TTTAGATAATGTTCATGGAGCAGAAGACAAGTATGGTCCTCTCATTGGAAAAGTTGATTTGATGAAGTT TAATCATCACCATGATACCAACAAATCAAATACCAAGGATTTCATTAAAAATTTGAGTCCGAGTTTGAT TGTTCAAACTTCGGATAGTCTACCTTGGAAAAATGGTGTTGATAGTGAGTATGTTAATTGGCTCAAAGA ACGAGGAATTGAGAGAATCAACGCAGCCAGCAAAGACTATGATGCAACAGTTTTTGATATTCGAAAAGA CGGTTTTGTCAATATTTCAACATCCTACAAGCCGATTCCAAGTTTTCAAGCTGGTTGGCATAAGAGTGC ATATGGGAACTGGTGGTATCAAGCGCCTGATTCTACAGGAGAGTATGCTGTCGGTTGGAATGAAATCGA AGGTGAATGGTATTACTTTAACCAAACGGGTATCTTGTTACAGAATCAATGGAAAAAATGGAACAATCA TTGGTTCTATTTGACAGACTCTGGTGCTTCTGCTAAAAATTGGAAGAAAATCGCTGGAATCTGGTATTA TTTTAACAAAGAAAACCAGATGGAAATTGGTTGGATTCAAGATAAAGAGCAGTGGTATTATTTGGATGT TGATGGTTCTATGAAGACAGGATGGCTTCAATATATGGGGCAATGGTATTACTTTGCTCCATCAGGGGA A
SP044 amino acid (SEQ ID NO:70)
NV AQESSGNKIKFINVQEGGSDAIILESNGHFAMVDTGEDYDFPDGSDSRYPWREGIETSYKHVLTDR VFRRLKELGVQKLDFILVTHTHSDHIGNVDELLSTYPVDRVYLKKYSDSRITNSERLWDNLYGYDKVLQ TAAEKGVSVIQNITQGDAHFQFGDMDIQLYNYENETDSSGΞLKKI DDNΞNSLISWKVNGKKIYLGGD LDNVHGAΞDKYGPLIGKVDLMKFNHHHDTNKSNTKDFIKNLSPSLIVQTΞDSLPWKNGVDSEYVNWLXE RGIERINAASKDYDATVFDIRKDGFVNISTSYKPIPSFQAGWHKΞAYGNVWYQAPDSTGEYAVG NEIΞ GE YYFNQTGILLQNQWKK NNHWFYLTDSGAΞAKNWKKIAGIWYYFNKENQMEIGWIQDKΞQWYYLDV DGSMKTGWLQYMGQ YYFAPSGE
/ SP045 nucleotide (SEQ ID NO:71)
CTTGGGTGTAACCCATATCCAGCTCCTTCCAGTCTTGTCTTACTACTTTGTCAATGAATTGAAAAACCA TGAACGCTTGTCTGACTACGCTTCAAGCAACAGCAACTACAACTGGGGATATGACCCTCAAAACTACTT CTCCTTGACTGGTATGTACTCAAGCGATCCTAAGAATCCAGAAAAACGAATCGCAGAATTTAAAAACCT CATCAACGAAATCCACAAACGTGGTATGGGAGCTATCCTAGATGTCGTTTATAACCACACAGCCAAAGT CGATCTCTTTGAAGATTTGGAACCAAACTACTACCACTTTATGGATGCCGATGGCACACCTCGAACTAG CTTTGGTGGTGGACGCTTGGGGACAACCCACCATATGACCAAACGGCTCCTAATTGACTCTATCAAATA CCTAGTTGATACCTACAAAGTGGATGGCTTCCGTTTCGATATGATGGGAGACCATGACGCCGCTTCTAT CGAAGAAGCTTACAAGGCTGCACGCGCCCTCAATCCAAACCTCATCATGCTTGGTGAAGGTTGGAGAAC CTATGCCGGTGATGAAAACATGCCTACTAAAGCTGCTGACCAAGATTGGATGAAACATACCGATACTGT CGCTGTCTTTTCAGATGACATCCGTAACAACCTCAAATCTGGTTATCCAAACGAAGGTCAACCTGCCTT TATCACAGGTGGCAAGCGTGATGTCAACACCATCTTTAAAAATCTCATTGCTCAACCAACTAACTTTGA AGCTGACAGCCCTGGAGATGTCATCCAATACATCGCAGCCCATGATAACTTGACCCTCTTTGACATCAT TGCCCAGTCTATCAAAAAAGACCCAAGCAAGGCTGAGAACTATGCTGAAATCCACCGTCGTTTACGACT TGGAAATCTCATGGTCTTGACAGCTCAAGGAACTCCATTTATCCACTCCGGTCAGGAATATGGACGTAC TAAACAATTCCGTGACCCAGCCTACAAGACTCCAGTAGCAGAGGATAAGGTTCCAAACAAATCTCACTT GTTGCGTGATAAGGACGGCAACCCATTTGACTATCCTTACTTCATCCATGACTCTTACGATTCTAGTGA TGCAGTCAACAAGTTTGACTGGACTAAGGCTACAGATGGTAAAGCTTATCCTGAAAATGTCAAGAGCCG TGACTATATGAAAGGTTTGATTGCCCTTCGTCAATCTACAGATGCCTTCCGACTTAAGAGTCTTCAAGA TATCAAAGACCGTGTCCACCTCATCACTGTCCCAGGCCAAAATGGTGTGGAAAAAGAGGATGTAGTGAT TGGCTACCAAATCACTGCTCCAAACGGCGATATCTACGCAGTCTTTGTCAATGCGGATGAAAAAGCTCG CGAATTTAATTTGGGAACTGCCTTTGCACATCTAAGAAATGCGGAAGTTTTGGCAGATGAAAACCAAGC AGGACCAGTCGGAATTGCCAACCCGAAAGGACTTGAATGGACTGAAAAAGGCTTGAAATTGAATGCCCT TACAGCTACTGTTCTTCGAGTCTCTCAAAATGGAACTAGCCATGAGTCAACTGCAGAAGAGAAACCAGA CTCAACCCCTTCCAAGCCTGAACATCAAAATGAAGCTTCTCACCCTGCACATCAAGACCCAGCTCCAGA AGCTAGACCTGATTCTACTAAACCAGATGCCAAAGTAGCTGATGCGGAAAATAAACCTAGCCAAGCTAC AGCTGATTCACAAGCTGAACAACCAGCACAAGAAGCACAAGCATCATCTGTAAAAGAAGCGGTTCGAAA CGAATCGGTAGAAAACTCTAGCAAGGAAAATATACCTGCAACCCCAGATAAACAAGCTGAA
SP045 nucleotide (SEQ ID NO:72)
LGVTHIQLLPVLSYYFVNELKNHERLSDYASSNSNYN GYDPQNYFSLTGMYSSDPKNPEKRIAEFKNL INEIHKRGMGAILDWYNHTAKVΌLFEDLEPNYYHF DADGTPRTSFGGGRLGTTHHMTKRLLIDSIKY LVDTYKVDGFRFDMMGDHDAAS I EEAYKAARA NPNLI LGEGWRTYAGDENMPTKAADQD MKHTDTV Table 1 64
AVFSDDIRNNLKSGYPNEGQPAFITGGKRDVNTIF NLIAQPTNFEADSPGDVIQYIAAHDN TLFDII AQSIKKDPSKAENYAEIHRRLRLGNLMVLTAQGTPFIHSGQEYGRTKQFRDPAYKTPVAEDKVPNKSHL LRD DGNPFDYPYFIHDSYDSSDAVNKFD TKATDGKAYPENVKSRDYMKGLIALRQSTDAFRLKSLQD IKDRVH ITVPGQNGVEKEDWIGYQITAPNGDIYAVFV ADEKAREFNLGTAFAHLRNAEVLADENQA GPVGIANPKGLE TEKGLKLNALTATVLRVSQNGTSHESTAEEKPDSTPSKPEHQNEASHPAHQDPAPE ARPDSTKPDAKVADAENKPSQATADSQAEQPAQEAQASSVKEAVRNESVENΞSKENIPATPDKQAΞ
SP046 nucleotide (SEQ ID NO:73)
TAGTGATGGTACTTGGCAAGGAAAACAGTATCTGAAAGAAGATGGCAGTCAAGCAGCAAATGAGTGGGT TTTNGATACTCATTATCAATCTTGGTTCTATATAAAAGCAGATGCTAACTATGCTGAAAATGAATGGCT AAAGCAAGGTGACGACTATTTTTACCTCAAATCTGGTGGCTATATGGCCAAATCAGAATGGGTAGAAGA CAAGGGAGCCTTTTATTATCTTGACCAAGATGGAAAGATGAAAAGAAATGCTTGGGTAGGAACTTCCTA TGTTGGTGCAACAGGTGCCAAAGTAATAGAAGACTGGGTCTATGATTCTCAATACGATGCTTGGTTTTA TATCAAAGCAGATGGACAGCACGCAGAGAAAGAATGGCTCCAAATTAAAGGGAAGGACTATTATTTCAA ATCCGGTGGTTATCTACTGACAAGTCAGTGGATTAATCAAGCTTATGTGAATGCTAGTGGTGCCAAAGT ACAGCAAGGTTGGCTTTTTGACAAACAATACCAATCTTGGTTTTACATCAAAGAAAATGGAAACTATGC TGATAAAGAATGGATTTTCGAGAATGGTCACTATTATTATCTAAAATCCGGTGGCTACATGGCAGCCAA TGAATGGATTTGGGATAAGGAATCTTGGTTTTATCTCAAATTTGATGGGAAAATGGCTGAAAAAGAATG GGTCTACGATTCTCATAGTCAAGCTTGGTACTACTTCAAATCCGGTGGTTACATGACAGCCAATGAATG GATTTGGGATAAGGAATCTTGGTTTTACCTCAAATCTGATGGGAAAATAGCTGAAAAAGAATGGGTCTA CGATTCTCATAGTCAAGCTTGGTACTACTTCAAATCTGGTGGCTACATGGCGAAAAATGAGACAGTAGA TGGTTATCAGCTTGGAAGCGATGGTAAATGGCTTGGAGGAAAAACTACAAATGAAAATGCTGCTTACTA TCAAGTAGTGCCTGTTACAGCCAATGTTTATGATTCAGATGGTGAAAAGCTTTCCTATATATCGCAAGG TAGTGTCGTATGGCTAGATAAGGATAGAAAAAGTGATGACAAGCGCTTGGCTATTACTATTTCTGGTTT GTCAGGCTATATGAAAACAGAAGATTTACAAGCGCTAGATGCTAGTAAGGACTTTATCCCTTATTATGA GAGTGATGGCCACCGTTTTTATCACTATGTGGCTCAGAATGCTAGTATCCCAGTAGCTTCTCATCTTTC TGATATGGAAGTAGGCAAGAAATATTATTCGGCAGATGGCCTGCATTTTGATGGTTTTAAGCTTGAGAA TCCCTTCCTTTTCAAAGATTTAACAGAGGCTACAAACTACAGTGCTGAAGAATTGGATAAGGTATTTAG TTTGCTAAACATTAACAATAGCCTTTTGGAGAACAAGGGCGCTACTTTTAAGGAAGCCGAAGAACATTA CCATATCAATGCTCTTTATCTCCTTGCCCATAGTGCCCTAGAAAGTAACTGGGGAAGAAGTAAAATTGC CAAAGATAAGAATAATTTCTTTGGCATTACAGCCTATGATACGACCCCTTACCTTTCTGCTAAGACATT TGATGATGTGGATAAGGGAATTTTAGGTGCAACCAAGTGGATTAAGGAAAATTATATCGATAGGGGAAG AACTTTCCTTGGAAACAAGGCTTCTGGTATGAATGTGGAATATGCTTCAGACCCTTATTGGGGCGAAAA AATTGCTAGTGTGATGATGAAAATCAATGAGAAGCTAGGTGGCAAAGAT
SP046 amino acid (SEQ ID NO:74)
SIX-T QGKQYLKEDGSQAANEWVXDTHYQSWFYIKADANYAENE LKQGDDYFYLKΞGGYMAKSEWVED KGAFYYLDQDGKMKRNAWVGTSYVGATGAKVIEDWVYDSQYDAWFYIKADGQHAEKEWLQIKGKDYYFK SGGYLLTSQWINQAYV ASGAKVQQGWLFDKQYQS FYIKENGNYADKE IFENGHYYYLKSGGYMAAN E I DKESWFYLKFDGKMAEKEWVΎDSHSQAWYYFKSGGYMTANE IWDKESWFYLKΞDGKIAEKE VΎ DSHSQAWYYFKSGGYMAKNETVDGYQLGΞDGKWLGGKTTNENAAYYQVVPVTANVYDSDGEKLSYISQG SWWLDKDRKSDDKRLAITISGLSGYMKTEDLQALDASKDFIPYYESDGHRFYHYVAQNASIPVASHLS DMEVGKKYYSADGLHFDGFKLENPFLFKDLTEATNYSAEELDKVFSLLNI NS LENKGATFKEAEEHY HINALYLLAHSALESN GRSKIAKDKNNFFGITAYDTTPYLSAKTFDDVDKGI GATKWIKENYIDRGR TFLGNKASGMNVEYASDPYWGEKIASVMMKINEKLGGKD
SP048 nucleotide (SEQ ID NO:75)
TGGGATTCAATATGTCAGAGATGATACTAGAGATAAAGAAGAGGGAATAGAGTATGATGACGCTGACAA TGGGGATATTATTGTAAAAGTAGCGACTAAACCTAAGGTAGTAACCAAGAAAATTTCAAGTACGCGAAT TCGTTATGAAAAAGATGAAACAAAAGACCGTAGTGAAAATCCTGTTACAATTGATGGAGAGGATGGCTA TGTAACTACGACAAGGACCTACGATGTTAATCCAGAGACTGGTTATGTTACCGAACAGGTTACTGTTGA TAGAAAAGAAGCCACGGATACAGTTATCAAAGTTCCAGCTAAAAGCAAGGTTGAAGAAGTTCTTGTTCC ATTTGCTACTAAATATGAAGCAGACAATGACCTTTCTGCAGGACAGGAGCAAGAGATTACTCTAGGAAA GAATGGGAAAACAGTTACAACGATAACTTATAATGTAGATGGAAAGAGTGGACAAGTAACTGAGAGTAC TTTAAGTCAAAAAAAAGACTCtCAAACAAGAGTTGTTAAAAAAAGaACCArkCCCCAAGTTCTTGTCCA AGAAATTCCAATCGAAACAGAATATCTCGATGGCCCaACTCTTGATAAAaGTCAAGAAGTAGAAGAAGT AGGAGAAATTGGTAAATTACTCTTACTACAATCTATACTGGTAGATGAACGTGATGGAACAATTGAAGA AACTACTTCTCGTCAAATTACTAAAGAGATGGTAAAAAGACGTATAAGGAGAGGGACGAGAGAACCTGA Table 1 65
AAAAGTTGTTGTTCCTGAGCAATCATCTATTCCTTCGTATCCTGTATCTGTTACATCTAACCAAGGAAC AGATGTAGCAGTAGAACCAGCTAAAGCAGTTGCTCCAACAACAGACTGGAAACAAGAAAATGGTATGTG GTATTTTTATAATACTGATGGTTCCATGGCAACAGGTTGGGTACAAGTTAATAGTTCATGGTACTACCT CAACAGCAACGGTTCTATGAAAGTCAATCAATGGTTCCAAGTTGGTGGTAAATGGTATTATGTAAATAC ATCGGGTGAGTTAGCGGTCAATACAAGTATAGATGGCTATAGAGTCAATGATAATGGTGAATGGGTGCG T
SP048 amino acid (SEQ ID NO:76)
GIQYVRDDTRDKEEGIEYDDADNGDIIVKVATKPKWTKKISSTRIRYEKDETKDRSENPVTIDGEDGY VTTTRTYDVNPETGYVTEQVTVDRKEATDTVIKVPAKSKVEEVLVPFATKYEADNDLSAGQEQEITLGK NGKTVTTITYNVDGKSGQVTESTLSQKKDSQTRWKKRTXPQVLVQEIPIETΞYLDGPTLDKΞQEVEEV GEIGK LL QSILVDERDGTIEETTSRQITKEMVKRRIRRGTREPEKVWPEQSSIPSYPVSVTΞNQGT DVAVEPAKAVAPTTDWKQENGMW.'FY TDGSMATG VQVNSS YYLNSNGS KV Q FQVGGKWYYVNT SGELAVNTSIDGYRVNDNGE VR
SP049 nucleotide (SEQ ID NO:77)
GGATAATAGAGAAGCATTAAAAACCTTTATGACGGGTGAAAATTTTTATCTCCAACATTATCTAGGAGC ACATAGGGAAGAACTAAATGGAGAGCATGGCTATACCTTCCGTGTTTGGGCACCTAATGCTCAGGCTGT TCACTTGGTTGGTGATTTTACCAACTGGATTGAAAATCAGATTCCAATGGTAAGAAATGATTTTGGGGT CTGGGAAGTCTTTACCAATATGGCTCAAGAAGGGCATATTTACAAATATCATGTCACACGTCAAAATGG TCATCAACTGATGAAGATTGACCCTTTTGCTGTCAGGTATGAGGCTCGTCCAGGAACAGGGGCAATCGT AACAGAGCTTCCTGAGAAGAAATGGAAGGATGGACTTTGGCTGGCACGAAGAAAACGTTGGGGCTTTGA AGAGCGTCCTGTCAATATTTATGAAGTTCACGCTGGATCATGGAAAAGAAATTCTGATGGCAGTCCTTA TAGTTTTGCCCAGCTCAAGGATGAACTCATTCCTTATCTCGTTGAAATGAACTATACTCATATTGAGTT TATGCCCTTGATGTCGCATCCTTTGGGCTTGAGTTGGGGGTATCAGCTTATGGGTTACTTCGCTTTAGA GCATGCTTATGGCCGACCAGAGGAGTTTCAAGATTTTGTC
SP049 amino acid (SEQ ID NO:78)
DNREALKTFMTGENFYLQHYLGAHREELNGEHGYTFRVWAPNAQAVHLVGDFTN IENQIPMVRNDFGV WEVFTNMAQEGHIYKYHVTRQNGHQLMKIDPFAVRYEARPGTGAIVTELPEKKWKDGL LARRKR GFE ERPVNIYEVHAGSWKRNΞDGSPYSFAQLKDELIPYLVEMNYTHIEFMPLMSHPLGLSWGYQLMGYFALE HAYGRPEEFQDFV
SP050 nucleotide (SEQ ID NO:79)
AGATTTTGTCGAGGAGTGTCATACCCATAATATTGGGGTTATTGTGGACTGGGTACCAGNTCACTTTAC CATCAACGATGATGCCTTAGCCTATTATGATGGGACACCGACTTTTGAATACCAAGACCATAATAAGGC TCATAACCATGGTTGGGGTGCCCTTAATTTTGACCTTGGAAAAAATGAAGTCCAGTCCTTCTTAATTTC TTGCATTAAGCATTGGATTGATGTCTATCATTTGGATGGTATTCGTGTGGATGCTGTTAGCAACATGCT CTATTTGGACTATGATGATGCTCCATGGACACCTAATAAAGATGGCGGAAATCTCAACTATGAAGGTTA TTATTTCCTTCAGCGCTTGAATGAGGTTATTAAGTTAGAATATCCAGATGTGATGATGATTGCAGAAGA AAGTTCGTCTGCGATCAAGATTACGGGAATGAAAGAGATTGGTGGTCTAGGATTTGACTACAAATGGAA CATGGGCTGGATGAATGATATCCTCCGTTTCTACGAAGAAGATCCGATCTATCGTAAATATGACTTTAA CCTGGTGACTTTCAGCTTTATGTATGTTTNCAAGGAGAATTATCTCTTGCCATTCTCGCACGATGAAGT GGTTCATGGCAAGAAGAGTATGATGCATAAGATGTGGGGAGATCGTTACAATCAATTCGCAGGCTTGCG CAATCTCTATACGTACCAAATTTGTCACCCTGGTAAGAAATTGCTCTTCATGGGTAGCGAATACGGTCA ATTCCTAGAATGGAAATCTGAAGAACAGTTGGAATGGTCTAACCTAGAAGACCCAATGAATGCTAAGAT GAAGTATTTCGCTTCTCAGCTAAACCAGTTTTACAAAGATCATCGCTGTCTGTGGGAAATTGATACCAG CTATGATGGTATTGAAATCATTGATGCGGATAATCGAGACCAGAGTGTTCTTTCCTTTATTCGTAAGGG TAAAAAGGGA
SP050 amino acid (SEQ ID NO:80)
DFVEECHTHNIGVIVDWVPXHFTINDDA AYYDGTPTFEYQDHNKAHNHGWGALNFDLGKNEVQΞFLIS CIKHWIDVYHLDGIRVDAVSNM YLDYDDAP TPNKDGGNLNYEGYYFLQRLNEVIKLEYPDVMMIAEE SSSAIKITGMKEIGGLGFDYKWNMG MNDILRFYEEDPIYRKYDFNLVTFSFMYVXKE Y LPFSHDEV VHGKKS MHKMWGDRYNQFAG R LYTYQICHPGKKLLFMGSEYGQFLEWKSEEQLEWSNLEDPMNAK KYFASQLNQFYKDHRCLWEIDTSYDGIEIIDADNRDQSVLSFIRKGKKG
SP051 nucleotide (SEQ ID NO: 81) Table 1 66
ATCTGTAGTTTATGCGGATGAAACACTTATTACTCATACTGCTGAGAAACCTAAAGAGGAAAAAATGAT AGTAGAAGAAAAGGCTGATAAAGCTTTGGAAACTAAAAATATAGTTGAAAGGACAGAACAAAGTGAACC TAGTTCAACTGAGGCTATTGCATCTGAGNAGAAAGAAGATGAAGCCGTAACTCCAAAAGAGGAAAAAGT GTCTGCTAAACCGGAAGAAAAAGCTCCAAGGATAGAATCACAAGCTTCAAATCAAGAAAAACCGCTCAA GGAAGATGCTAAAGCTGTAACAAATGAAGAAGTGAATCAAATGATTGAAGACAGGAAAGTGGATTTTAA TCAAAATTGGTACTTTAAACTCAATGCAAATTCTAAGGAAGCCATTAAACCTGATGCAGACGTATCTAC GTGGAAAAAATTAGATTTACCGTATGACTGGAGTATCTTTAACGATTTCGATCATGAATCTCCTGCACA AAATGAAGGTGGACAGCTCAACGGTGGGGAAGCTTGGTATCGCAAGACTTTCAAACTAGATGAAAAAGA CCTCAAGAAAAATGTTCGCCTTACTTTTGATGGCGTCTACATGGATTCTCAAGTTTATGTCAATGGTCA GTTAGTGGGGCATTATCCAAATGGTTATAACCAGTTCTCATATGATATCACCAAATACCTTCAAAAAGA TGGTCGTGAGAATGTGATTGCTGTCCATGCAGTCAACAAACAGCCAAGTAGCCGTTGGTATTCAGGAAG TGGTATCTATCGTGATGTGACTTTACAAGTGACAGATAAGGTGCATGTTGAGAAAAATGGGACAACTAT TTTAACACCAAAACTTGAAGAACAACAACATGGCAAGGTTGAAACTCATGTGACCAGCAAAATCGTCAA TACGGACGACAAAGACCATGAACTTGTAGCCGAATATCAAATCGTTGAACGAGGTGGTCATGCTGTAAC AGGCTTAGTTCGTACAGCGAGTCGTACCTTAAAAGCACATGAATCAACAAGCCTAGATGCGATTTTAGA AGTTGAAAGACCAAAACTCTGGACTGTTTTAAATGACAAACCTGCCTTGTACGAATTGATTACGCGTGT TTACCGTGACGGTCAATTGGTTGATGCTAAGAAGGATTTGTTTGGTTACCGTTACTATCACTGGACTCC AAATGAAGGTTTCTCTTTGAATGGTGAACGTATTAAATTCCATGGAGTATCCTTGCACCACGACCATGG GGCGCTTGGAGCAGAAGAAAACTATAAAGCAGAATATCGCCGTCTCAAACAAATGAAGGAGATGGGAGT TAACTCCATCCGTACAACCCACAACCCTGCTAGTGAGCAAACCTTGCAAATCGCAGCAGAACTAGGTTT ACTCGTTCAGGAAGAGGCCTTTGATACGTGGTATGGTGGCAAGAAACCTTATGACTATGGACGTTTCTT TGAAAAAGATGCCACTCACCCAGAAGCTCGAAAAGGTGAAAAATGGTCTGATTTTGACCTACGTACCAT GGTCGAAAGAGGCAAAAACAACCCTGCTATCTTCATGTGGTCAATTGGTAATGAAATAGGTGAAGCTAA TGGTGATGCCCACTCTTTAGCAACTGTTAAACGTTTGGTTAAGGTTATCAAGGATGTTGATAAGACTCG CTATGTTACCATGGGAGCAGATAAATTCCGTTTCGGTAATGGTAGCGGAGGGCATGAGAAAATTGCTGA TGAACTCGATGCTGTTGGATTTAACTATTCTGAAGATAATTACAAAGCCCTTAGAGCTAAGCATCCAAA ATGGTTGATTTATGGATCAGAAACATCTTCAGCTACCCGTACACGTGGAAGTTACTATCGCCCTGAACG TGAATTGAAACATAGCAATGGACCTGAGCGTAATTATGAACAGTCAGATTATGGAAATGATCGTGTGGG TTGGGGGAAAACAGCAACCGCTTCATGGACTTTTGACCGTGACAACGCTGGCTATGCTGGACAGTTTAT CTGGACAGGTACGGACTATATTGGTGAACCTACACCATGGCACAACCAAAATCAAACTCCTGTTAAGAG CTCTTACTTTGGTATCGTAGATACAGCCGGCATTCCAAAACATGACTTCTATCTCTACCAAAGC
SP051 amino acid (SEQ ID NO:82)
SWYADETLITHTAEKPKEEK IVEEKADKALET NIVΞRTEQSEPSSTEAIASEXKEDEAVTPKEEKV SAKPEEKAPRIESQASNQEKPL ΞDAKAVTNEEVNQMIEDRKVDFNQN YFK NANΞKEAIKPDADVST WKKLDLPYD SIFNDFDHESPAQNEGGQLNGGEAWYRKTFKLDEKDLKKIWRLTFDGVYMDSQVYVNGQ LVGHYPNGYNQFSYDITKYLQKDGRE VIAVHAVNKQPSSRWYSGΞGIYRDVTLQVTDKVHVEKNGTTI LTPK EEQQHGKVETHVTSKIVNTDDKDHELVAEYQIVERGGHAVTGLVRTAΞRTLKAHEΞTS DAILE VERPKL TVLNDKPALYELITRVYRDGQLVDAKKDLFGYRYYHWTPNEGFS GERIKFHGVSLHHDHG A GAEENYKAEYRRLKQMKEMGV ΞIRTTHNPASEQTLQIAAELGLLVQEEAFDTWYGGKKPYDYGRFF EKDATHPEARKGEK SDFDLRTMVERGKNNPAIFM SIGNEIGEANGDAHSLATVKRLVKVIKDVDKTR YVTMGADKFRFGNGSGGHEKIADELDAVGFNYSED YKALRAKHPKWLIYGSΞTSSATRTRGSYYRPER ELKHSNGPER YEQSDYGNDRVGWGKTATASWTFDRDNAGYAGQFIWTGTDYIGEPTPWH QNQTPVKS SYFGIVDTAGIPKHDFYLYQS
SP052 nucleotide (SEQ ID NO: 83)
TTACTTTGGTATCGTAGATACAGCCGGCATTCCAAAACATGACTTCTATCTCTACCAAAGCCAATGGGT TTCTGTTAAGAAGAAACCGATGGTACACCTTCTTCCTCACTGGAACTGGGAAAACAAAGAATTAGCATC CAAAGTAGCTGACTCAGAAGGTAAGATTCCAGTTCGTGCTTATTCGAATGCTTCTAGTGTAGAATTGTT CTTGAATGGAAAATCTCTTGGTCTTAAGACTTTCAATAAAAAACAAACCAGCGATGGGCGGACTTACCA AGAAGGTGCAAATGCTAATGAACTTTATCTTGAATGGAAAGTTGCCTATCAACCAGGTACCTTGGAAGC AATTGCTCGTGATGAATCTGGCAAGGAAATTGCTCGAGATAAGATTACGACTGCTGGTAAGCCAGCGGC AGTTCGTCTTATTAAGGAAGACCATGCGATTGCAGCAGATGGAAAAGACTTGACTTACATCTACTATGA AATTGTTGACAGCCAGGGGAATGTGGTTCCAACTGCTAATAATCTGGTTCGCTTCCAATTGCATGGCCA AGGTCAACTGGTCGGTGTAGATAACGGAGAACAAGCCAGCCGTGAACGCTATAAGGCGCAAGCAGATGG TTCTTGGATTCGTAAAGCATTTAATGGTAAAGGTGTTGCCATTGTCAAATCAACTGAACAAGCAGGGAA ATTCACCCTGACTGCCCACTCTGATCTCTTGAAATCGAACCAAGTCACTGTCTTTACTGGTAAGAAAGA AGGACAAGAGAAGACTGTTTTGGGGACAGAAGTGCCAAAAGTACAGACCATTATTGGAGAGGCACCTGA Table 1 67
AATGCCTACCACTGTTCCGTTTGTATACAGTGATGGTAGCCGTGCAGAACGTCCTGTAACCTGGTCTTC AGTAGATGTGAGCAAGCCTGGTATTGTAACGGTGAAAGGTATGGCTGACGGACGAGAAGTAGAAGCTCG TGTAGAAGTGATTGCTCTTAAATCAGAGCTACCAGTTGTGAAACGTATTGCTCCAAATACTGACTTGAA TTCTGTAGACAAATCTGTTTCCTATGTTTTGATTGATGGAAGTGTTGAAGAGTATGAAGTGGACAAGTG GGAGATTGCCGAAGAAGATAAAGCTAAGTTAGCAATTCCAGGTTCTCGTATTCAAGCGACCGGTTATTT AGAAGGTCAACCAATTCATGCAACCCTTGTGGTAGAAGAAGGCAATCCTGCGGCACCTGCAGTACCAAC TGTAACGGTTGGTGGTGAGGCAGTAACAGGTCTTACTAGTCAAAAACCAATGCAATACCGCACTCTTGC TTATGGAGCTAAGTTGCCAGAAGTCACAGCAAGTGCTAAAAATGCAGCTGTTACAGTTCTTCAAGCAAG CGCAGCAAACGGCATGCGTGCGAGCATCTTTATTCAGCCTAAAGATGGTGGCCCTCTTCAAACCTATGC AATTCAATTCCTTGAAGAAGCGCCAAAAATTGCTCACTTGAGCTTGCAAGTGGAAAAAGCTGACAGTCT CAAAGAAGACCAAACTGTCAAATTGTCGGTTCGAGCTCACTATCAAGATGGAACGCAAGCTGTATTACC AGCTGATAAAGTAACCTTCTCTACAAGTGGTGAAGGGGAAGTCGCAATTCGTAAAGGAATGCTTGAGTT GCATAAGCCAGGAGCAGTCACTCTGAACGCTGAATATGAGGGAGCTAAAGACCAAGTTGAACTCACTAT CCAAGCCAATACTGAGAAGAAGATTGCGCAATCCATCCGTCCTGTAAATGTAGTGACAGATTTGCATCA GGAACCAAGTCTTCCAGCAACAGTAACAGTTGAGTATGACAAAGGTTTCCCTAAAACTCATAAAGTCAC TTGGCAAGCTATTCCGAAAGAAAAACTAGACTCCTATCAAACATTTGAAGTACTAGGTAAAGTTGAAGG AATTGACCTTGAAGCGCGTGCAAAAGTCTCTGTAGAAGGTATCGTTTCAGTTGAAGAAGTCAGTGTGAC AACTCCAATCGCAGAAGCACCACAATTACCAGAAAGTGTTCGGACATATGATTCAAATGGTCACGTTTC ATCAGCTAAGGTTGCATGGGATGCGATTCGTCCAGAGCAATACGCTAAGGAAGGTGTCTTTACAGTTAA TGGTCGCTTAGAAGGTACGCAATTAACA
SP052 amino acid (SEQ ID NO: 84)
YFGIVϋTAGIPKHDFYLYQSQWVSVKKKPMVHLLPH NWENKELASKVADSEGKIPVRAYSNASSVELF LNGKSLGLKTFNKKQTSDGRTYQEGANANELYLEWKVAYQPGTLEAIARDESGKEIARDKITTAGKPAA VRLIKEDHAIAADGKDLTYIYYEIVDSQGNWPTANNLVRFQLHGQGQLVGVDNGEQASRERYKAQADG S IRKAFNGKGVAIVKSTEQAGKFTLTAHSDLLKSNQVTVFTGKKEGQEKTVLGTEVPKVQTIIGEAPE MPTTVPFVYSDGSRAERPVT SSVDVSKPGIVTVKGMADGREVEARVEVIALKSELPVVKRIAPNTDLN SVDKSVSYVLIDGSVEEYEVDK EIAEEDKAKLAIPGΞRIQATGYLEGQPIHATLWEEGNPAAPAVPT VTVGGEAVTGLTΞQKPMQYRTLAYGAKLPEVTASAKNAAVTV QASAANG RASIFIQPKDGGP QTYA IQFLEEAPKIAHLSLQVEKADSLKEDQTVKLSVRAHYQDGTQAVLPADKVTFΞTSGEGEVAIRKGMLEL HKPGAVTLNAEYΞGAKDQVELTIQANTEKKIAQSIRPVNWTDLHQEPSLPATVTVEYDKGFPKTHKVT WQAIPKEKLDΞYQTFEVLGKVEGIDLEARAKVSVEGIVSVEEVSVTTPIAEAPQLPESVRTYDSNGHVS SAKVA DAIRPΞQYAKEGVFTV GRLEGTQLT
SP053 nucleotide (SEQ ID NO: 85)
AGCTAAGGTTGCATGGGATGCGATTCGTCCAGAGCAATACGCTAAGGAAGGTGTCTTTACAGTTAATGG TCGCTTAGAAGGTACGCAATTAACAACTAAACTTCATGTTCGCGTATCTGCTCAAACTGAGCAAGGTGC AAACATTTCTGACCAATGGACCGGTTCAGAATTGCCACTTGCCTTTGCTTCAGACTCAAATCCAAGCGA CCCAGTTTCAAATGTTAATGACAAGCTCATTTCCTACAATAACCAACCAGCCAATCGTTGGACAAACTG GAATCGTACTAATCCAGAAGCTTCAGTCGGTGTTCTGTTTGGAGATTCAGGTATCTTGAGCAAACGCTC CGTTGATAATCTAAGTGTCGGATTCCATGAAGACCATGGAGTTGGTGTACCGAAGTCTTATGTGATTGA GTATTATGTTGGTAAGACTGTCCCAACAGCTCCTAAAAACCCTAGTTTTGTTGGTAATGAGGACCATGT CTTTAATGATTCTGCCAACTGGAAACCAGTTACTAATCTAAAAGCCCCTGCTCAACTCAAGGCTGGAGA AATGAACCACTTTAGCTTTGATAAAGTTGAAACCTATGCTGTTCGTATTCGCATGGTTAAAGCAGATAA CAAGCGTGGAACGTCTATCACAGAGGTACAAATCTTTGCGAAACAAGTTGCGGCAGCCAAGCAAGGACA AACAAGAATCCAAGTTGACGGCAAAGACTTAGCAAACTTCAACCCTGATTTGACAGACTACTACCTTGA GTCTGTAGATGGAAAAGTTCCGGCAGTCACAGCAAGTGTTAGCAACAATGGTCTCGCTACCGTCGTTCC AAGCGTTCGTGAAGGTGAGCCAGTTCGTGTCATCGCGAAAGCTGAAAATGGCGACATCTTAGGAGAATA CCGTCTGCACTTCACTAAGGATAAGAGCTTACTTTCTCATAAACCAGTTGCTGCGGTTAAACAAGCTCG CTTGCTACAAGTAGGTCAAGCACTTGAATTGCCGACTAAGGTTCCAGTTTACTTCACAGGTAAAGACGG CTACGAAACAAAAGACCTGACAGTTGAATGGGAAGAAGTTCCAGCGGAAAATCTGACAAAAGCAGGTCA ATTTACTGTTCGAGGCCGTGTCCTTGGTAGTAACCTTGTTGCTGAGATCACTGTACGAGTGACAGACAA ACTTGGTGAGACTCTTTCAGATAACCCTAACTATGATGAAAACAGTAACCAGGCCTTTGCTTCAGCAAC CAATGATATTGACAAAAACTCTCATGACCGCGTTGACTATCTCAATGACGGAGATCATTCAGAAAATCG TCGTTGGACAAACTGGTCACCAACACCATCTTCTAATCCAGAAGTATCAGCGGGTGTGATTTTCCGTGA AAATGGTAAGATTGTAGAACGGACTGTTACACAAGGAAAAGTTCAGTTCTTTGCAGATAGTGGTACGGA TGCACCATCTAAACTCGTTTTAGAACGCTATGTCGGTCCAGAGTTTGAAGTGCCAACCTACTATTCAAA CTACCAAGCCTACGACGCAGACCATCCATTCAACAATCCAGAAAATTGGGAAGCTGTTCCTTATCGTGC Table 1 68
GGATAAAGACATTGCAGCTGGTGATGAAATCAACGTAACATTTAAAGCTATCAAAGCCAAAGCTATGAG ATGGCGTATGGAGCGTAAAGCAGATAAGAGCGGTGTTGCGATGATTGAGATGACCTTCCTTGCACCAAG TGAATTGCCTCAAGAAAGCACTCAATCAAAGATTCTTGTAGATGGAAAAGAACTTGCTGATTTCGCTGA AAATCGTCAAGACTATCAAATTACCTATAAAGGTCAACGGCCAAAAGTCTCAGTTGAAGAAAACAATCA AGTAGCTTCAACTGTGGTAGATAGTGGAGAAGATAGCTTTCCAGTACTTGTTCGCCTCGTTTCAGAAAG TGGAAAACAAGTCAAGGAATACCGTATCCACTTGACTAAGGAAAAACCAGTTTCTGAGAAGACAGTTGC TGCTGTACAAGAAGATCTTCCAAAAATCGAATTTGTTGAAAAAGATTTGGCATACAAGACAGTTGAGAA AAAAGATTCAACACTGTATCTAGGTGAAACTCGTGTAGAACAAGAAGGAAAAGTTGGAAAAGAACGTAT CTTTACAGCGATTAATCCTGATGGAAGTAAGGAAGAAAAACTCCGTGAAGTGGTAGAAGTTCCGACAGA CCGCATCGTCTTGGTTGGAACCAAACCAGTAGCTCAAGAAGCTAAAAAACCACAAGTGTCAGAAAAAGC AGATACAAAACCAATTGATTCAAGTGAAGCTAGTCAAACTAATAAAGCCCAG
SP053 amino acid (SEQ ID NO: 86)
A VAWDAIRPEQYAKEGVFTVNGRLEGTQLTTKLHVRVSAQTEQGANISDQ TGSELPLAFASDSNPSD PVSNVNDKLISYNNQPANRWTIΛTORTNPEASVGVLFGDSGILSKRSVDNLSVGFHEDHGVGVPKSYVIE YYVGKTVPTAPKMPSFVGNEDHVFNDSANWKPVTNLKAPAQLKAGEMNHFSFDKVETYAVRIRMVKADN KRGTSITΞVQIFAKQVAAAKQGQTRIQVDGKDLANFNPDLTDYYLESVDGKVPAVTASVSNNGLATλΛ/P SVREGEPVRVIAKAENGDI GEYRLHFTKDKS LSHKPVAAVKQARLLQVGQALELPTKVPVYFTGKDG YETKDLTVE EEVPAENLTKAGQFTVRGRVLGSNLVAEITVRVTDKLGETLSDNPNYDENSNQAFASAT NDIDKNSHDRVDYLNDGDHSENRR TNWSPTPSSNPEVSAGVIFRENGKIVERTVTQGKVQFFADSGTD APSKLV ERYVGPEFEVPTYYΞNYQAYDADHPFNNPENWEAVPYRADKDIAAGDEINVTFKAIKAKAMR RMERKADKSGVAMIEMTFLAPSELPQEΞTQSKILVDGKELADFAENRQDYQITYKGQRPKVSVEENNQ VASTVVDSGEDSFPVLVRLVSESGKQVKEYRIHLTKEKPVSEKTVAAVQEDLPKIEFVEKDLAYKTVEK KDSTLY GETRVΞQEGKVGKERIFTAINPDGSKEEKLREWEVPTDRIVLVGTKPVAQEAKKPQVSEKA DTKPIDSSEASQTNKAQ
SP054 nucleotide (SEQ ID NO: 87)
CTATCACTATGTAAATAAAGAGATTATTTCACAAGAAGCTAAAGATTTAATTCAGACAGGAAAGCCTGA CAGGAATGAAGTTGTATATGGTTTGGTGTATCAAAAAGATCAGTTGCCTCAAACAGGGACAGAA
SP054 amino acid (SEQ ID NO: 88)
YHYVNKEIISQEAKDLIQTGKPDRNEWYGLVYQKDQLPQTGTE
SP055 nucleotide (SEQ ID NO:89)
TGAGACTCCTCAATCAATAACAAATCAGGAGCAAGCTAGGACAGAAAACCAAGTAGTAGAGACAGAGGA AGCTCCAAAAGAAGAAGCACCTAAAACAGAAGAAAGTCCAAAGGAAGAACCAAAATCGGAGGTAAAACC TACTGACGACACCCTTCCTAAAGTAGAAGAGGGGAAAGAAGATTCAGCAGAACCAGCTCCAGTTGAAGA AGTAGGTGGAGAAGTTGAGTCAAAACCAGAGGAAAAAGTAGCAGTTAAGCCAGAAAGTCAACCATCAGA CAAACCAGCTGAGGAATCAAAAGTTGAACAAGCAGGTGAACCAGTCGCGCCAAGAGAAGACGAAAAGGC ACCAGTCGAGCCAGAAAAGCAACCAGAAGCTCCTGAAGAAGAGAAGGCTGTAGAGGAAACACCGAAACA AGAAGAGTCAACTCCAGATACCAAGGCTGAAGAAACTGTAGAACCAAAAGAGGAGACTGTTAATCAATC TATTGAACAACCAAAAGTTGAAACGCCTGCTGTAGAAAAACAAACAGAACCAACAGAGGAACCAAAAGT TGAACAAGCAGGTGAACCAGTCGCGCCAAGAGAAGACGAACAGGCACCAACGGCACCAGTTGAGCCAGA AAAGCAACCAGAAGTTCCTGAAGAAGAGAAGGCTGTAGAGGAAACACCGAAACCAGAAGATAAAATAAA GGGTATTGGTACTAAAGAACCAGTTGATAAAAGTGAGTTAAATAATCAAATTGATAAAGCTAGTTCAGT TTCTCCTACTGATTAT
SP055 amino acid (SEQ ID NO:90)
ETPQSITNQEQARTENQWETEEAPKEEAPKTEESPKEEPKSEVKPTDDTLPKVEEGKEDSAEPAPVEE VGGEVESKPEEKVAVKPESQPSDKPAEESKVEQAGEPVAPREDEKAPVEPEKQPEAPEEEKAVEETPKQ EESTPDTKAEETVEPKEETVNQSIEQPKVΞTPAVEKQTEPTEEPKVEQAGEPVAPREDEQAPTAPVEPE KQPEVPEEEKAVEETPKPEDKIKGIGTKEPVDKSELNNQIDKASSVSPTDY
SP056 nucleotide (SEQ ID NO:91)
GGATGCTCAAGAAACTGCGGGAGTTCACTATAAATATGTGGCAGATTCAGAGCTATCATCAGAAGAAAA GAAGCAGCTTGTCTATGATATTCCGACATACGTGGAGAATGATGATGAAACTTATTATCTTGTTTATAA GTTAAATTCTCAAAATCAACTGGCGGAATTGCCAAATACTGGAAGCAAGAATGAGAGGCAA Table 1 69
SP056 amino acid (SEQ ID NO:92)
DAQETAGVHYKYVADSELSSEEKKQLVYDIPTYVENDDETYYLVΎKLNSQNQLAΞLPNTGSKNERQ
SP057 nucleotide (SEQ ID NO:93)
CGACAAAGGTGAGACTGAGGTTCAACCAGAGTCGCCAGATACTGTGGTAAGTGATAAAGGTGAACCAGA GCAGGTAGCACCGCTTCCAGAATATAAGGGTAATATTGAGCAAGTAAAACCTGAAACTCCGGTTGAGAA GACCAAAGAACAAGGTCCAGAAAAAACTGAAGAAGTTCCAGTAAAACCAACAGAAGAAACACCAGTAAA TCCAAATGAAGGTACTACAGAAGGAACCTCAATTCAAGAAGCAGAAAATCCAGTTCAACCTGCAGAAGA ATCAACAACGAATTCAGAGAAAGTATCACCAGATACATCTAGCAAAAATACTGGGGAAGTGTCCAGTAA TCCTAGTGATTCGACAACCTCAGTTGGAGAATCAAATAAACCAGAACATAATGACTCTAAAAATGAAAA TTCAGAAAAAACTGTAGAAGAAGTTCCAGTAAATCCAAATGAAGGCACAGTAGAAGGTACCTCAAATCA AGAAACAGAAAAACCAGTTCAACCTGCAGAAGAAACACAAACAAACTCTGGGAAAATAGCTAACGAAAA ACTGGAGAAGTATCCAATAAACCTAGTGATTCAAAACCACCAGTTGAAGAATCAAATCAACCAGAAAA AAACGGAACTGCAACAAAACCAGAAAATTCAGGTAATACAACATCAGAGAATGGACAAACAGAACCAGA ACCATCAAACGGAAATTCAACTGAGGATGTTTCAACCGAATCAAACACATCCAATTCAAATGGAAACGA AGAAATTAAACAAGAAAATGAACTAGACCCTGATAAAAAGGTAGAAGAACCAGAGAAAACACTTGAATT
AAGAAAT
SP057 amino acid (SEQ ID NO: 94)
DKGETEVQPESPDTWSDKGEPΞQVAPLPEYKGNIΞQVKPETPVEKTKEQGPEKTEEVPVKPTEETPVN PNEGTTEGTSIQEAENPVQPAΞESTTNSEKVSPDTSSKNTGEVSΞNPSDΞTTSVGESNKPEHNDS NEN ΞEKTVEEVPV PNEGTVEGTSNQETΞKPVQPAEETQTNSGKIA ENTGEVSNKPSDSKPPVEESNQPΞK NGTATKPENSGNTTSENGQTEPEPSNGNSTEDVSTESNTSNSNGNEEIKQENELDPDKKVEEPEKTLEL RN
/ SP058 nucleotide (SEQ ID NO:95)
AAATCAATTGGTAGCACAAGATCCAAAAGCACAAGATAGCACTAAACTGACTGCTGAAAAATCAACTGT TAAAGCACCTGCTCAAAGAGTAGATGTAAAAGATATAACτCATTTAACAGATGAAGAAAAAGTTAAGGT TGCTATTTTACAAGCAAATGGTTCAGCATTAGACGGAGCGACAATCAATGTAGCTGGAGATGGTACAGC AACAATCACATTCCCAGATGGTTCAGTAGTGACGATTCTAGGAAAAGATACAGTTCAACAATCTGCGAA AGGTGAATCTGTAACTCAAGAAGCTACACCAGAGTATAAGCTAGAAAATACACCAGGTGGAGATAAGGG AGGCAATACTGGAAGCTCAGATGCTAATGCGAATGAAGGCGGTGGTAGCCAGGCGGGTGGATCAGCTCA CACAGGTTCACAAAACTCAGCTCAATCACAAGCTTCTAAGCAATTAGCTACTGAAAAAGAATCAGCTAA AAATGCCATTGAAAAAGCAGCCAAGGACAAGCAGGATGAAATCAAAGGCGCACCGCTTTCTGATAAAGA AAAAGCAGAACTTTTAGCAAGAGTGGAAGCAGAAAAACAAGCAGCTCTCAAAGAGATTGAAAATGCGAA AACTATGGAAGATGTGAAGGAAGCAGAAACGATTGGAGTGCAAGCCATTGCCATGGTTACAGTTCCTAA GAGACCAGTGGCTCCTAAT
SP058 amino acid (SEQ ID NO:96)
NQLVAQDPKAQDSTKLTAEKSTVKAPAQRVϋVKDITHLTDEEKVKVAILQANGSALDGATINVAGDGTA TITFPDGSWTILGKDTVQQSAKGESVTQEATPΞYKLENTPGGDKGGNTGΞΞDANANEGGGSQAGGSAH TGSQNSAQSQASKQLATEKESAKNAIEKAAKDKQDEIKGAPLSDKEKAELLARVEAEKQAALKEIENAK TMEDVKEAETIGVQAIAMVTVPKRPVAPN
SP059 nucleotide (SEQ ID NO:97)
CAAACAGTCAGCTTCAGGAACGATTGAGGTGATTTCACGAGAAAATGGCTCTGGGACACGGGGTGCCTT CACAGAAATCACAGGGATTCTCAAAAAAGACGGTGATAAAAAAATTGACAACACTGCCAAAACAGCTGT GATTCAAAATAGTACAGAAGGTGTTCTCTCAGCAGTTCAAGGGAATGCTAATGCTATCGGCTACATCTC CTTGGGATCTTTAACGAAATCTGTCAAGGCTTTAGAGATTGATGGTGTCAAGGCTAGTCGAGACACAGT TTTAGATGGTGAATACCCTCTTCAACGTCCCTTCAACATTGTTTGGTCTTCTAATCTTTCCAAGCTAGG TCAAGATTTTATCAGCTTTATCCACTCCAAACAAGGTCAACAAGTGGTCACAGATAATAAATTTATTGA AGCTAAAACCGAAACCACGGAATATACAAGCCAACACTTATCAGGCAAGTTGTCTGTTGTAGGTTCCAC TTCAGTATCTTCTTTAATGGAAAAATTAGCAGAAGCTTATAAAAAAGAAAATCCAGAAGTTACGATTGA TATTACCTCTAATGGGTCTTCAGCAGGTATTACCGCTGTTAAGGAGAAAACCGCTGATATTGGTATGGT TTCTAGGGAATTAACTCCTGAAGAAGGTAAGAGTCTCACCCATGATGCTATTGCTTTAGACGGTATTGC TGTTGTGGTCAATAATGACAATAAGGCAAGCCAAGTCAGTATGGCTGAACTTGCAGACGTTTTTAGTGG CAAATTAACCACCTGGGACAAGATTAAA Table 1 70
SP059 amino acid (SEQ ID NO:98)
KQSASGTIEVISRENGSGTRGAFTEITGILKKDGDKKIDNTAKTAVIQNSTEGVLSAVQGNANAIGYIS LGSLTKSV ALEIDGVKASRDTVLDGEYPLQRPFNIVWSSNLSKLGQDFISFIHSKQGQQWTDNKFIE AKTETTEYTSQHLSGKLSWGSTSVΞSLMEKLAEAYKKENPEVTIDITSNGSSAGITAVKEKTADIGMV SRELTPEEGKSLTHDAIALDGIAVWNNDNKASQVSMAELADVFSGKLTTWDKIK
SP060 nucleotide (SEQ ID NO:99)
ATTCGATGATGCGGATGAAAAGATGACCCGTGATGAAATTGCCTATATGCTGACAAATAGTGAAGAAAC ATTGGATGCTGATGAGATTGAGATGCTACAAGGTGTCTTTTCGCTCGATGAACTGATGGCACGAGAGGT TATGGTTCCTCGAACGGATGCCTTTATGGTGGATATTCAGGATGATAGTCAAGCCATTATCCAAAGTAT TTTAAAACAAAATTATTCTCGTATCCCGGTTTATGATGGGGATAAGGACAATGTAATTGGAATCATTCA CACCAAGAGTCTCCTTAAGGCAGGCTTTGTGGACGGTTTTGACAATATTGTTTGGAAGAGAATTTTACA AGATCCACTTTTTGTACCTGAAACTATTTTTGTGGATGACTTGCTAAAAGAACTGCGAAATACCCAAAG ACAAATG
SP060 amino acid (SEQ ID NO:100)
FDDADEK TRDEIAYMLTNSEETLDADEIEMLQGVFSLDELMAREVMVPRTDAFMVDIQDDSQAIIQSI LKQNYSRIPVYDGDKDNVIGIIHTKSLLKAGFVDGFDNIVWKRILQDPLFVPETIFVDDLLKELR TQR QM
SP062 nucleotide (SEQ ID NO: 101)
GGAGAGTCGATCAAAAGTAGATGAAGCTGTGTCTAAGTTTGAAAAGGACTCATCTTCTTCGTCAAGTTC AGACTCTTCCACTAAACCGGAAGCTTCAGATACAGCGAAGCCAAACAAGCCGACAGAACCAGGAGAAAA GGTAGCAGAAGCTAAGAAGAAGGTTGAAGAAGCTGAGAAAAAAGCCAAGGATCAAAAAGAAGAAGATCG TCGTAACTACCCAACCATTACTTACAAAACGCTTGAACTTGAAATTGCTGAGTCCGATGTGGAAGTTAA AAAAGCGGAGCTTGAACTAGTAAAAGTGAAAGCTAACGAACCTCGAGACGAGCAA
SP062 amino acid (SEQ ID NO:102)
ESRSKVDEAVSKFEKDSSSΞSΞΞDSSTKPEASDTAKPNKPTEPGEKVAEAKKKVEEAEKKAKDQKEEDR RNYPTITYKTLELEIAESDVEVKKAELELVKVKANEPRDEQ
SP063 nucleotide (SEQ ID NO: 103)
ATGGACAACAGGAAACTGGGACGAGGTTATATCTGGTAAGATTGACAAGTACAAAGATCCAGATATTCC AACAGTTGAATCACAAGAAGTTACGTCAGACTCTAGTGATAAAGAAATAACGGTAAGGTATGACCGTTT ATCAACACCAGAAAAACCAATCCCACAACCAAATCCAGAGCATCCAAGTGTTCCGACACCAAACCCAGA ACTACCAAATCAAGAGACTCCAACACCAGATAAACCAACTCCAGAACCAGGTACTCCAAAAACTGAAAC TCCAGTGAATCCAGACCCAGAAGTTCCGACTTATGAGACAGGTAAGAGAGAGGAATTGCCAAACACAGG TACAGAAGCTAAT
SP063 amino acid (SEQ ID NO: 104)
WTTGN DEVIΞGKIDKYKDPDIPTVESQEVTSDSΞDKEITVRYDRLSTPEKPIPQPNPEHPSVPTPNPE LPNQETPTPDKPTPEPGTPKTΞTP PDPEVPTYETGKREELPNTGTEAN
SP064 nucleotide (SEQ ID NO:105)
CGATGGGCTCAATCCAACCCCAGGTCAAGTCTTACCTGAAGAGACATCGGGAACGAAAGAGGGTGACTT ATCAGAAAAACCAGGAGACACCGTTCTCACTCAAGCGAAACCTGAGGGCGTTACTGGAAATACGAATTC ACTTCCGACACCTACAGAAAGAACTGAAGTGAGCGAGGAAACAAGCCCTTCTAGTCTGGATACACTTTT TGAAAAAGATGAAGAAGCTCAAAAAAATCCAGAGCTAACAGATGTCTTAAAAGAAACTGTAGATACAGC TGATGTGGATGGGACACAAGCAAGTCCAGCAGAAACTACTCCTGAACAAGTAAAAGGTGGAGTGAAAGA AAATACAAAAGACAGCATCGATGTTCCTGCTGCTTATCTTGAAAAAGCTGAAGGGAAAGGTCCTTTCAC TGCCGGTGTAAACCAAGTAATTCCTTATGAACTATTCGCTGGTGATGGTATGTTAACTCGTCTATTACT AAAAGCTTCGGATAATGCTCCTTGGTCTGACAATGGTACTGCTAAAAATCCTGCTTTACCTCCTCTTGA AGGATTAACAAAAGGGAAATACTTCTATGAAGTAGACTTAAATGGCAATACTGTTGGTAAACAAGGTCA AGCTTTAATTGATCAACTTCGCGCTAATGGTACTCAAACTTATAAAGCTACTGTTAAAGTTTACGGAAA TAAAGACGGTAAAGCTGACTTGACTAATCTAGTTGCTACTAAAAATGTAGACATCAACATCAATGGATT AGTTGCTAAAGAAACAGTTCAAAAAGCCGTTGCAGACAACGTTAAAGACAGTATCGATGTTCCAGCAGC CTACCTAGAAAAAGCCAAGGGTGAAGGTCCATTCACAGCAGGTGTCAACCATGTGATTCCATACGAACT CTTCGCAGGTGATGGCATGTTGACTCGTCTCTTGCTCAAGGCATCTGACAAGGCACCATGGTCAGATAA Table 1 71
CGGCGACGCTAAAAACCCAGCCCTATCTCCACTAGGCGAAAACGTGAAGACCAAAGGTCAATACTTCTA TCAANTAGCCTTGGACGGAAATGTAGCTGGCAAAGAAAAACAAGCGCTCATTGACCAGTTCCGAGCAAA NGGTACTCAAACTTACAGCGCTACAGTCAATGTCTATGGTAACAAAGACGGTAAACCAGACTTGGACAA CATCGTAGCAACTAAAAAAGTCACTATTAACATAAACGGTTTAATTTCTAAAGAAACAGTTCAAAAAGC CGTTGCAGACAACGTTAANGACAGTATCGATGTTCCAGCAGCCTACCTAGAAAAAGCCAAGGGTGAAGG TCCATTCACAGCAGGTGTCAACCATGTGATTCCATACGAACTCTTCGCAGGTGATGGTATGTTGACTCG TCTCTTGCTCAAGGCATCTGACAAGGCACCATGGTCAGATAACGGNGACGCTAAAAACCCAGCNCTATC TCCACTAGGTGAAAACGTGAAGACCAAAGGTCAATACTTCTATCAANTAGCCTTGGACGGAAATGTAGC TGGCAAAGAAAAACAAGCGCTCATTGACCAGTTCCGAGCAAACGGTACTCAAACTTACAGCGCTACAGT CAATGTCTATGGTAACAAAGACGGTAAACCAGACTTGGACAACATCGTAGCAACTAAAAAAGTCACTAT TAAGATAAATGTTAAAGAAACATCAGACACAGCAAATGGTTCATTATCACCTTCTAACTCTGGTTCTGG CGTGACTCCGATGAATCACAATCATGCTACAGGTACTACAGATAGCATGCCTGCTGACACCATGACAAG TTCTACCAACACGATGGCAGGTGAAAACATGGCTGCTTCTGCTAACAAGATGTCTGATACGATGATGTC AGAGGATAAAGCTATG
SP064 amino acid (SEQ ID NO:106)
DGLNPTPGQVLPEETSGTKEGDLΞΞKPGDTVLTQAKPEGVTGNTNSLPTPTERTEVSEETSPSSLDTLF EKDEEAQKNPELTDVLKETVDTADVDGTQASPAETTPEQVKGGVKENTKDSIDVPAAYLEKAEGKGPFT AGVNQVIPYΞLFAGDGMLTRLLLKASDNAP ΞDNGTAKNPALPP EGLTKGKYFYΞVDLNGNTVGKQGQ ALIDQLRANGTQTYKATVKVYGNKDGKADLTNLVATK1WDININGLVAKETVQKAVADNVKDSIDVPAA YLEKAKGEGPFTAGWHVIPYELFAGDGM TRLLLKASDKAPWSDNGDAKΗPALSPLGENVKTKGQYFY QXALDGNVAGKEKQALIDQFRAXGTQTYSATVNVYGNKDGKPDLDNIVATKKVTININGLISKETVQKA VADNVXDSIDVPAAYLEKAKGEGPFTAGVNHVIPYELFAGDGMLTRLLLKASDKAP SDNGDAKNPALS PLGENVKTKGQYFYQXALDGNVAGKΞKQA IDQFRANGTQTYSATVNVYGNKDGKPDLDNIVATKKVTI KIIWKETSDTANGSLSPSNSGSGVTPMNHNHATGTTDSMPADTMTSSTNT AGENMAASA KMSDTMMS EDKAM
SP065 nucleotide (SEQ ID NO: 107)
TTCCAATCAAAAACAGGCAGATGGTAAACTCAATATCGTGACAACCTTTTACCCTGTCTATGArTTTAC CAAGCAAGTCGCAGGAGATACGGCTAATGTAGAACTCCTAATCGGTGCTGGGACAGAACCTCATGAATA CGAACCATCTGCCAAGGCAGTTGCCAAAATCCAAGATGCAGATACCTTCGTTTATGAAAATGAAAACAT GGAAACATGGGTACCTAAATTGCTAGATACCTTGGATAAGAAAAAAGTGAAAACCATCAAGGCGACAGG CGATATGTTGCTCTTGCCAGGTGGCGAGGAAGAAGAGGGAGACCATGACCATGGAGAAGAAGGTCATCA CCATGAGTTTGACCCCCATGTTTGGTTATCACCAGTTCGTGCCATtAAACTAGTAGAGCACCATCCGCG ACACTTGTCAGCAGATTATCCTGATAAAAAAGAGACCTTTGAGAAGAATGCAGCTGCCTATATCGAAAA ATTGCAAGCCTTGGATAAGGCTTACGCAGAAGGTTTGTCTCAAGCAAAACAAAAGAGCTTTGTGACTCA ACACGCAgCCTTTAACTaTCTTGCCTTGGACTATGGGACTC
SP065 amino acid (SEQ ID NO:108)
SNQKQADGKLNIVTTFYPVYEFTKQVAGDTANVELLIGAGTEPHEYEPSAKAVAKIQDADTFVYENENM ET VPKLLDTLDKKKVKTIKATGDMLLLPGGEEEEGDHDHGEEGHHHEFDPHVWLSPVRAIKLVEHHPR HLSADYPDKKETFEKNAAAYIEKLQALDKAYAEGLSQAKQKSFVTQHAAFNYLA DYGT
SP067 nucleotide (SEQ ID NO: 109)
TATCACAGGATCGAACGGTAAGACAACCACAACGACTATGATTGGGGAAGTTTTGACTGCTGCTGGCCA ACATGGTCTTTTATCAGGGAATATCGGCTATCCAGCTAGTCAGGTTGCTCAAATAGCATCAGATAAGGA CACGCTTGTTATGGAACTTTCTTCTTTCCAACTCATGGGTGTTCAAGAATTCCATCCAGAGATTGCGGT TATTACCAACCTCATGCCAACTCATATCGACTACCATGGGTCATTTTCGGAATATGTAGCAGCCAAGTG GAATATCCAGAACAAGATGACAGCAGCTGATTTCCTTGTCTTGAACTTTAATCAAGACTTGGCAAAAGA CTTGACTTCCAAGACAGAAGCCACTGTTGTACCATTTTCAACACTTGAAAAGGTTGATGGAGCTTATCT GGAAGATGGTCAACTCTACTTCCGTGGTGAAGTAGTCATGGCAGCGAATGAAATCGGTGTTCCAGGTAG CCACAATGTGGAAAATGCCCTTGCGACTATTGCTGTAGCCAAGCTTCGTGATGTGGACAATCAAACCAT CAAGGAAACTCTTTCAGCCTTCGGTGGTGTCAAACACCGTCTCCAGTTTGTGGATGACATCAAGGGTGT TAAATTCTATAACGACAGTAAATCAACTAATATCTTGGCTACTCAAAAAGCCTTGTCAGGATTTGACAA CAGCAAGGTCGTCTTGATTGCAGGTGGTTTGGACCGTGGCAATGAGTTTGACGAATTGGTGCCAGACAT TACTGGACTCAAGAAGATGGTCATCCTGGGTCAATCTGCAGAACGTGTCAAACGGGCAGCAGACAAGGC TGGTGTCGCTTATGTGGAGGCGACAGATATTGCAGATGCGACCCGCAAGGCCTATGAGCTTGCGACTCA Table 1 72
AGGAGATGTGGTTCTTCTTAGTCCTGCCAATGCTAGCTGGGATATGTATGCTAACTTTGAAGTACGTGG CGACCTCTTTATCGACACAGTAGCGGAGTTAAAAGAA
SP067 amino acid (SEQ ID NO: 110)
GITGSNGKTTTTTMIGEVLTAAGQHGLLSGNIGYPAΞQVAQIASDKDT VMELSSFQL GVQEFHPEIA VITNLMPTHIDYHGSFSEYVAAKWNIQNKMTAADFLVLNFNQDLAKD TSKTFJ^TVVPFSTLEKVDGAY LEDGQ YFRGEVV AANEIGVPGSHNVENALATIAVAKLRDVDNQTIKETLSAFGGVKHRLQFVDDIKG VKFYNDSKSTNILATQKALSGFDNSKWLIAGGLDRGNEFDELVPDITGLKKMVI GQSAERVKRAADK AGVAYVEATDIADATRKAYELATQGDW LSPANAS DMYA FEVRGDLFIDTVAE KE
SP068 nucleotide (SEQ ID NO:lll)
AAGTTCATCGAAGATGGTTGGGAAGTCCACTATATCGGGGACAAGTGTGGTATCGAACACCAAGAAATC CTTAAGTCAGGTTTGGATGTCACCTTCCATTCTATTGCGACTGGAAAATTGCGTCGCTATTTCTCTTGG CAAAATATGCTGGACGTCTTCAAAGTTGGTTGGGGAATTGTCCAATCGCTCTTTATCATGTTGCGACTG CGTCCACAGACCCTTTTTTCAAAGGGGGGCTTTGTCTCAGTACCGCCTGTTATCGCTGCGCGTGTGTCA GGAGTGCCTGTCTTTATTCACGAATCTGACCTGTCTATGGGCTTGGCCAATAAAATCGCCTATAAATTT GCGACTAAGATGTATTCAACCTTTGAACAAGCTTCGAGTTTGGCTAAGGTTGAGCATGTGGGAGCGG
SP068 amino acid (SEQ ID NO:112)
SSSK VGKSTISGTSWSNTKKS SQVWMSPSILLRLENCVAISLGKIC TSSKLVGE SNRSLSCCDC VHRPFFQRGALSQYRLLSLRVCQECLSLFTNLTCL AWPIKSPINLRLRCIQPLNKLRV LRLSMWER
SP069 nucleotide (SEQ ID NO:113)
ATCGCTAGCTAGTGAAATGCAAGAAAGTACACGTAAATTCAAGGTTACTGCTGACCTAACAGATGCCGG TGTTGGAACGATTGAAGTTCCTTTGAGCATTGAAGATTTACCCAATGGGCTGACCGCTGTGGCGACTCC GCAAAAAATTACAGTCAAGATTGGTAAGAAGGCTCAGAAGGATAAGGTAAAGATTGTACCAGAGATTGA CCCTAGTCAAATTGATAGTCGGGTACAAATTGAAAATGTCATGGTGTCAGATAAAGAAGTGTCTATTAC GAGTGACCAAGAGACATTGGATAGAATTGATAAGATTATCGCTGTTTTGCCAACTAGCGAACGTATAAC AGGTAATTACAGTGGTTCAGTACCTTTGCAGGCAATCGACCGCAATGGTGTTGTCTTACCGGCAGTTAT CACTCCGTTTGATACAATAATGAAGGTGACTACAAAACCAGTAGCACCAAGTTCAAGCACATCAAATTC AAGTACAAGCAGTTCATCGGAGACATCTTCGTCAACGAAAGCAACTAGTTCAAAAACGAAT
SP069 amino acid (SEQ ID NO:114)
SLASEMQESTRKFKVTADLTDAGVGTIEVPLSIEDLPNGLTAVATPQKITVKIGKKAQKDKVKIVPEID PΞQIDSRVQIΞNλ VSDKEVSITSDQETLDRIDKIIAVLPTSERITGNYΞGSVPLQAIDRNGWLPAVI TPFDTIMKVTTKPVAPSSSTSNΞSTSΞSSETSSSTKATSSKTN
SP070 nucleotide (SEQ ID NO: 115)
GCACCAGATGGGGCACAAGGTTCAGGGATCAGATGTTGAAAAGTACTACTTTACCCAACGCGGTCTTGA GCAGGCAGGAATTACCATTCTTCCTTTTGATGAAAAAAATCTAGACGGTGATATGGAAATTATCGCTGG AAATGCCTTTCGTCCAGATAACAACGTCGAAATTGCCTATGCGGACCAAAATGGTATCAGCTACAAACG TTACCATGAGTTTCTAGGTAGCTTTATGCGTGACTTTGTTAGCATGGGAGTAGCAGGAGCACATGGAAA AACTTCAACGACAGGTATGTTGTCTCATGTCTTGTCTCACATTACAGATACCAGCTTCTTGATTGGAGA TGGGACAGGTCGTGGTTCGGCCAATGCCAAATATTTTGTCTTTGAATCTGACGAATATGAGCGTCACTT CATGCCTTACCACCCAGAATACTCTATTATCACCAACATTGACTTTGACCATCCAGATTATTTCACAAG TCTCGAGGATGTTTTTAATGCCTTTAACGACTATGCCAAACAAATCACCAAGGGTCTTTTTGTCTATGG TGAAGATGCTGAATTGCGTAAGATTACGTCTGATGCACCAATTTATTATTATGGTTTTGAAGCTGAAGG CAATGACTTTGTAGCTAGTGATCTTCTTCGTTCAATAACTGGTTCAACCTTCACCGTTCATTTCCGTGG ACAAAACTTGGGGCAATTCCACATTCCAACCTTTGGTCGTCACAATATCATGAATGCGACAGCCGTTAT TGGTCTTCTTTACACAGCAGGATTTGATTTGAACTTGGTGCGTGAGCACTTGAAAACATTTGCCGGTGT TAAACGTCGTTTCACTGAGAAAATTGTCAATGATACAGTGATTATCGATGACTTTGCCCACCATCCAAC AGAAATTATTGCGACCTTGGATGCGGCTCGTCAGAAATACCCAAGCAAGGAAATTGTAGCAGTCTTTCA ACCGCATACCTTTACAAGAACCATTGCCTTGTTGGACGACTTTGCCCATGCTTTAAACCAAGCAGATGC TGTTTATCTAGCGCAAATTTATGGCTCGGCTCGTGAAGTAGATCATGGTGACGTTAAGGTAGAAGACCT AGCCAACAAAATCAACAAAAAACACCAAGTGATTACTGTTGAAAATGTTTCTCCACTCCTAGACCATGA CAATGCTGTTTACGTCTTTATGGGAGCAGGAGACATCCAAACCTATGAATACTCATTTGAGCGTCTCTT GTCTAACTTGACAAGCAATGTTCAA Table 1 73
SP070 amino acid (SEQ ID NO:116)
HQMGHKVQGSDVEKYYFTQRGLEQAGITILPFDEKNLDGDMEIIAGNAFRPDNNVEIAYADQNGISYKR YHEFLGSFMRDFVSMGVAGAHGKTSTTGMLSHVLSHITDTSFLIGDGTGRGΞANAKYFVFESDEYERHF MPYHPEYSIITNIDFDHPDYFTSLΞDVFNAFNDYAKQITKGLFVYGEDAELRKITSDAPIYYYGFEAEG NDFVASDLLRSITGSTFTVHFRGQNLGQFHIPTFGRHNIMNATAVIGLLYTAGFDLNLVREHLKTFAGV KRRFTEKIVNDTVIIDDFAHHPTEIIATLDAARQKYPSKEIVAVFQPHTFTRTIAL DDFAHALNQADA VΎLAQIYGSAREVDHGDVKVEDLANKINKKHQVITVE.W'SPLLDHDNAVΎVFMGAGDIQTYEYSFERLL SN TSNVQ
SP071 nucleotide (SEQ ID NO:117)
TTTTAACCCAACTGTTGGTACTTTCCTTTTTACTGCAGGATTGAGCTTGTTAGTTTTATTGGTTTCTAA AAGGGAAAATGGAAAGAAACGACTTGTTCATTTTCTGCTGTTGACTAGCATGGGAGTTCAATTGTTGCC GGCCAGTGCTTTTGGGTTGACCAGCCAGATTTTATCTGCCTATAATAGTCAGCTTTCTATCGGAGTCGG GGAACATTTACCAGAGCCTCTGAAAATCGAAGGTTATCAATATATTGGTTATATCAAAACTAAGAAACA GGATAATACAGAGCTTTCAAGGACAGTTGATGGGAAATACTCTGCTCAAAGAGATAGTCAACCAAACTC TACAAAAACATCAGATGTAGTTCATTCAGCTGATTTAGAATGGAACCAAGGACAGGGGAAGGTTAGTTT ACAAGGTGAAGCATCAGGGGATGATGGACTTTCAGAAAAATCTTCTATAGCAGCAGACAATCTATCT C TAATGATTCATTCGCAAGTCAAGTTGAGCAGAATCCGGATCACAAAGGAGAATCTGTAGTTCGACCAAC AGTGCCAGAACAAGGAAATCCTGTGTCTGCTACAACGGTGCAGAGTGCGGAAGAGGAAGTATTGGCGAC GACAAATGATCGACCAGAGTATAAACT CCATTGGAAACCAAAGGCACGCAAGAACCCGGTCATGAGGG TGAAGCCGCAGTCCGTGAAGACTTACCAGTCTACACTAAGCCACTAGAAACCAAAGGTACACAAGGACC CGGACATGAAGGTGAAGCTGCAGTTCGCGAGGAAGAACCAGCTTACACAGAACCGTTAGCAACGAAAGG CACGCAAGAGCCAGGTCATGAGGGCAAAGCTACAGTCCGCGAAGAGACTCTAGAGTACACGGAACCGGT AGCGACAAAAGGCACACAAGAACCCGAACATGAGGGCGAACGGSCAGTAGAAGAAGAACTTCCGGCTTT AGAGGTCACTACACGAAATAGAACGGAAATCCAGAATATTCCTTATACAACAGAAGAAATTCAGGATCC AACACTTCTGAAAAATCGTCGTAAGATTGAACGACAAGGGCAAGCAGGGACACGTACAATTCAATATGA AGACTACATCGTAAATGGTAATGTCGTAGAAACTAAAGAAGTGTCACGAACTGAAGTAGCTCCGGTCAA CGAAGTCGTTAAAGTAGGAACACTTGTGAAAGTTAAACCTACAGTAGAAATTACAAACTTAACAAAAGT TGAGAACAAAAAATCTATAACTGTAAGTTATAACTTAATAGACACTACCTCAGCATATGTTTCTGCAAA AACGCAAGTTTTCCATGGAGACAAGCTAGTTAAAGAGGTGGATATAGAAAATCCTGCCAAAGAGCAAGT ATATCAGGTTTAGATTACTACACACCGTATACAGTTAAAACACACCTAACTTATAATTTGGGTGAAAA TAATGAGGAAAATACTGAAACATCAACTCAAGATTTCCAATTAGAGTATAAGAAAATAGAGATTAAAGA ATTGATTCAGTAGAATTATACGGTAAAGAAAATGATCGTTATCGTAGATATTTAAGTCTAAGTGAAGC GCCGACTGATACGGCTAAATACTTTGTAAAAGTGAAATCAGATCGCTTCAAAGAAATGTACCTACCTGT AAAATCTATTACAGAAAATACGGATGGAACGTATAAAGTGACGGTAGCCGTTGATCAACTTGTCGAAGA AGGTACAGACGGTTACAAAGATGATTACACATTTACTGTAGCTAAATCTAAAGCAGAGCAACCAGGAGT TACACATCCTTTAAACAGCTGGTAACAGCCATGCAAAGCAATCTGTCTGGTGTCTATACATTGGCTTC AGATATGACCGCAGATGAGGTGAGCTTAGGCGATAAGCAGACAAGTTATCTCACAGGTGCATTTACAGG GAGCTTGATCGGTTCTGATGGAACAAAATCGTATGCCATTTATGATTTGAAGAAACCATTATTTGATAC ATTAAATGGTGCTACAGTTAGAGATTTGGATATTAAAACTGTTTCTGCTGATAGTAAAGAAAATGTCGC GCGCTGGCGAAGGCAGCGAATAGCGCGAATATTAATAATGTTGCAGTAGAAGGAAAAATCTCAGGTGC GAAATCTGTTGCGGGATTAGTAGCGAGCGCAACAAATACAGTGATAGAAAACAGCTCGTTTACAGGGAA ACTTATCGCAAATCACCAGGACAGTAATAAAAATGATACTGGAGGAATAGTAGGTAATATAACAGGAAA TAGTTCGAGAGTTAATAAAGTTAGGGTAGATGCCTTAATCTCTACTAATGCACGCAATAATAACCAAAC AGCTGGAGGGATAGTAGGTAGATTAGAAAATGGTGCATTGATATCTAATTCGGTTGCTACTGGAGAAAT ACGAAATGGTCAAGGATATTCTAGAGTCGGAGGAATAGTAGGATCTACGTGGCAAAACGGTCGAGTAAA TAATGTTGTGAGTAACGTAGATGTTGGAGATGGTTATGTTATCACCGGTGATCAATACGCAGCAGCAGA TGTGAAAAATGCAAGTACATCAGTTGATAATAGAAAAGCAGACAGATTCGCTACAAAATTATCAAAAGA CCAAATAGACGCGAAAGTTGCTGATTATGGAATCACAGTAACTCTTGATGATACTGGGCAAGATTTAAA ACGTAATCTAAGAGAAGTTGATTATACAAGACTAAATAAAGCAGAAGCTGAAAGAAAAGTAGCTTATAG CAACATAGAAAAACTGATGCCATTCTACAATAAAGACCTAGTAGTTCACTATGGTAACAAAGTAGCGAC AACAGATAAACTTTACACTACAGAATTGTTAGATGTTGTGCCGATGAAAGATGATGAAGTAGTAACGGA TATTAATAATAAGAAAAATTCAATAAATAAAGTTATGTTACATTTCAAAGATAATACAGTAGAATACCT AGATGTAACATTCAAAGAAAACTTCATAAACAGTCAAGTAATCGAATACAATGTTACAGGAAAAGAATA TATATTCACACCAGAAGCATTTGTTTCAGACTATACAGCGATAACGAATAACGTACTAAGCGACTTGCA AAATGTAACACTTAAC
SP071 amino acid (SEQ ID NO:118) Table 1 74
FNPTVGTF FTAGLSL VLLVSKRENGKKRLVHFLLLTSMGVQLLPAΞAFGLTSQILSAYNSQLSIGVG EH PEPLKIEGYQYIGYIKTKKQDNTELSRTVΌGKYSAQRDSQPNSTKTSDWHSADLEWNQGQGKVSL QGEAΞGDDGLSEKSSIAADNLSSNDSFASQVEQNPDHKGESWRPTVPEQGNPVSATTVQSAEEEVLAT TNDRPEYK PLETKGTQEPGHEGEAAVREDLPVYTKPLETKGTQGPGHEGEAAVREEEPAYTEPLATKG TQEPGHEGKATVREETLEYTEPVATKGTQEPEHEGERXVEEELPALEVTTR RTEIQNIPYTTEEIQDP TLLKNRRKIERQGQAGTRTIQYEDYIVNGLWVETKEVSRTEVAPVNEΛ/VKVGTLVKVKPTVEITNLTKV ENKKSITVSYNLIDTTSAYVSAKTQVFHGDKLVKEVDIENPAKEQVISGLDYYTPYTVKTH TYNLGEN NEENTETSTQDFQLEYKKIEIKDIDSVELYGKENDRYRRYLSLΞEAPTDTAKYFVKVKSDRFKΞMYLPV KΞITENTDGTYKVTVAVDQLVEEGTDGYKDDYTFTVAKSKAEQPGVYTSFKQLVTAMQSNLSGVYTLAS DMTADEVSLGDKQTSYLTGAFTGΞLIGSDGTKSYAIYDLKKPLFDTLNGATVRDLDIKTVSADSKENVA ALAKAANSANINNVAVEGKISGAKSVAGLVASATNTVIENΞSFTGKLIANHQDSNKNDTGGIVGNITGN SSRVNKVRVDALISTNARNNNQTAGGIVGRLENGALISNSVATGEIRNGQGYSRVGGIVGST QNGRVN NWSNVDVGDGYVITGDQYAAADVKNASTSVDNRKADRFATKLSKDQIDAKVADYGITVT DDTGQDLK RNLREVDYTRLNKAEAERKVAYSNIEK MPFYNKDLWHYGNKVATTDKLYTTE LDWPMKDDEWTD INNKKNSINKV LHFKDNTVEYLDVTFKENFINSQVIEYNVTGKEYIFTPEAFVSDYTAITNNVLSDLQ
NVTLN
SP072 nucleotide (SEQ ID NO: 119)
TTTTAACCCAACTGTTGGTACTTTCCTTTTTACTGCAGGATTGAGCTTGTTAGTTTTATTGGTTTCTAA AAGGGAAAATGGAAAGAAACGACTTGTTCATTTTCTGCTGTTGACTAGCATGGGAGTTCAATTGTTGCC GGCCAGTGCTTTTGGGTTGACCAGCCAGATTTTATCTGCCTATAATAGTCAGCTTTCTATCGGAGTCGG GGAACATTTACCAGAGCCTCTGAAAATCGAAGGTTATCAATATATTGGTTATATCAAAACTAAGAAACA GGATAATACAGAGCTTTCAAGGACAGTTGATGGGAAATACTCTGCTCAAAGAGATAGTCAACCAAACTC TACAAAAACATCAGATGTAGTTCATTCAGCTGATTTAGAATGGAACCAAGGACAGGGGAAGGTTAGTTT ACAAGGTGAAGCATCAGGGGATGATGGACTTTCAGAAAAATCTTCTATAGCAGCAGACAATCTATCTTC TAATGATTCATTCGCAAGTCAAGTTGAGCAGAATCCGGATCACAAAGGAGAATCTGTAGTTCGACCAAC AGTGCCAGAACAAGGAAATCCTGTGTCTGCTACAACGGTGCAGAGTGCGGAAGAGGAAGTATTGGCGAC GACAAATGATCGACCAGAGTATAAACTTCCATTGGAAACCAAAGGCACGCAAGAACCCGGTCATGAGGG TGAAGCCGCAGTCCGTGAAGACTTACCAGTCTACACTAAGCCACTAGAAACCAAAGGTACACAAGGACC CGGACATGAAGGTGAAGCTGCAGTTCGCGAGGAAGAACCAGCTTACACAGAACCGTTAGCAACGAAAGG CACGCAAGAGCCAGGTCATGAGGGCAAAGCTACAGTCCGCGAAGAGACTCTAGAGTACACGGAACCGGT AGCGACAAAAGGCACACAAGAACCCGAACATGAGGGCGAaCGGsCAGTAGAAGAAGAACTTCCGGCTTT AGAGGTCACTACACGAAATAGAACGGAAATCCAGAATATTCCTTATACAACAGAAGAAATTCAGGATCC AACACTTCTGAAAAATCGTCGTAAGATTGAACGACAAGGGCAAGCAGGGACACGTACAATTCAATATGA AGACTACATCGTAAATGGTAATGTCGTAGAAACTAAAGAAGTGTCACGAACTGAAGTAGCTCCGGTCAA CGAAGTCGTTAAAGTAGGAACACTTGTGAAAGTTAAACCTACAGTAGAAATTACAAACTTAACAAAAGT TGAGAACAAAAAATCTATAACTGTAAGTTATAACTTAATAGACACTACCTCAGCATATGTTTCTGCAAA AACGCAAGTTTTCCATGGAGACAAGCTAGTTAAAGAGGTGGATATAGAAAATCCTGCCAAAGAGCAAGT AATATCAGGTTTAGATTACTACACACCGTATACAGTTAAAACACACCTAACTTATAATTTGGGTGAAAA TAATGAGGAAAATACTGAAACATCAACTCAAGATTTCCAATTAGAGTATAAGAAAATAGAGATTAAAGA TATTGATTCAGTAGAATTATACGGTAAAGAAAATGATCGTTATCGTAGA
SP072 amino acid (SEQ ID NO:120)
FNPTVGTFLFTAGLSLLVLLVSKRENGKKRLVHFLLLTSMGVQLLPASAFGLTSQILSAYNSQLSIGVG EH PEPLKIEGYQYIGYIKTKKQDNTELΞRTVDGKYSAQRDSQPNSTKTSDWHSADLE NQGQGKVSL QGEASGDDGLSEKSSIAADNLSSNDSFASQVEQNPDHKGESWRPTVPEQGNPVSATTVQSAEEEVLAT TNDRPEYKLPLETKGTQEPGHEGEAAVREDLPVYTKPLETKGTQGPGHEGEAAλ/REEEPAYTEPLATKG TQEPGHEGKATVREETLEYTEPVATKGTQEPEHEGERXVEEELPALEVTTRNRTEIQNIPYTTΞEIQDP TL KNRRKIERQGQAGTRTIQYEDYIVNGNWETKEVSRTEVAPλΛIEWKVGTLVKVKPTVEITNLTKV ENKKSITVΞYNLIDTTSAYVSAKTQVFHGDKLV EVDIENPAKEQVISGLDYYTPYTVKTHLTYNLGEN NEENTETSTQDFQLEYKKIEIKDIDSVE YGKENDRYRR
SP073 nucleotide (SEQ ID NO:121)
TCGTAGATATTTAAGTCTAAGTGAAGCGCCGACTGATACGGCTAAATACTTTGTAAAAGTGAAATCAGA TCGCTTCAAAGAAATGTACCTACCTGTAAAATCTATTACAGAAAATACGGATGGAACGTATAAAGTGAC GGTAGCCGTTGATCAACTTGTCGAAGAAGGTACAGACGGTTACAAAGATGATTACACATTTACTGTAGC TAAATCTAAAGCAGAGCAACCAGGAGTTTACACATCCTTTAAACAGCTGGTAACAGCCATGCAAAGCAA TCTGTCTGGTGTCTATACATTGGCTTCAGATATGACCGCAGATGAGGTGAGCTTAGGCGATAAGCAGAC Table 1 75
AAGTTATCTCACAGGTGCATTTACAGGGAGCTTGATCGGTTCTGATGGAACAAAATCGTATGCCATTTA TGATTTGAAGAAACCATTATTTGATACATTAAATGGTGCTACAGTTAGAGATTTGGATATTAAAACTGT TTCTGCTGATAGTAAAGAAAATGTCGCAGCGCTGGCGAAGGCAGCGAATAGCGCGAATATTAATAATGT TGCAGTAGAAGGAAAAATCTCAGGTGCGAAATCTGTTGCGGGATTAGTAGCGAGCGCAACAAATACAGT GATAGAAAACAGCTCGTTTACAGGGAAACTTATCGCAAATCACCAGGACAGTAATAAAAATGATACTGG AGGAATAGTAGGTAATATAACAGGAAATAGTTCGAGAGTTAATAAAGTTAGGGTAGATGCCTTAATCTC TACTAATGCACGCAATAATAACCAAACAGCTGGAGGGATAGTAGGTAGATTAGAAAATGGTGCATTGAT ATCTAATTCGGTTGCTACTGGAGAAATACGAAATGGTCAAGGATATTCTAGAGTCGGAGGAATAGTAGG ATCTACGTGGCAAAACGGTCGAGTAAATAATGTTGTGAGTAACGTAGATGTTGGAGATGGTTATGTTAT CACCGGTGATCAATACGCAGCAGCAGATGTGAAAAATGCAAGTACATCAGTTGATAATAGAAAAGCAGA CAGATTCGCTACAAAATTATCAAAAGACCAAATAGACGCGAAAGTTGCTGATTATGGAATCACAGTAAC TCTTGATGATACTGGGCAAGATTTAAAACGTAATCTAAGAGAAGTTGATTATACAAGACTAAATAAAGC AGAAGCTGAAAGAAAAGTAGCTTATAGCAACATAGAAAAACTGATGCCATTCTACAATAAAGACCTAGT AGTTCACTATGGTAACAAAGTAGCGACAACAGATAAACTTTACACTACAGAATTGTTAGATGTTGTGCC GATGAAAGATGATGAAGTAGTAACGGATATTAATAATAAGAAAAATTCAATAAATAAAGTTATGTTACA TTTCAAAGATAATACAGTAGAATACCTAGATGTAACATTCAAAGAAAACTTCATAAACAGTCAAGTAAT CGAATACAATGTTACAGGAAAAGAATATATATTCACACCAGAAGCATTTGTTTCAGACTATACAGCGAT AACGAATAACGTACTAAGCGACTTGCAAAATGTAACACTTAAC
SP073 amino acid (SEQ ID NO:122)
RRYLSLSEAPTDTAKYFVKVKSDRFKE YLPVKSITENTDGTYKVTVAVDQLVEEGTDGYKDDYTFTVA KSKAEQPGVYTSFKQLVTAMQSNLSGVYTLASDMTADEVSLGDKQTSYLTGAFTGSLIGSDGTKSYAIY DLKKPLFDTLNGATVRDLDIKTVSADSKENVAAliAKAANSANINNVAVEGKISGAKSVAGLVASATNTV IENSSFTGKLIA-fflQDSNKNDTGGIVGNITGNSSRVNKVRVDALISTNARMSTNQTAGGIVGRLENGALI SNSVATGEIRNGQGYSRVGGIVGSTWQNGRVNNλΛ/SNVDVGDGYVITGDQYAAADVKNASTSλπDNRKAD RFATKLSKDQIDAKVADYGITVTLDDTGQDLKRNLREVDYTRLNKAEAERKVAYSNIEK MPFYNKDLV VHYGNKVATTDKLYTTELLDWPMKDDEWTDINNKKNSINKVMLHFKDNTVEYLDVTFKΞNFINSQVI EYNVTGKEYIFTPEAFVSDYTAIT NVLSDLQNVTLN
SP074 nucleotide (SEQ ID NO: 123)
CTTTGGTTTTGAAGGAAGTAAGCGTGGACAATTTGCTGTAGAAGGAATCAATCAACTTCGTGAGCATGT AGACACTCTATTGATTATCTCAAACAACAATTTGCTTGAAATTGTTGATAAGAAAACACCGCTTTTGGA GGCTCTTAGCGAAGCGGATAACGTTCTTCGTCAAGGTGTTCAAGGGATTACCGATTTGATTACCAATCC AGGATTGATTAACCTTGACTTTGCCGATGTGAAAACGGTAATGGCAAACAAAGGGAATGCTCTTATGGG TATTGGTATCGGTAGTGGAGAAGAACGTGTGGTAGAAGCGGCACGTAAGGCAATCTATTCACCACTTCT TGAAACAACTATTGACGGTGCTGAGGATGTTATCGTCAACGTTACTGGTGGTCTTGACTTAACCTTGAT TGAGGCAGAAGAGGCTTCACAAATTGTGAACCAGGCAGCAGGTCAAGGAGTGAACATCTGGCTCGGTAC TTCAATTGATGAAAGTATGCGTGATGAAATTCGTGTAACAGTTGTTGCAACGGGTGTTCGTCAAGACCG CGTAGAAAAGGTTGTGGCTCCACAAGCTAGATCTGCTACTAACTACCGTGAGACAGTGAAACCAGCTCA TTCACATGGCTTTGATCGTCATTTTGATATGGCAGAAACAGTTGAATTGCCAAAACAAAATCCACGTCG TTTGGAACCAACTCAGGCATCTGCTTTTGGTGATTGGGATCTTCGCCGTGAATCGATTGTTCGTACAAC AGATTCAGTCGTTTCTCCAGTCGAGCGCTTTGAAGCCCCAATTTCACAAGATGAAGATGAATTGGATAC ACCTCCATTTTTCAAAAATCGT
SP074 amino acid (SEQ ID NO:124)
FGFEGSKRGQFAVEGINQLREHVDTLLIISNNNL EIVDKKTPL EALSEAD VLRQGVQGITDLITNP GLINLDFADV TVMA KGNALMGIGIGSGEERWEAARKAIYSPLLETTIDGAEDVIVNVTGGLD TLI FAEEASQIVNQAAGQGVNIWLGTSIDESMRDEIRVTVVATGVRQDRVEKVVAPQARSATNYRETVKPAH SHGFDRHFDMAETVELPKQNPRRLEPTQASAFGDWDLRRESIVRTTDSWSPVERFEAPISQDEDELDT PPFFKNR
SP075 nucleotide (SEQ ID NO: 125)
CTACTACCTCTCGAGAGAAAGTGACCTAGAGGTGACCGTTTTTGACCATGAGCAAGGTCAAGCCACCAA GGCCGCAGCAGGAATTATCAGTCCTTGGTTTTCCAAACGCCGTAATAAAGCCTGGTACAAGATGGCGCG CTTGGGGGCTGATTTTTATGTGGATTTATTAGCTGATTTAGAGAAATCAGGACAAGAAATCGACTTTTA CCAGCGTTCGGGAGTCTTTCTCTTGAAAAAGGATGAATCCAATTTGGAAGAACTTTATCAACTGGCCCT CCAGCGCAGAGAAGAATCTCCCTTGATAGGGCAATTAGCCATTCTGAACCAAGCCTCAGCTAATGAATT ATTCCCTGGTTTGCAGGGATTTGACCGCCTGCTCTATGCTTCTGGTGGAGCGAGAGTAGATGGCCAACT Table 1 76
TTTAGTGACTCGTTTGCTGGAAGTCAGTCATGTCAAGCTGGTCAAAGAAAAAGTGACTCTGACACCGTT AGCATCAGGCTACCAGATTGGTGAAGAGGAGTTTGAGCAGGTTATTTTGGCGACGGGAGCTTGGTTGGG GGACATGTTAGAGCCTTTAGGTTATGAAGTGGATGTCCGTCCTCAAAAAGGACAACTACGAGATTATCA GCTTGCCCAAGACATGGAAGATTACCCTGTTGTCATGCCAGAAGGGGAGTGGGATTTGATTCCCTTTGC AGGTGGGAAATTATCCTTAGGCGCTACCCACGAAAATGACATGGGATTTGATTTGACGGTAGATGAAAC CTTGCTCCAACAAATGGAGGAGGCCACCTTGACTCACTATCTGATTTTGGCTGAAGCTACTTCAAAATC TGAGCGTGTTGGAATCCGTGCCTACACCAGTGATTTCTCTCCTTTCTTTGGGCAGGTGCCTGACTTAAC TGGTGTCTATGCAGCCAGTGGACTAGGTTCATCAGGCCTCACAACTGGTCCTATCATTGGTTACCATCT AGCCCAACTGATCCAAGACAAGGAGTTGACCTTGGACCCTCTAAATTACCCAATTGAAAACTATGTCAA ACGAGTAAAAAGCGAA
SP075 amino acid (SEQ ID NO: 126)
YYLSREΞDLEVTVFDHEQGQATKAAAGIISP FSKRR KA YKMARLGADFYVDLLADLEKSGQEIDFY QRSGVF LKKDEΞNLEELYQLALQRREESPLIGQLAILNQASANELFPG QGFDRLLYASGGARVϋGQL LVTRLLEVΞHVKLVKEKVTLTPLASGYQIGEEΞFEQVILATGA LGDMLEPLGYEVDVRPQKGQLRDYQ LAQDMEDYPWMPEGEWDLIPFAGGKLSLGATHEND GFDLTVDETLLQQMEEATLTHYLILAEATSKS ERVGIRAYTSDFSPFFGQVPDLTGVYAASGLGSSG TTGPIIGYHLAQLIQDKELTLDPLNYPIENYVK RVKSE
SP076 nucleotide (SEQ ID NO:127)
TAAGGTCAAAAGTCAGACCGCTAAGAAAGTGCTAGAAAAGATTGGAGCTGACTCGGTTATCTCGCCAGA GTATGAAATGGGGCAGTCTCTAGCACAGACCATTCTTTTCCATAATAGTGTTGATGTCTTTCAGTTGGA TAAAAATGTGTCTATCGTGGAGATGAAAATTCCTCAGTCTTGGGCAGGTCAAAGTCTGAGTAAATTAGA CCTCCGTGGCAAATACAATCTGAATATTTTGGGTTTCCGAGAGCAGGAAAATTCCCCATTGGATGTTGA ATTTGGACCAGATGACCTCTTGAAAGCAGATACCTATATTTTGGCAGTCATCAACAACCAGTATTTGGA TACCCTA
SP076 amino acid (SEQ ID NO:128)
KVKSQTAKKVLEKIGADSVISPEYΞMGQSLAQTILFHNSVOVFQLDKNVSIVEMKIPQSWAGQSLSKLD LRGKYNLNI GFREQENSPLDVEFGPDDLLKADTYILAVINNQYLDTL
SP077 nucleotide (SEQ ID NO:129)
TGACGGGTCTCAGGATCAGACTCAGGAAATCGCTGAGTGTTTAGCTAGCAAGTATCCTAATATCGTTAG AGCCATCTATCAGGAAAATAAATGCCATGGCGGTGCGGTCAATCGTGGCTTGGTAGAGGCTTCTGGGCG CTATTTTAAAGTAGTTGACAGTGATGACTGGGTGGATCCTCGTGCCTACTTGAAAATTCTTGAAACTTG CAGGAACTTGAGAGCAAAGGTCAAGAGGTGGATGTCTTTG
SP077 amino acid (SEQ ID NO: 130)
DGSQDQTQEIAECLASKYPNIVRAIYQENKCHGGAVNRGLVEASGRYFKVVυSDDWVDPRAYLKILETC RNLRAKVKRWMSL
SP078 nucleotide (SEQ ID NO:131)
TAGAGGCTTTGCCAAATGGTGGGAAGGGCACGAGCGTCGAAAAGAGGAACGCTTTGTCAAACAAGAAGA AAAAGCTCGCCAAAAGGCTGAGAAAGAGGCTAGATTAGAACAAGAAGAGACTGAAAAAGCCTTACTCGA TTTGCCTCCTGTTGATATGGAAACGGGTGAAATTCTGACAGAGGAAGCTGTTCAAAATCTTCCACCTAT TCCAGAAGAAAAGTGGGTGGAACCAGAAATCATCCTGCCTCAAGCTGAACTTAAATTCCCTGAACAGGA AGATGACTCAGATGACGAAGATGTTCAGGTCGATTTTTCAGCCAAAGAAGCCCTTGAATACAAACTTCC AAGCTTACAACTCTTTGCACCAGATAAACCAAAAGATCAGTCTAAAGAGAAGAAAATTGTCAGAGAAAA TATCAAAATCTTAGAAGCAACCTTTGCTAGCTTTGGTATTAAGGTAACAGTTGAACGGGCCGAAATTGG GCCATCAGTGACCAAGTATGAAGTCAAGCCGGCTGTTGGTGTAAGGGTCAACCGCATTTCCAATCTATC AGATGACCTCGCTCTAGCCTTGGCTGCCAAAGATGTCCGGATTGAAGCACCAATCCCTGGGAAATCCCT AATCGGAATTGAAGTGCCCAACTCCGATATTGCCACTGTATCTTTCCGAGAACTATGGGAACAATCGCA AACGAAAGCAGAAAATTTCTTGGAAATTCCTTTAGGGAAGGCTGTTAATGGAACCGCAAGAGCTTTTGA CCTTTCTAAAATGCCCCACTTGCTAGTTGCAGGTTCAACGGGTTCAGGGAAGTCAGTAGCAGTTAACGG CATTATTGCTAGCATTCTCATGAAGGCGAGACCAGATCAAGTTAAATTTATGATGGTCGATCCCAAGAT GGTTGAGTTATCTGTTTACAATGATATTCCCCACCTCTTGATTCCAGTCGTGACCAATCCACGCAAAGC CAGCAAGGCTCTGCAAAAGGTTGTGGATGAAATGGAAAACCGTTATGAACTCTTTGCCAAGGTGGGAGT TCGGAATATTGCAGGTTTTAATGCCAAGGTAGAAGAGTTCAATTCCCAGTCTGAGTACAAGCAAATTCC Table 1 77
GCTACCATTCATTGTCGTGATTGTGGATGAGTTGGCTGACCTCATGATGGTGGCCAGCAAGGAAGTGGA AGATGCTATCATCCGTCTTGGGCAGAAGGCGCGTGCTGCAGGTATCCACATGATTCTTGCAACTCAGCG TCCATCTGTTGATGTCATCTCTGGTTTGATTAAGGCCAATGTTCCATCTCGTGTAGCATTTGCGGTTTC ATCAGGAACAGACTCCCGTACGATTTTGGATGAAAATGGAGCAGAAAAACTTCTTGGTCGAGGAGACAT GCTCTTTAAACCGATTGATGAAAATCATCCAGTTCGTCTCCAAGGCTCCTTTATCTCGGATGACGATGT TGAGCGCATTGTGAACTTCATCAAGACTCAGGCAGATGCAGACTACGATGAGAGTTTTGATCCAGGTGA GGTTTCTGAAAATGAAGGAGAATTTTCGGATGGAGATGCTGGTGGTGATCCGCTTTTTGAAGAAGCTAA GTCTTTGGTTATCGAAACACAGAAAGCCAGTGCGTCTATGATTCAGCGTCGTTTATCAGTTGGATTTAA CCGTGCGACCCGTCTCATGGAAGAACTGGAGATAGCAGGTGTCATCGGTCCAGCTGAAGGTACCAAACC TCGAAAAGTGTTACAACAA
SP078 amino acid (SEQ ID NO: 132)
RGFAK EGHERRKEERFVKQEEKARQKAEKEAR EQEETEKA LDLPPVDMETGEI TEEAVQNLPPI PEEK VEPEIILPQAELKFPEQEDDSDDEDVQVDFSAKEALEYKLPSLQLFAPDKPKDQSKEKKIVREN IKI EATFASFGIKVTVERAEIGPSVTKYEVKPAVGVRVNRISNLΞDDLALALAAKDVRIEAPIPGKSL IGIEVPNSDIATVSFRELWEQSQTKAENF EIPLGKAVNGTARAFDLSKMPHLLVAGSTGSGKSVAVNG IIASILMKARPDQVKFMMVDPKMVELSVYNDIPHL IPWTNPRKASKALQKWDEMENRYE FAKVGV RNIAGFNAKVEEFNSQSEYKQIPLPFIVVIVDELADL MVAΞKEVEDAIIRLGQKARAAGIHMILATQR PSVDVISGLIKANVPSRVAFAVSΞGTDSRTILDENGAEKLLGRGDMLFKPIDENHPVRLQGSFISDDDV ERIVNFIKTQADADYDESFDPGEVSENEGEFSDGDAGGDPLFEEAKSLVIETQKASASMIQRRLSVGFN RATRL EELEIAGVIGPAEGTKPRKVLQQ
SP079 nucleotide (SEQ ID NO:133)
TCAAAAAGAGAAGGAAAACTTGGTTATTGCTGGGAAAATAGGTCCAGAACCAGAAATTTTGGCCAATAT GTATAAGTTGCTGAT^GAAGAAAATACCAGCATGACTGCGACTGTTAAACCGAATTTTGGGAAGACAAG CTTCCTTTATGAAGCTCTGAAAAAAGGCGATATTGACATCTATCCTGAATTTACTGGTACGGTGACTGA AAGTTTGCTTCAACCATCACCCAAGGTGAGTCATGAACCAGAACAGGTTTATCAGGTGGCGCGTGATGG CATTGCTAAGCAGGATCATCTAGCCTATCTCAAACCCATGTCTTATCAAAACACCTATGCTGTAGCTGT TCCGAAAAAGATTGCTCAAGAATATGGCTTGAAGACCATTTCAGACTTGAAAAAAGTGGAAGGGCAGTT GAAGGCAGGTTTTACACTCGAGTTTAACGACCGTGAAGATGGAAATAAGGGCTTGCAATCAATGTATGG TCTCAATCTCAATGTAGCGACCATTGAGCCAGCCCTTCGCTATCAGGCTATTCAGTCAGGGGATATTCA AATCACGGATGCCTATTCGACTGATGCGGAATTGGAGCGTTATGATTTACAGGTCTTGGAAGATGACAA GCAACTCTTCCCACCTTATCAAGGGGCTCCACTCATGAAAGAAGCTCTTCTCAAGAAACACCCAGAGTT GGAAAGAGTTCTTAATACATTGGCTGGTAAGATTACAGAAAGCCAGATGAGCCAGCTCAACTACCAAGT CGGTGTTGAAGGCAAGTCAGCAAAGCAAGTAGCCAAGGAGTTTCTCCAAGAACAAGGTTTGTTGAAGAA A
SP079 amino acid (SEQ ID NO:134)
QKEKENLVIAGKIGPEPEILANMYKLLIEENTSMTATVKPNFGKTSFLYEALKKGDIDIYPEFTGTVTE SLLQPSPKVSHEPEQVYQVARDGIAKQDH AYLKPMSYQNTYAVAVPKKIAQEYGLKTISDLKKVEGQL KAGFTLEFNDREDGNKGLQSMYGLNLNVATIEPALRYQAIQSGDIQITDAYSTDAELERYDLQVLEDDK QLFPPYQGAPLMKEA LKKHPELERVLNTLAGKITESQMSQLNYQVGVEGKSAKQVAKEFLQEQG LKK
SP080 nucleotide (SEQ ID NO:135)
ACGTTCTATTGAGGACCACTTTGATTCAAACTTCGAATTGGAATATAACCTCAAAGAAAAAGGGAAAAC AGATCTTTTGAAGCTAGTTGATAAAACAACTGACATGCGTCTGCATTTTATCCGCCAAACTCATCCACG CGGTCTCGGAGATGCTGTTTTGCAAGCCAAGGCTTTCGTCGGAAATGAACCTTTTGTCGTTATGCTTGG TGATGACTTGATGGATATCACAGACGAAAAGGCTGTTCCACTTACCAAACAACTCATGGATGACTACGA GCGTACCCACGCGTCTACTATCGCTGTCATGCCAGTCCCTCATGACGAAGTATCTGCTTACGGGGTTAT TGCTCCGCAAGGCGAAGGAAAAGATGGTCTTTACAGTGTTGAAACCTTTGTTGAAAAACCAGCTCCAGA GGACGCTCCTAGCGACCTTGCTATTATCGGACGCTACCTCCTCACGCCTGAAATTTTTGAGATTCTCGA AAAGCAAGCTCCAGGTGCAGGAAATGAAATTCAGCTGACAGATGCAATCGACACCCTCAATAAAACACA ACGTGTATTTGCTCGTGAGTTCAAAGGGGCTCGTTACGATGTCGGAGACAAGTTTGGCTTCATGAAAAC ATCCATCGACTACGCCCTCAAACACCCACAAGTCAAAGATGATTTGAAGAATTACCTCATCCAACTTGG AAAAGAATTGACTGAGAAGGAA Table 1 78
SP080 amino acid (SEQ ID NO: 136)
RSIEDHFDSNFELEYNLKEKGKTDLLKLVDKTTDMR HFIRQTHPRGLGDAVLQAKAFVGNEPFWMLG DDLMDITDEKAVPLTKQLMDDYERTHASTIAVMPVPHDEVSAYGVIAPQGEGKDGLYSVETFVEKPAPE DAPSDLAIIGRYLLTPEIFEILEKQAPGAGNEIQLTDAIDTLNKTQRVFAREFKGARYDVGDKFGF KT SIDYALKHPQVKDDLKNYLIQLGKELTEKE
SP081 nucleotide (SEQ ID NO:137)
CGCTCAAAATACCAGAGGTGTTCAGCTAATCGAGCACGTTTCTCCTCAAATGTTGAAAGCCCAATTGGA GAGTGTCTTTTCTGATATTCCACCTCAGGCTGTAAAAACTGGAATGTTGGCTACTACTGAAATCATGGA AATCATCCAACCCTATCTTAAAAAACTGGATTGTCCCTATGTCCTTGATCCTGTTATGGTTGCTACAAG TGGAGATGCCTTGATTGACTCAAATGCTAGAGACTATCTCAAAACAAACTTACTACCTCTAGCAACTAT TATTACGCCAAATCTTCCTGAAGCAGAAGAGATTGTTGGTTTTTCAATCCATGACCCCGAAGACATGCA GCGTGCTGGTCGCCTGATTTTAAAAGAATTTGGTCCTCAGTCTGTGGTTATCAAAGGCGGACATCTCAA AGGTGGTGCTAAAGATTTCCTCTTTACCAAGAATGAACAATTTGTCTGGGAAAGCCCACGAATTCAAAC CTGTCACACCCATGGTACT
SP081 amino acid (SEQ ID NO:138)
AQNTRGVQLIEHVSPQMLKAQLESVFSDIPPQAVKTGMLATTEIMEIIQPYLKKLDCPYV DPVMVATS GDALIDSNARDYLKTNLLPLATIITPNLPEAEEIVGFSIHDPEDMQRAGRLILKEFGPQSWIKGGHLK GGAKDFLFTK EQFVWESPRIQTCHTHGT
SP082 nucleotide (SEQ ID NO:139)
AATTGTACAATTAGAAAAAGATAGCAAATCAGACAAAGAACAAGTTGATAAACTATTTGAATCATTTGA TGCATCTTCAGATGAATCTATTTCTAAATTAAAAGAACTATCTGAAACTTCACTTAAAACCGATGCAGG TAAAGACTATCTTAATAACAAAGTCAAAGAATCATCTAAAGCAATTGTAGATTTTCATTTGCAAAAAGG TTTGGCTTATGATGTTAAAGATTCAGATGACAAATTTAAAGATAAAGCAACTCTTGAAACAAATGTAAA AGAAATTACAAAACAAATTGATTTTATCAAAAAAGTTGATGAAACTTTTAAACAAGAGAATTTGGAAGA AACTCTTAAATCTCTAAATGATCTTGTTGATAAATATCAAAAACAAATCGAACTTTTGAAGAAAGAAGA AGAAAAAGCTGCTGAAAAAGCTGCTGAAAAAGCAAAGGAATCTTCTAGTCAAAGTAATTCTTCTGGTAG TGCTTCTAATGAGTCTTATAATGGATCTTCCAATTCAAATGTAGATTATAGTTCATCTGAACAAACTAA TGGATATTCAAATAATTATGGCGGTCAAGATTATTCTGGTTCAGGAGATAGTTCAACAAATGGTGGATC ATCAGAACAATATTCATCTAGCAATTCAAACAGCGGAGCAAATAATGTCTACAGATATAAAGGCACTGG TGCTGACGGCTATCAAAGATACTACTACAAAGATCATAATAATGGAGATGTGTATGATGACGATGGAAA TTACCTTGGGAACTTTGGTGGCGGCATTGCAGAACCTAGTCAACGC
SP082 amino acid (SEQ ID NO:140)
IVQLEKDSKSDKEQVDKLFESFDASSDESISKLKELSETSLKT3AGKDYLNNKVKESSKAIVDFHLQKG LAYDVKDSDDKFKDKATLETNVKEITKQIDFIKKVDETFKQENLEΞTLKSLNDLVDKYQ QIEL KKEE EKAAEKAAEKAKESSSQSNΞSGSASNESYNGSSNSNVDYSSSEQTNGYSNNYGGQDYSGSGDSSTNGGS SEQYSSSNSNSGANNVYRYKGTGADGYQRYYYKDHNNGDVYDDDGNYLGNFGGGIAEPSQR
SP083 nucleotide (SEQ ID NO:141)
TCTGACCAAGCAAAAAGAAGCAGTCAATGACAAAGGAAAAGCAGCTGTTGTTAAGGTGGTGGAAAGCCA GGCAGAACTTTATAGCTTAGAAAAGAATGAAGATGCTAGCCTAAGAAAGTTACAAGCAGATGGACGCAT CACGGAAGAACAGGCTAAAGCTTATAAAGAATACAATGATAAAAATGGAGGAGCAAATCGTAAAGTCAA TGAT
SP083 amino acid (SEQ ID NO:142)
LTKQKEAVNDKGKAAWKWESQAELYSLEKNEDASLRKLQADGRITEEQAKAYKEYNDKNGGANRKVN D
SP084 nucleotide (SEQ ID NO: 143)
GTCCGGCTCTGTCCAGTCCACTTTTTCAGCGGTAGAGGAACAGATTTTCTTTATGGAGTTTGAAGAACT CTATCGGGAAACCCAAAAACGCAGTGTAGCCAGTCAGCAAAAGACTAGTCTGAACTTAGATGGGCAGAC GCTTAGCAATGGCAGTCAAAAGTTGCCAGTCCCTAAAGGAATTCAGGCCCCATCAGGCCAAAGTATTAC ATTTGACCGAGCTGGGGGCAATTCGTCCCTGGCTAAGGTTGAATTTCAGACCAGTAAAGGAGCGATTCG CTATCAATTATATCTAGGAAATGGAAAAATTAAACGCATTAAGGAAACAAAAAAT Table 1 79
SP084 amino acid (SEQ ID NO:144)
SGSVQSTFSAVEEQIFFMEFEELYRETQKRSVASQQKTSLNLDGQT SNGSQKLPVPKGIQAPSGQSIT FDRAGGNSS AKVEFQTSKGAIRYQLYLGNGKIKRIKETKN
SP085 nucleotide (SEQ ID NO:145)
GGGACAAATTCAAAAAAATAGGCAAGAGGAAGCAAAAATCTTGCAAAAGGAAGAAGTCTTGAGGGTAGC TAAGATGGCCCTGCAGACGGGGCAAAATCAGGTAAGCATCAACGGAGTTGAGATTCAGGTATTTTCTAG TGAAAAAGGATTGGAGGTCTACCATGGTTCAGAACAGTTGTTGGCAATCAAAGAGCCA
SP085 amino acid (SEQ ID NO:146)
GQIQKNRQEEAKILQKEEVLRVAKMALQTGQNQVSINGVEIQVFSSEKGLEVYHGSEQLLAIKEP
SP086 nucleotide (SEQ ID NO:147)
TCGCTACCAGCAACAAAGCGAGCAAAAGGAGTGGCTCTTGTTTGTGGACCAACTTGAGGTAGAATTAGA CCGTTCGCAGTTCGAAAAAGTAGAAGGCAATCGCCTATACATGAAGCAAGATGGCAAGGACATCGCCAT CGGTAAGTCAAAGTCAGATGATTTCCGTAAAACGAATGCTCGTGGTCGAGGTTATCAGCCTATGGTTTA TGGACTCAAATCTGTACGGATTACAGAGGACAATCAACTGGTTCGCTTTCATTTCCAGTTCCAAAAAGG CTTAGAAAGGGAGTTCATCTATCGTGTGGAAAAAGAAAAAAGT
SP086 amino acid (SEQ ID NO:148)
RYQQQSEQKE LLFVDQLEVELDRΞQFEKVEGNRLYMKQDGKDIAIGKSKSDDFRKTNARGRGYQPMVY GLKSVRITEDNQLVRFHFQFQKGLΞREFIYRVEKEKS
SP087 nucleotide (SEQ ID NO: 149)
GAACCGACAAGTCGCCCACTATCAAGACTATGCTTTGAATAAAGAAAAATTGGTTGCTTTTGCTATGGC TAAACGAACCAAAGATAAGGTTGAGCAAGAAAGTGGGGAACAGTTTTTTAATCTAGGTCAGGTAAGCTA TCAAAACAAGAAAACTGGCTTAGTGACGAGGGTTCGTACGGATAAGAGCCAATATGAGTTTCTGTTTCC TTCAGTCAAAATCAAAGAAGAGAAAAGAGATAAAAAGGAAGAGGTAGCGACCGATTCAAGCGAAAAAGT GGAGAAGAAAAAATCAGAAGAGAAGCCTGAAAAGAAAGAGAATTCA
SP087 amino acid (SEQ ID NO:150)
NRQVAHYQDYALNKEKLVAFAMAKRTKDKVEQESGEQFFNLGQVSYQNKKTGLVTRVRTDKSQYΞFLFP SVKIKEEKRDKKEEVATDSΞEKVEKKKSEEKPEKKENS
SP088 nucleotide (SEQ ID NO:151)
GGTTGTCGGCTGGCAATATATCCCGTTTCCATCTAAAGGTAGTACAATTGGTCCTTACCCAAATGGTAT CAGATTAGAAGGTTTTCCAAAGTCAGAGTGGTACTACTTCGATAAAAATGGAGTGCTACAAGAGTTTGT TGGTTGGAAAACATTAGAGATTAAAACTAAAGACAGTGTTGGAAGAAAGTACGGGGAAAAACGTGAAGA TTCAGAAGATAAAGAAGAGAAGCGTTATTATACGAACTATTACTTTAATCAAAATCATTCTTTAGAGAC AGGTTGGCTTTATGATCAGTCTAACTGGTATTATCTAGCTAAGACGGAAATTAATGGAGAAAACTACCT TGGTGGTGAAAGACGTGCGGGGTGGATAAACGATGATTCGACTTGGTACTACCTAGATCCAACAACTGG TATTATGCAAACAGGTTGGCAATATCTAGGTAATAAGTGGTACTACCTCCGTTCCTCAGGAGCAATGGC CACTGGCTGGTATCAGGAAGGTACCACTTGGTATTATTTAGACCACCCAAATGGCGATATGAAAACAGG TTGGCAAAACCTTGGGAACAAATGGTACTATCTCCGTTCATCAGGAGCTATGGCAACTGGTTGGTATCA AGATGGTTCAACTTGGTACTACCTAAATGCAGGTAATGGAGACATGAAGACAGGTTGGTTCCAGGTCAA TGGCAACTGGTACTATGCTTATAGCTCAGGTGCTTTGGCAGTGAATACGACCGTAGATGGCTATTCTGT CAACTATAATGGCGAATGGGTTCGG
SP088 amino acid (SEQ ID NO:152)
WG QYIPFPSKGSTIGPYPNGIRLEGFPKSE YYFDKNGVLQEFVGWKTLEIKTKDSVGRKYGEKRED SEDKEEKRYYTNYYFNQNHSLETG LYDQSNWYYLAKTEINGENYLGGERRAGWINDDST YYLDPTTG IMQTGWQYLGNK YYLRSSGAMATG YQEGTTtΑTYLDHPNGDMKTG QNLGNK YYLRSSGAMATGWYQ DGST YYLNAGNGDMKTGWFQλraGN YYAYSSGALAVNTTVDGYSVNYNGEWVR
SP089 nucleotide (SEQ ID NO:153)
GGCCAAATCAGAATGGGTAGAAGACAAGGGAGCCTTTTATTATCTTGACCAAGATGGAAAGATGAAAAG AAATGCTTGGGTAGGAACTTCCTATGTTGGTGCAACAGGTGCCAAAGTAATAGAAGACTGGGTCTATGA TTCTCAATACGATGCTTGGTTTTATATCAAAGCAGATGGACAGCACGCAGAGAAAGAATGGCTCCAAAT Table 1 80
TAAAGGGAAGGACTATTATTTCAAATCCGGTGGTTATCTACTGACAAGTCAGTGGATTAATCAAGCTTA TGTGAATGCTAGTGGTGCCAAAGTACAGCAAGGTTGGCTTTTTGACAAACAATACCAATCTTGGTTTTA CATCAAAGAAAATGGAAACTATGCTGATAAAGAATGGATTTTCGAGAATGGTCACTATTATTATCTAAA ATCCGGTGGCTACATGGCAGCCAATGAATGGATTTGGGATAAGGAATCTTGGTTTTATCTCAAATTTGA TGGGAAAATGGCTGAAAAAGAATGGGTCTACGATTCTCATAGTCAAGCTTGGTACTACTTCAAATCCGG TGGTTACATGACAGCCAATGAATGGATTTGGGATAAGGAATCTTGGTTTTATCTCAAATCTGATGGGAA AATAGCTGAAAAAGAATGGGTCTACGATTCTCATAGTCAAGCTTGGTACTACTTCAAATCCGGTGGTTA CATGACAGCCAATGAATGGATTTGGGATAAGGAATCTTGGTTTTACCTCAAATCTGATGGGAAAATAGC TGAAAAAGAATGGGTCTACGATTCTCATAGTCAAGCTTGGTACTACTTCAAATCTGGTGGCTACATGGC GAAAAATGAGACAGTAGATGGTTATCAGCTTGGAAGCGATGGTAAATGGCTTGGAGGAAAAACTACAAA TGAAAATGCTGCTTACTATCAAGTAGTGCCTGTTACAGCCAATGTTTATGATTCAGATGGTGAAAAGCT TTCCTATATATCGCAAGGTAGTGTCGTATGGCTAGATAAGGATAGAAAAAGTGATGACAAGCGCTTGGC TATTACTATTTCTGGTTTGTCAGGCTATATGAAAACAGAAGATTTACAAGCGCTAGATGCTAGTAAGGA CTTTATCCCTTATTATGAGAGTGATGGCCACCGTTTTTATCACTATGTGGCTCAGAATGCTAGTATCCC AGTAGCTTCTCATCTTTCTGATATGGAAGTAGGCAAGAAATATTATTCGGCAGATGGCCTGCATTTTGA TGGTTTTAAGCTTGAGAATCCCTTCCTTTTCAAAGATTTAACAGAGGCTACAAACTACAGTGCTGAAGA ATTGGATAAGGTATTTAGTTTGCTAAACATTAACAATAGCCTTTTGGAGAACAAGGGCGCTACTTTTAA GGAAGCCGAAGAACATTACCATATCAATGCTCTTTATCTCCTTGCCCATAGTGCCCTAGAAAGTAACTG GGGAAGAAGTAAAATTGCCAAAGATAAGAATAATTTCTTTGGCATTACAGCCTATGATACGACCCCTTA CCTTTCTGCTAAGACATTTGATGATGTGGATAAGGGAATTTTAGGTGCAACCAAGTGGATTAAGGAAAA TTATATCGATAGGGGAAGAACTTTCCTTGGAAACAAGGCTTCTGGTATGAATGTGGAATATGCTTCAGA CCCTTATTGGGGCGAAAAAATTGCTAGTGTGATGATGAAAATCAATGAGAAG
SP089 amino acid (SEQ ID NO:154)
AKSEWVEDKGAFYYLDQDGKMKRNA VGTSYVGATGAKVIEDWVYDSQYDA FYIKADGQHAEKEWLQI KGKDYYFKSGGYLLTSQ INQAYVNASGAKVQQGWLFDKQYQSWFYIKENGNYADKEWIFENGHYYYLK SGGYMAANE I DKES FYLKFDGKMAEKE VYDSHSQAWYYFKSGGYMTANEWIWDKESWFYLKSDGK IAEKEWVYDSHSQA YYFKSGGYMTANE I DKESWFYLKSDGKIAEKEWVYDΞHSQAWYYFKSGGYMA NETVDGYQLGSDGKWLGGKTTNENAAYYQWPVTANλArDSDGEKLSYISQGSW LDKDRKSDDKRLA ITISGLSGYMKTΞDLQALDASKDFIPYYESDGHRFYHYVAQNASIPVASHLSDMEVGKKYYSADGLHFD GFKLENPFLFKDLTEATNYSAEELDKVFSLLNINNSLLENKGATFKEAEEHYHINALY LAHSA ESNW GRSKIAKDfNNFFGITAYDTTPYLSAKTFDDVDKGILGATKWIKENYIDRGRTFLGNKASG NVEYASD PYWGEKIAΞVMMKINEK
SP090 nucleotide (SEQ ID NO:155)
ATTTGCAGATGATTCTGAAGGATGGCAGTTTGTCCAAGAAAATGGTAGAACCTACTACAAAAAGGGGGA TCTAAAAGAAACCTACTGGAGAGTGATAGATGGGAAGTACTATTATTTTGATCCTTTATCCGGAGAGAT GGTTGTCGGCTGGCAATATATACCTGCTCCACACAAGGGGGTTACGATTGGTCCTTCTCCAAGAATAGA GATTGCTCTTAGACCAGATTGGTTTTATTTTGGTCAAGATGGTGTATTACAAGAATTTGTTGGCAAGCA AGTTTTAGAAGCAAAAACTGCTACGAATACCAACAAACATCATGGGGAAGAATATGATAGCCAAGCAGA GAAACGAGTCTATTATTTTGAAGATCAGCGTAGTTATCATACTTTAAAAACTGGTTGGATTTATGAAGA GGGTCATTGGTATTATTTACAGAAGGATGGTGGCTTTGATTCGCGCATCAACAGATTGACGGTTGGAGA GCTAGCACGTGGTTGGGTTAAGGATTACCCTCTTACGTATGATGAAGAGAAGCTAAAAGCAGCTCCATG GTACTATCTAAATCCAGCAACTGGCATTATGCAAACAGGTTGGCAATATCTAGGTAATAGATGGTACTA CCTCCATTCGTCAGGAGCTATGGCAACTGGCTGGTATAAGGAAGGCTCAACTTGGTACTATCTAGATGC TGAAAATGGTGATATGAGAACTGGCTGGCAAAACCTTGGGAACAAATGGTACTATCTCCGTTCATCAGG AGCTATGGCAACTGGTTGGTATCAGGAAAGTTCGACTTGGTACTATCTAAATGCAAGTAATGGAGATAT GAAAACAGGCTGGTTCCAAGTCAATGGTAACTGGTACTATGCCTATGATTCAGGTGCTTTAGCTGTTAA TACCACAGTAGGTGGTTACTACTTAAACTATAATGGTGAATGGGTTAAG
SP090 amino acid (SEQ ID NO: 156)
VFADDSEG QFVQENGRTYYKKGDLKETYWRVIDGKYYYFDPLSGE WG QYIPAPHKGVTIGPSPRI EIALRPD FYFGQDGVLQEFVGKQVLEAKTATNTNKHHGEEYDSQAEKRVYYFEDQRSYHTLKTG IYE EGHWYYLQKDGGFDSRINRLTVGELARGWVKDYPLTYDEEKLKAAPWYYLNPATGIMQTGWQYLGNRWY YLHSSGAIVLATGWYKEGST YYLDAENGDMRTG QNLGNK YYLRSSGAMATGWYQESST YYLNASNGD KTG FQVNG YYAYDSGALAVNTTVGGYYLNYNGE V Table 1 81
SP091 nucleotide (SEQ ID NO: 157)
TGTCGCTGCAAATGAAACTGAAGTAGCAAAAACTTCGCAGGATACAACGACAGCTTCAAGTAGTTCAGA GCAAAATCAGTCTTCTAATAAAACGCAAACGAGCGCAGAAGTACAGACTAATGCTGCTGCCCACTGGGA TGGGGATTATTATGTAAAGGATGATGGTTCTAAAGCTCAAAGTGAATGGATTTTTGACAACTACTATAA GGCTTGGTTTTATATTAATTCAGATGGTCGTTACTCGCAGAATGAATGGCATGGAAATTACTACCTGAA ATCAGGTGGATATATGGCCCAAAACGAGTGGATCTATGACAGTAATTACAAGAGTTGGTTTTATCTCAA GTCAGATGGGGCTTATGCTCATCAAGAATGGCAATTGATTGGAAATAAGTGGTACTACTTCAAGAAGTG GGGTTACATGGCTAAAAGCCAATGGCAAGGAAGTTATTTCTTGAATGGTCAAGGAGCTATGATGCAAAA TGAATGGCTSCTATGATCCAGCCTATTCTGCTTATTTTTATCTAAAATCCGATGGAACTTATGCTAACC AAGAGTGGCAAAAAGTGGGCGGCAAATGGTACTATTTCAAGAAGTGGGGCTATATGGCTCGGAATGAGT GGCAAGGCAACTACTATTTGACTGGAAGTGGTGCCATGGCGACTGACGAAGTGATTATGGATGGTACTC GCTATATCTTTGCGGCCTCTGGTGAGCTCAAAGAAAAAAAAGATTTGAATGTCGGCTGGGTTCACAGAG ATGGTAAGCGCTATTTCTTTAATAATAGAGAAGAACAAGTGGGAACCGAACATGCTAAGAAAGTCATTG ATATTAGTGAGCACAATGGTCGTATCAATGATTGGAAAAAGGTTATTGATGAGAACGAAGTGGATGGTG TCATTGTTCGTCTAGGTTATAGCGGTAAAGAAGACAAGGAATTGGCGCATAACATTAAGGAGTTAAACC GTCTGGGAATTCCTTATGGTGTCTATCTCTATACCTATGCTGAAAATGAGACCGATGCTGAGAGTGACG CTAAACAGACCATTGAACTTATAAAGAAATACAATATGAACCTGTCTTACCCTATCTATTATGATGTTG AGAATTGGGAATATGTAAATAAGAGCAAGAGAGCTCCAAGTGATACAGGCACTTGGGTTAAAATCATCA ACAAGTACATGGACACGATGAAGCAGGCGGGTTATCAAAATGTGTATGTCTATAGCTATCGTAGTTTAT TACAGACGCGTTTAAAACACCCAGATATTTTAAAACATGTAAACTGGGTAGCGGCCTATACGAATGCTT TAGAATGGGAAAACCCTCATTATTCAGGAAAAAAAGGTTGGCAATATACCTCTTCTGAATACATGAAAG GAATCCAAGGGCGCGTAGATGTCAGCGTTTGGTAT
SP091 amino acid (SEQ ID NO:158)
VAANETEVAKTΞQDT^TASSSSEQNQSSNKTQTSAEVQTNAAAH DGDYYVKDDGSKAQΞEWIFDNYYK AWFYINSDGRYSQNE HGNYYLKSGGYMAQNEWIYDSNYKSWFYLKSDGAYAHQE QLIGNK YYFKKW GYMAKSQWQGSYFLNGQGAMMQNEWLYDPAYSAYFYLKSDGTYANQEWQKVGGK YYFKK GYMARNEW QGNYYLTGSGAMATDEVIMDGTRYIFAASGELKEKKDLNVG VHRDGKRYFFNNREEQVGTEHAKKVID ISEHNGRINDWKKVIDENEVDGVIVRLGYΞGKEDKELAHNIKELNRLGIPYGVYLYTYAENETDAESDA KQTIELIKKYNMNLSYPIYYDVE WEYVNKSKRAPSDTGTWVKIINKYMDTMKQAGYQNVYVYSYRSLL QTR KHPDILKHV WVAAYTNALE ENPHYΞGKKGWQYTSSEYMKGIQGRVDVSV Y
SP092 nucleotide (SEQ ID NO: 159)
TACGTCTCAGCCTACTTTTGTAAGAGCAGAAGAATCTCCACAAGTTGTCGAAAAATCTTCATTAGAGAA GAAATATGAGGAAGCAAAAGCAAAAGCTGATACTGCCAAGAAAGATTACGAAACGGCTAAAAAGAAAGC AGAAGACGCTCAGAAAAAGTATGAAGATGATCAGAAGAGAACTGAGGAGAAAGCTCGAAAAGAAGCAGA AGCATCTCAAAAATTGAATGATGTGGCGCTTGTTGTTCAAAATGCATATAAAGAGTACCGAGAAGTTCA AAATCAACGTAGTAAATATAAATCTGACGCTGAATATCAGAAAAAATTAACAGAGGTCGACTCTAAAAT AGAGAAGGCTAGGAAAGAGCAACAGGACTTGCAAAATAAATTTAATGAAGTAAGAGCAGTTGTAGTTCC TGAACCAAATGCGTTGGCTGAGACTAAGAAAAAAGCAGAAGAAGCTAAAGCAGAAGAAAAAGTAGCTAA GAGAAAATATGATTATGCAACTCTAAAGGTAGCACTAGCGAAGAAAGAAGTAGAGGCTAAGGAACTTGA AATTGAAAAACTTCAATATGAAATTTCTACTTTGGAACAAGAAGTTGCTACTGCTCAACATCAAGTAGA TAATTTGAAAAAACTTCTTGCTGGTGCGGATCCTGATGATGGCACAGAAGTTATAGAAGCTAAATTAAA AAAAGGAGAAGCTGAGCTAAACGCTAAACAAGCTGAGTTAGCAAAAAAACAAACAGAACTTGAAAAACT TCTTGACAGCCTTGATCCTGAAGGTAAGACTCAGGATGAATTAGATAAAGAAGCAGAAGAAGCTGAGTT GGATAAAAAAGCTGATGAACTTCAAAATAAAGTTGCTGATTTAGAAAAAGAAATTAGTAACCTTGAAAT ATTACTTGGAGGGGCTGATNCTGAAGATGATACTGCTGCTCTTCAAAATAAATTAGCTACTAAAAAAGC TGAATTGGAAAAAACTCAAAAAGAATTAGATGCAGCTCTTAATGAGTTAGGCCCTGATGGAGATGAAGA AGAAACTCCAGCGCCGGCTCCTCAACCAGAGCAACCAGCTCCTGCACCAAAACCAGAGCAACCAGCTCC AGCTCCAAAACCAGAGCAACCAGCTCCTGCACCAAAACCAGAGCAACCAGCTCCAGCTCCAAAACCAGA GCAACCAGCTCCAGCTCCAAAACCAGAGCAACCAGCTAAGCCGGAGAAACCAGCTGAAGAGCCTACTCA ACCAGAAAAACCAGCCACTCCAAAAACAGGCTGGAAACAAGAAAACGGTATGTGGTATTTCTACAATAC TGATGGTTCAATGGCAATAGGTTGGCTCCAAAACAACGGTTCATGGTACTACCTAAACGCTAACGGCGC TATGGCAACAGGTTGGGTGAAAGATGGAGATACCTGGTACTATCTTGAAGCATCAGGTGCTATGAAAGC AAGCCAATGGTTCAAAGTATCAGATAAATGGTACTATGTCAACAGCAATGGCGCTATGGCGACAGGCTG GCTCCAATACAATGGCTCATGGTACTACCTCAACGCTAATGGTGATATGGCGACAGGATGGCTCCAATA CAACGGTTCATGGTATTACCTCAACGCTAATGGTGATATGGCGACAGGATGGGCTAAAGTCAACGGTTC ATGGTACTACCTAAACGCTAACGGTGCTATGGCTACAGGTTGGGCTAAAGTCAACGGTTCATGGTACTA Table 1 82
CCTAAACGCTAACGGTTCAATGGCAACAGGTTGGGTGAAAGATGGAGATACCTGGTACTATCTTGAAGC ATCAGGTGCTATGAAAGCAAGCCAATGGTTCAAAGTATCAGATAAATGGTACTATGTCAATGGCTTAGG TGCCCTTGCAGTCAACACAACTGTAGATGGCTATAAAGTCAATGCCAATGGTGAATGGGTT
SP092 amino acid (SEQ ID NO: 160)
TSQPTFVRAEESPQWEKSSLEKKYEEAKAKADTAKKDYETAKKKAEDAQKKYEDDQKRTEEKARKEAE ASQKLNDVALWQNAYKEYREVQNQRSKYKΞDAEYQKKLTΞVDSKIEKARKEQQDLQNKFNEVRAVWP EPNALAETKKKAEEAKAEEKVAKRKYDYATLKVALAKKEVEAKELEIEK QYEISTLEQEVATAQHQVD NLKKLLAGADPDDGTEVIEAKLKKGEAELNAKQAELAKKQTE EKLLDSLDPEGKTQDE DKEAEEAEL DKKADELQNKVADLEKEISN EI LGGADXEDDTAALQNKLATKKAELEKTQKELDAALNELGPDGDEE ETPAPAPQPEQPAPAPKPEQPAPAPKPEQPAPAPKPEQPAPAPKPEQPAPAPKPEQPAKPEKPAEEPTQ PEKPATPKTG KQENGMYFYNTDGSMAIGWLQMSIGSWYYLNANGARATGWVKDGDT YYLEASGAMKA SQWFKVSDKWYYVNSNGAMATGWLQYNGS YYLNANGDMATG LQYNGSVIΓΓLNANGDMATGAKVNGS YYLNANGAMATGWAKVNGSWYYLNANGSMATGWVKDGDTWYYLEASGAMKASQWFKVSDK YYVNGLG ALAVNTTVDGYKVNANGEWV
P093 nucleotide (SEQ ID NO: 161)
TGGACAGGTGAAAGGTCATGCTACATTTGTGAAATCCATGACAACTGAAATGTACCAAGAACAACAGAA CCATTCTCTCGCCTACAATCAACGCTTGGNTTCGCAAAATCGCATTGTAGATCCTTTTTTGGCGGAGGG ATATGAGGTCAATTACCAAGTGTCTGACGACCCTGATGCAGTCTATGGTTACTTGTCTATTCCAAGTTT GGAAATCATGGAGCCGGTTTATTTGGGAGCAGATTATCATCATTTAGGGATGGGCTTGGCTCATGTGGA TGGTACACCGCTGCCTCTGGATGGTACAGGGATTCGCTCAGTGATTGCTGGGCACCGTGCAGAGCCAAG CCATGTCTTTTTCCGCCATTTGGATCAGCTAAAAGTTGGAGATGCTCTTTATTATGATAATGGCCAGGA AATTGTAGAATATCAGATGATGGACACAGAGATTATTTTACCGTCGGAATGGGAAAAATTAGAATCGGT TAGCTCTAAAAATATCATGACCTTGATAACCTGCGATCCGATTCCTACCTTTAATAAACGCTTATTAGT GAATTTTGAACGAGTCGCTGTTTATCAAAAATCAGATCCACAAACAGCTGCAGTTGCGAGGGTTGCTTT TACGAAAGAAGGACAATCTGTATCGCGTGTTGCAACCTCTCAATGGTTG
SP093 amino acid (SEQ ID NO:162)
GQVKGHATFVKSMTTEMYQEQQNHSLAYNQRLXSQNRIVDPFLAEGYEVNYQVSDDPDAVYGYLSIPSL EIMEPVYLGADYHHLGMG AHVDGTPLPLDGTGIRSVIAGHRAEPSHVFFRHLDQLKVGDA YYDNGQE IVEYQMMDTEIILPSE EKLESVSSKNI TLITCDPIPTFNKRLLVNFERVAVYQKSDPQTAAVARVAF TKEGQSVSRVATSQWL
SP094 nucleotide (SEQ ID NO:163)
GATTGCTCCTTTGAAGGATTTGAGAGAAACCATGTTGGAAATTGCTTCTGGTGCTCAAAATCTTCGTGC CAAGGAAGTTGGTGCCTATGAACTGAGAGAAGTAACTCGCCAATTTAATGCTATGTTGGATCAGATTGA TCAGTTGATGGTAGCTATTCGTAGCCAGGAAGAAACGACCCGTCAGTACCAACTTCAAGCCCTTTCGAG CCAGATTAATCCACATTTCCTCTATAACACTTTGGACACCATCATCTGGATGGCTGAATTTCATGATAG TCAGCGAGTGGTGCAGGTGACCAAGTCCTTGGCAACCTATTTCCGCTTGGCGCTCAATCAAGGCAAGGA CTTGATTTGTCTCTCTGACGAAATCAATCATGTCCGCCAGTATCTCTTTATCCAGAAACAACGCTATGG AGATAAGCTGGAATACGAAATTAATGAAAATGTTGCCTTTGATAATTTAGTCTTACCCAAGCTGGTCCT ACAACCCCTTGTAGAAAATGCTCTTTACCATGGCATTAAGGAAAAGGAAGGTCAGGGCCATATTAAACT TTCTGTCCAGAAACAGGATTCGGGATTGGTCATCCGTATTGAGGATGATGGCGTTGGCTTCCAAGATGC TGGTGATAGTAGTCAAAGTCAACTCAAACGTGGGGGAGTTGGTCTTCAAAATGTCGATCAACGGCTCAA ACTTCATTTTGGAGCCAATTACCATATGAAGATTGATTCTAGACCCCAAAAAGGGACGAAAGTTGAAAT ATATATAAATAGAATAGAAACTAGC
SP094 amino acid (SEQ ID NO:164)
IAPLKDLRETM EIAΞGAQNLRAKEVGAYELREVTRQFNAMLDQIDQLMVAIRSQEETTRQYQLQALSS QINPHF YNTLDTIIWMAEFHDSQRWQVTKSLATYFRLALNQGKDLICLSDEINHVRQYLFIQKQRYG DK EYEINE^ AFDN VLPKLV QPLVENA YHGIKEKEGQGHIKLSVQKQDSG VIRIEDDGVGFQDA GDSSQSQLKRGGVGLQNVDQRLKLHFGANYHMKIDSRPQKGTKVEIYINRIETΞ
SP095 nucleotide (SEQ ID NO: 165)
TAGGTCATATGGGACTTTTTTTCTACAACAAAATAGGCTCCATAATATCTATAAGGGATTTACCCACTA CAAATATTATAGAGCCGAAAATTCACATCTAATATATGCAGACTACTTTGAAATGAAATTAAAAAAATT ATTAAAGGATGACACAAAAGTTTTTGAAAAATCTACATTCAAATTTGTAGAAGGATATAAAATATACCT Table 1 83
GACAGAATCTAAAGAATCTGGAATTAAACAAATGGACAATGTCATAAAATATTTTGAGTTTATTGAATC TAAAAGTATTGCTTTATATTTTCAAAAACGATTAAATGAGCTGATAGAT
SP095 amino acid (SEQ ID NO: 166)
RSYGTFFLQQNRLHNIYKGFTHYKYYRAENSHLIYADYFEMKLKKLLKDDTKVFEKΞTFKFVEGYKIYL TESKEΞGIKQMDNVIKYFEFIESKSIALYFQKRLNE ID
SP096 nucleotide (SEQ ID NO:167)
CAACGTTGAGAATTATTTGCGAATGTGTTTGGATAGCATTCAGAATCAGACGTATCAAAATTTTGAGTG TTTATTAATCAATGATGGCTCTCCAGATCATTCATCCAAAATATGTGAAGAATTTGTAGAGAAAGATTC TCGTTTCAAATATTTTGAGAAAGCAAACGGCGGTCTTTCATCAGCTCGTAACCTAGGTATTGAATGTTC GGGGGGGGGCGTACATTACTTTTGTAGACTC
SP096 amino acid (SEQ ID NO:168)
NVENYLRMCLDSIQNQTYQNFECLLINDGSPDHSSKICEEFVEKDSRFKYFEKANGGLS3ARNLGIECΞ GGGVHYFCRL
SP097 nucleotide (SEQ ID NO:169)
CTACTATCAATCAAGTTCTTCAGCCATTGAGGCCACCATTGAGGGCAACAGCCAAACGACCATCAGCCA GACTAGCCACTTTATTCAGTCTTATATCAAAAAACTAGAAACCACCTCGACTGGTTTGACCCAGCAGAC GGATGTTCTGGCCTATGCTGAGAATCCCAGTCAAGACAAGGTCGAGGGAATCCGAGATTTGTTTTTGAC CATCTTGAAGTCAGATAAGGACTTGAAAACTGTTGTGCTGGTGACCAAATCTGGTCAGGTCAT'TTCTAC AGATGACAGTGTGCAGATGAAAACTTCCTCTGATATGATGGCTGAGGATTGGTACCAAAAGGCCATTCA TCAGGGAGCTATGCCTGTTTTGACTCCAGCTCGTAAATCAGATAGTCAGTGGGTCATTTCTGTCACTCA AGAACTTGTTGATGCAAAGGGAGCCAATCTTGGTGTGCTTCGTTTGGATATTTCTTATGAAACTCTGGA AGCCTATCTCAATCAACTCCAGTTGGGGCAGCAGGGCTTTGCCTTCATTATCAATGAAAACCATGAATT TGTCTACCATCCTCAACACACAGTTTATAGTTCGTCTAGCAAAATGGAGGCTATGAAACCCTACATCGA TACAGGTCAGGGTTATACTCCTGGTCACAAATCCTACGTCAGTCAAGAGAAGATTGCAGGAACTGATTG GACGGTGCTTGGCGTGTCATCATTGGAAAAGTTAGACCAGGTTCGGAGTCAG
SP097 amino acid (SEQ ID NO: 170)
YYQSSSSAIEATIEGNSQTTISQTSHFIQSYIK LETTSTGLTQQTDV AYAENPSQDKVEGIRDLFLT ILKSDKDLKTWLVTKSGQVIΞTDDΞVQMKTSSDMMAED YQKAIHQGAMPVLTPARKSDSQWVISVTQ ELVDAKGAN GV RLDISYETLEAY NQLQLGQQGFAFIINENHEFVYHPQHTVYSSSSKMEAMKPYID TGQGYTPGHKSYVSQEKIAGTDWTV GVSSLEKLDQVRSQ
SP098 nucleotide (SEQ ID NO:171)
GACAAAAACATTAAAACGTCCTGAGGTTTTATCACCTGCAGGGACTTTAGAGAAGCTAAAGGTAGCTGT TCAGTATGGAGCAGATGCTGTCTTTATCGGTGGTCAGGCCTATGGTCTTCGTAGCCGTGCGGGAAACTT TACTTTCGAACAGATGGAAGAAGGCGTGCAGTTTGCGGCCAAGTATGGTGCCAAGGTCTATGTAGCGGC TAATATGGTTATGCACGAAGGAAATGAAGCTGGTGCTGGTGAGTGGTTCCGTAAACTGCGTGATATCGG GATTGCAGCAGTTATCGTATCTGACCCAGCCTTGATTATGATTGCAGTGACTGAAGCACCAGGCCTTGA AATCCACCTTTCTACCCAAGCCAGTGCCACTAACTATGAAACCCTTGAGTTCTGGAAAGAGCTAGGCTT GACTCGTGTCGTTTTAGCGCGTGAGGTTTCAATGGAAGAATTAGCTGAGATCCGCAAACGTACAGATGT TGAAATTGAAGCCTTTGTCCATGGAGCTATGTGTATTTCATACTCTGGACGTTGTACTCTTTCAAACCA CATGAGTATGCGTGATGCCAACCGTGGTGGATGTTCTCAGTCATGCCGTTGGAAATACGACCTTTACGA TATGCCATTTGGGAAAGAACGTAAGAGTTTGCAGGGTGAGATTCCAGAAGAATTTTCAATGTCAGCCGT TGACATGTCTATGATTGACCANATTCCAGATATGATTGAAAATGGTGTGGACAGTCTAAAAATCGAAGG ACGTATGNAGTCTATTCACTANGTATCAACAGTAACCAACTGCTACAAGGCGGCTGTGGATGCCTATCT TGAAAGTCCTGAAAAGTTTGAAGCTATCAAACAAGACTTGGTGGACGAGATGTGGAAGGTTGCCCAACG TGAACTGGCTACAGGATTTTACTATGGTACACCATCTGAAAATGAGCAGTTGTTTGGTGCTCGTCGTAA AATCCCTGAGTACAAGTTTGTCGCTGAAGTGGTTTCTTATGATGATGCGGCACAAACAGCAACTATTCG TCAACGAAACGTCATTAACGAAGGGGACCAAGTTGAGTTTTATGGTCCAGGTTTCCGTCATTTTGAAAC CTATATTGAAGATTTGCATGATGCTAAAGGCAATAAAATCGACCGCGCTCCAAATCCAATGGAACTATT GACTATTAAAGTCCCACAACCTGTTCAATCAGGAGACATGGTTCGAGCTCTTAAAGAGGGGCTTATCAA TCTTTATAAGGAAGATGGAACCAGCGTCACAGTTCGTGCT Table 1 84
SP098 amino acid (SEQ ID NO: 172)
TKTLKRPEVLSPAGTLEKLKVAVQYGADAVFIGGQAYGLRSRAGNFTFEQMEEGVQFAAKYGAKVYVAA NMVMHEGNEAGAGEWFRKLRDIGIAAVIVSDPALIMIAVTEAPGLEIHLSTQASATNYETLEFWKELG TRWLAREVSMEE AEIRKRTDVEIΞAFVHGAMCISYSGRCTLSNHMSMRDANRGGCSQSCRWKYDLYD MPFGKERKSLQGΞIPEEFSMSAVD S IDXIPDMIENGVDSLKIEGRMXSIHXVSTVTNCYKAAVDAYL ESPEKFEAIKQDLVDEM KVAQRELATGFYYGTPSENEQLFGARRKIPEYKFVAEWSYDDAAQTATIR QRNVINEGDQVΞFYGPGFRHFETYIEDLHDAKGNKIDRAPNPMELLTIKVPQPVQSGDMVRALKEGLIN LYKEDGTSVTVRA
SP099 nucleotide (SEQ ID NO:173)
TTCTCAGGAGACCTTTAAAAATATCACCAATAGCTTCTCCATGCAAATCAATCGTCGCGTCAACCAAGG AACGCCTCGTGGTGCTGGGAATATCAAGGGTGAAGACATCAAAAAAATCACCGAAAACAAGGCCATTGA GTCTTATGTCAAACGTATCAACGCTATCGGAGATTTGACTGGATATGACCTGATTGAAACGCCAGAAAC CAAGAAGAATCTCACTGCTGATCGTGCCAAGCGTTTTGGAAGTAGCTTGATGATTACAGGTGTCAATGA CTCCTCTAAAGAAGACAAGTTTGTCTCTGGTTCTTATAAACTAGTCGAAGGAGAGCACTTAACCAACGA CGACAAGGATAAAATCCTCTTGCACAAGGACTTGGCAGCCAAACACGGCTGGAAAGTAGGGGACAAGGT TAAACTGGACTCTAATATCTACGATGCAGATAATGAAAAAGGAGCCAAGGAAACAGTTGAAGTGACAAT CAAGGGACTCTTTGATGGTCATAATAAGTCAGCAGTAACCTACTCACAAGAACTTTACGAAAACACAGC TATTACAGACATTCACACTGCTGCAAAACTTTATGGATACACAGAAGACACAGCCATTTATGGGGACGC AACCTTCTTTGTAACAGCAGACAAGAACTTGGATGATGTTATGAAAGAGTTGAATGGCATCAGTGGTAT CAACTGGAAGAGCTACACACTCGTCAAGAGCTCCTCTAACTACCCAGCTCTTGAGCAATCTATCTCTGG TATGTACAAGATGGCCAAC
SP099 amino acid (SEQ ID NO:174)
SQETFKNITNSFSMQINRRVNQGTPRGAGNIKGEDIKKITΞNKAIESYVKRINAIG3LTGYDLIETPET KKNLTADRAKRFGSSL ITGVNDSSKEDKFVSGSYKLVEGEHLTNDDKDKILLHKDLAAKHGWKVGDKV KLDSNIYDADNEKGAKETVEVTIKGLFDGHNKSAVTYSQELYENTAITDIHTAAKLYGYTΞDTAIYGDA TFFVTADKNLDDVMKELNGISGINWKSYTLVKSSSNYPALEQSISG YKMAN
SP100 nucleotide (SEQ ID NO: 175)
AGTAAATGCGCAATCAAATTCATTAATATTAATAGATGAACCTGAAATCTCACTTCATCCGAGTGCAAT CTATAAATTTAAAGAGTTTTTACTTCAAGAGTGTTTAAATAAAAAACATCAAATTATTATCACTACACA TTCTACACAACTTATAAAAGATTTTCCTAGAGAAGCCGTGAAACTTTTAGTGAAAAACGGAGAAAAGGT AGATGTTATTGAAAATATTGATTATCAGGATGCATTTTTTGAATTAGGTGATGTGTATCATTCTAGGAA GATGATTTATGTTGAAGATAGACTAGCTAAATATATTCTAGAGTTTGTTATCACTCATTCAGGTAGTGA GAATCTTAAACAGAATTTAGTAGTGAGATATATTCCTGGTGGAGCAAATCAAATAATTTGTAATAATAT TTTAAACTCATCGTATTTAGATTCCGATAACCATTATTTTTGGCTTGATGGAGATCAAAACACTAATGT TAGTGAATCAAATAATTTAATGAACTATCTTGAAAATGGTGTTGTTATATCAGATAAAATTCCTGAATC AGATAATAAAAATCTTGATGATATTATAAAATTGATAANGGGATGTCCAATTAAATTTAATGTTTCAGG TAATAAAGGGCAAAAAAATAATATTGAATTAATTGCGAAACAAAGAAGCTTTATAGATTATTGGGCTAA ATAC
SP100 amino acid (SEQ ID NO:176)
VNAQSNSLILIDEPEISLHPSAIYKFKEFLLQECLNKKHQIIITTHSTQLIKDFPREAVKLLVKNGEKV DVIENIDYQDAFFELGDVYHSRKMIYVEDRLAKYILEFVITHSGΞENLKQNLWRYIPGGANQIICNNI LNSSYLDSDNHYF LDGDQNTNVSESNNLMNYLENGWISDKIPESDNKNLDDIIKLIXGCPIKFNVSG NKGQKNNIELIAKQRSFIDY AKY
SP101 nucleotide (SEQ ID NO:177)
TTACCGCGTTCATCAAGATGTCAAACAAGTCATGACCTATCAACCCATGGTGCGAGAAATATTGAGTGA ACAAGACACCCCAGCAAACGAAGAGCTTGTGCTTGCTATGATTTATACTGAAACAAAAGGAAAAGAAGG CGATGTTATGCAGTCTAGTGAGTCTGCAAGTGGTTCCACCAACACCATCAATGATAATGCCTCTAGCAT TCGGCAAGGCATTCAAACTCTGACAGGCAATCTCTATCTGGCGCAGAAGAAGGGGGTAGATATCTGGAC AGCTGTTCAAGCCTATAATTTTGGACCTGCCTATATCGATTTTATCGCCCAAAATGGCAAGGAAAATAC CCTGGCTCTAGCCAAACAGTACTCTCGTGAGACTGTTGCCCCCTTGCTTGGTAATAGGACTGGAAAGAC TTATAGTTATATTCACCCCATTTCCATTTTTCACGGTGCTGAACTCTATGTAAATGGAGGAAACTATTA TTATTCTAGACAGGTACGACTTAACCTTTACATCATCAAATGTTTCACTCTCTTTTCAACATCTGGC Table 1 85
SP101 amino acid (SEQ ID NO: 178)
YRVHQDVKQVMTYQPMVREILSEQDTPANEELVLAMIYTETKGKEGDVMQSSESASGSTNTINDNASSI RQGIQTLTGNLY AQKKGVDIWTAVQAYNFGPAYIDFIAQNGKENTLALAKQYSRETVAP LGNRTGKT YSYIHPISIFHGAΞ YVNGGNYY^SRQVRLNLYIIKCFTLFSTΞG
SP102 nucleotide (SEQ ID NO:179)
GTGGATGGGCTTTAACTATCTTCGTATTCGCCGTGCGGCTAAAATTGTGGACAATGAGGAGTTTGAAGC CTTGATTCGTACGGGTCAATTGATTGATTTGCGCGACCCAGCAGAATTCCACAGAAAACATATCCTTGG TGCACGCAATATTCCTTCAAGTCAGTTGAAAACTAGTCTTGCAGCCCTTCGTAAAGATAAACCTGTCCT TCTCTACGAAAACCAACGTGCGCAACGAGTTACAAATGCAGCTCTTTACTTGAAAAAACAAGGTTTTTC TGAGATTTATATCCTTTCTTATGGCTTGGATTCTTGGAAAGGGAAAGTGAAGACTAGC
SP102 amino acid (SEQ ID NO: 180)
WMGFNY RIRRAAKIVDNEEFΞALIRTGQLIDLRDPAEFHRKHILGARNIPSSQLKTSLAALRKDKPVL LYENQRAQRVTNAALYLKKQGFSΞIYILSYGLDS KGKVKTS
SP103 nucleotide (SEQ ID NO: 181)
ACTAAACCAGCATCGTTCGCAGGAAAATAAGGACAATAATCGTGTCTCTTATGTGGATGGCAGCCAGTC AAGTCAGAAAAGTGAAAACTTGACACCAGACCAGGTTAGCCAGAAAGAAGGAATTCAGGCTGAGCAAAT TGTAATCAAAATTACAGATCAGGGCTATGTAACGTCACACGGTGACCACTATCATTACTATAATGGGAA AGTTCCTTATGATGCCCTCTTTAGTGAAGAACTCTTGATGAAGGATCCAAACTATCAACTTAAAGACGC TGATATTGTCAATGAAGTCAAGGGTGGTTATATCATCAAGGTCGATGGAAAATATTATGTCTACCTGAA AGATGCAGCTCATGCTGATAATGTTCGAACTAAAGATGAAATCAATCGTCAAAAACAAGAACATGTCAA AGATAATGAGAAGGTTAACTCTAATGTTGCTGTAGCAAGGTCTCAGGGACGATATACGACAAATGATGG TTATGTCTTTAATCCAGCTGATATTATCGAAGATACGGGTAATGCTTATATCGTTCCTCATGGAGGTCA CTATCACTACATTCCCAAAAGCGATTTATCTGCTAGTGAATTAGCAGCAGCTAAAGCACATCTGGCTGG AAAAAATATGCAACCGAGTCAGTTAAGCTATTCTTCAACAGCTAGTGACAATAACACGCAATCTGTAGC AAAAGGATCAACTAGCAAGCCAGCAAATAAATCTGAAAATCTCCAGAGTCTTTTGAAGGAACTCTATGA TTCACCTAGCGCCCAACGTTACAGTGAATCAGATGGCCTGGTCTTTGACCCTGCTAAGATTATCAGTCG TACACCAAATGGAGTTGCGATTCCGCATGGCGACCATTACCACTTTATTCCTTACAGCAAGCTTTCTGC CTTAGAAGAAAAGATTGCCAGAATGGTGCCTATCAGTGGAACTGGTTCTACAGTTTCTACAAATGCAAA ACCTAATGAAGTAGTGTCTAGTCTAGGCAGTCTTTCAAGCAATCCTTCTTCTTTAACGACAAGTAAGGA GCTCTCTTCAGCATCTGATGGTTATATTTTTAATCCAAAAGATATCGTTGAAGAAACGGCTACAGCTTA TATTGTAAGACATGGTGATCATTTCCATTACATTCCAAAATCAAATCAAATTGGGCAACCGACTCTTCC AAACAATAGTCTAGCAACACCTTCTCCATCTCTTCCAATCAATCCAGGAACTTCACATGAGAAACATGA AGAAGATGGATACGGATTTGATGCTAATCGTATTATCGCTGAAGATGAATCAGGTTTTGTCATGAGTCA CGGAGACCACAATCATTATTTCTTCAAGAAG
SP103 amino acid (SEQ ID NO: 182)
LNQHRSQENKDNNRVSYVDGSQSSQKSENLTPDQVSQKEGIQAEQIVIKITDQGYVTSHGDHYHYYNGK VPYDA FSEELLMKDPNYQLKDADIVNEVKGGYIIKVDGKYYVY KDAAHADNVRTKDEINRQKQEHVK DNEKVNSNVAVARSQGRYTTNDGYVFNPADIIEDTGNAYIVPHGGHYHYIPKSDLSASELAAAKAHLAG KNMQPSQLSYSSTASDNNTQSVAKGSTSKPANKSENLQSLLKE YDSPSAQRYSESDGLVFDPAKIISR TPNGVAIPHGDKYHFIPYSK SALEEKIARMVPIΞGTGSTVSTNAKPNEWSSLGSLSSNPSSLTTSKE LSSASDGYIFNPKDIVEETATAYIVRHGDHFHYIPKSNQIGQPTLPNNSLATPSPSLPINPGTSHEKHE EDGYGFDANRIIAEDESGFVMSHGDHNHYFFKK
SP105 nucleotide (SEQ ID NO:183)
TGACTACCTTGAAATCCCACTTTACAGCTATCTTGGTGGATTCAACACTAAAGTTCTTCCAACTCCAAT GATGAACATCATCAACGGTGGTTCTCACTCTGACGCTCCAATCGCTTTCCAAGAGTTCATGATCTTGCC AGTTGGTGCGCCAACATTTAAAGAAGCCCTTCGTTACGGTGCTGAAATCTTCCACGCTCTTAAGAAAAT CCTTAAATCACGTGGTTTGGAAACTGCCGTAGGTGACGAAGGTGGATTCGCTCCTCGTTTCGAAGGAAC TGAAGATGGTGTTGAAACTATCCTTGCTGCGATTGAAGCTGCTGGATATGTACCAGGTAAAGACGTATT TATCGGATTTGACTGTGCTTCATCAGAATTCTACGATAAAGAACGTAAAGTTTACGACTACACTAAATT TGAAGGTGAAGGTGCTGCTGTTCGTACATCTGCAGAACAAATCGACTACCTTGAAGAATTGGTTAACAA ATACCCAATCATCACTATTGAAGATGGTATGGATGAAAACGACTGGGATGGTTGGAAAGCTCTTACTGA ACGTCTTGGTAAGAAAGTACAACTTGTTGGTGACGACTTCTTCGTAACAAACACTGACTACCTTGCACG Table 1 86
TGGTATCCAAGAAGGTGCTGCTAACTCAATCCTTATCAAAGTTAACCAAATCGGTACTCTTACTGAAAC TTTTGAAGCTATCGAAATGGCTAAAGAAGCTGGTTACACTGCTGTTGTATCACACCGTTCAGGTGAAAC TGAAGATTCAACAATCGCTGATATTGCAGTTGCAACTAACGCAGGACAAATCAAGACTGGTTCACTTTC ACGTACAGACCGCATCGCTAAATACAACCAATTGCTTCGTATCGAAGACCAACTTGGTGAAGTAGCTGA ATATCGTGGATTGAAATCATTCTACAACCTTAAAAAA
SP105 amino acid (SEQ ID NO:184)
DYLEIPLYSYLGGFNTKVLPTPMMNIINGGSHSDAPIAFQEFMILPVGAPTFKEALRYGAEIFHALKKI LKSRG ETAVGDEGGFAPRFΞGTΞDGVETILAAIEAAGYVPGKDVFIGFDCASSEFYDKERKVY3YTKF EGEGAAVRTSAEQIDYLEELVNKYPIITIEDGMDENDWDG KALTERLGKKVQLVGDDFFVTNTDYLAR GIQEGAANSILIKVNQIGTLTΞTFEAIEMAKEAGYTAWSHRSGΞTEDSTIADIAVATNAGQIKTGSLS RTDRIAKYNQLLRIEDQLGEVAEYRGLKSFYN KK
SP106 nucleotide (SEQ ID NO:185)
TCGTATCTTTTTTTGGAGCAATGTTCGCGTAGAAGGACATTCCATGGATCCGACCCTAGCGGATGGCGA AATTCTCTTCGTTGTAAAACACCTTCCTATTGACCGTTTTGATATCGTGGTGGCCCATGAGGAAGATGG CAATAAGGACATCGTCAAGCGCGTGATTGGAATGCCTGGCGACACCATTCGTTACGAAAATGATAAACT CTACATCAATGACAAAGAAACGGACGAGCCTTATCTAGCAGACTATATCAAACGCTTCAAGGATGACAA ACTCCAAAGCACTTACTCAGGCAAGGGCTTTGAAGGAAATAAAGGAACTTTCTTTAGAAGTATCGCTCA AAAAGCTCAAGCCTTCACAGTTGATGTCAACTACAACACCAACTTTAGCTTTACTGTTCCAGAAGGAGA ATACCTTCTCCTCGGAGATGACCGCTTGGTTTCGAGCGACAGCCGCCACGTAGGTACCTTCAAAGCAAA AGATATCACAGGGGAAGCTAAATTCCGCTTATGGCCAATCACCCGTATCGGAACATTT
SP106 amino acid (SEQ ID NO: 186)
RIFFWSNVRVEGHSMDPTLADGEILFWKHLPIDRFDIWAHEΞDGNKDIVKRVIGMPGDTIRYΞNDKL YINDKETDEPY ADYIKRFKDDKLQSTYSGKGFΞGNKGTFFRSIAQKAQAFTVDVNYNTNFSFTVPEGE Y LGDDRLVSSDSRHVGTFKAKDITGEAKFRLWPITRIGTF
SP107 nucleotide (SEQ ID NO:187)
GGACTCTCTCAAAGATGTGAAAGCAAATGCTAGCGACAGCAAGCCTGCACAGGACAAGAAGGATGCAAA ACAAGGAACGGAAGATAGTAAGGATTCAGATAAGATGACTGAAACAAACTCAGTTCCGGCAGGAGTGAT TGTGGTCAGTCTACTTGCCCTCCTAGGCGTGATTGCCTTCTGGCTGATTCGCCGTAAGAAAGAGTCAGA AATCCAGCAATTAAGCACGGAATTGATCAAGGTTCTAGGACAGCTAGATGCAGAAAAAGCGGATAAAAA AGTCCTTGCCAAAGCCCAAAACCTTCTCCAAGAAACCCTTGATTTCGTGAAAGAAGAAAATGGCTCAGC AGAGACAGAAACTAAACTAGTAGAGGAGCTTAAAGCAATCCTTGACAAAC CAAG
SP107 amino acid (SEQ ID NO: 188)
3SLKDVKANASDSKPAQDKKDAKQGTEDSKDSDKMTETNSVPAGVIWSLLA LGVIAFWLIRRKKESE IQQLSTE IKVLGQLDAEKADKKVLAKAQNLLQETLDFVKEENGSAETETKLVEELKAILDKLK
SP108 nucleotide (SEQ ID NO:189)
CAAGAAATCCTATCATCTCTTCCAGAAGCAAACAGAGACGAGGGGAATTCAGACTCAGTTGATTGAAGA ATCGCTTAGTCAGCAGACTATAATCCAGTCCTTCAATGCTCAAACAGAATTTATCCAAAGATTGCGTGA GGCTCATGACAACTACTCAGGCTATTCTCAGTCAGCCATCTTTTATTCTTCAACGGTCAATCCTTCGAC TCGCTTTGTAAATGCACTCATTTATGCCCTTTTAGCTGGAGTAGGAGCTTATCGTATCATGATGGGTTC AGCCTTGACCGTCGGTCGTTTAGTGACTTTTTTGAACTATGTTCAGCAATACACCAAGCCCTTTAACGA TATTTCTTCAGTGCTAGCTGAGTTGCAAAGTGCTCTGGCTTGCGTAGAGCGTATCTATGGAGTCTTAGA TAGCCCTGAAGTGGCTGAAACAGGTAAGGAAGTCTTGACGACCAGTGACCAAGTTAAGGGAGCTATTTC CTTTAAACATGTCTCTTTTGGCTACCATCCTGAAAAAATTTTGATTAAGGACTTGTCTATCGATATTCC AGCTGGTAGTAAGGTAGCCATCGTTGGTCCGACAGGTGCTGGAAAATCAACTCTTATCAATCTCCTTAT GCGTTTTTATCCCATTAGCTCGGGAGATATCTTGCTGGATGGGCAATCCATTTATGATTATACACGAGT ATCATTGAGACAGCAGTTTGGTATGGTGCTTCAAGAAACCTGGCTCACACAAGGGACCATTCATGATAA TATTGCCTTTGGCAATCCTGAAGCCAGTCGAGAGCAAGTAATTGCTGCTGCCAAAGCAGCTAATGCAGA CTTTTTCATCCAACAGTTGCCACAGGGATACGATACCAAGTTGGAAAATGCTGGAGAATCTCTCTCTGT CGGCCAAGCTCAGCTCTTGACCATAGCCCGAGTCTTTCTGGCTATTCCAAAGATTCTTATCTTAGACGA GGCAACTTCTTCCATTGATACACGGACAGAAGTGCTGGTACAGGATGCCTTTGCAAAACTCATGAAGGG CCGCACAAGTTTCATCATTGCTCACCGTTTGTCAACCATTCAGGATGCGGATTTAATTCTTGTCTTAGT Table 1 87
AGATGGTGATATTGTTGAATATGGTAACCATCAAGAACTCATGGATAGAAAGGGTAAGTATTACCAAAT
GCAAAAAGCTGCGGCTTTTAGTTCTGA
A
SP108 amino acid (SEQ ID NO:190)
KKSYHLFQKQTETRGIQTQLIΞESLSQQTIIQSFNAQTEFIQRLREAHDNYSGYSQSAIFYSSTVNPST RFVNA IYALLAGVGAYRIMMGSA TVGRLVTFLNYVQQYTKPFNDISSVLAE QSALACVERIYGVLD SPEVAETGKEVLTTSDQVKGAISFKHVΞFGYHPEKI IKDLSIDIPAGSKVAIVGPTGAGKSTLINLLM RFYPISSGDI LDGQSIYDYTRVSLRQQFG VLQET LTQGTIHDNIAFGNPEASRE VIAAAKAANAD FFIQQLPQGYDTK ENAGESLSVGQAQLLTIARVFLAIPKILI DEATSSIDTRTEV VQDAFAKLMKG RTSFIIAHRLSTIQDADLILVLVDGDIVEYGNHQELMDRKGKYYQMQKAAAFΞSE
SP109 nucleotide (SEQ ID NO: 191)
ACGAAATGCAGGGCAGACAGATGCCTCGCAAATTGAAAAGGCGGCAGTTAGCCAAGGAGGAAAAGCAGT GAAAAAAACAGAAATTAGTAAAGACGCAGACTTGCACGAAATTTATCTAGCTGGAGGTTGTTTCTGGGG AGTGGAGGAATATTTCTCACGTGTTCCCGGGGTGACGGATGCCGTTTCAGGCTATGCAAATGGTAGAGG AGAAACAACCAAGTACGAATTGATTAACCAAACAGGTCATGCAGAAACCGTCCATGTCACCTATGATGC CAAGCAAATTTCTCTCAAGGAAATCCTGCTTCACTATTTCCGCATTATCAATCCAACCAGCAAAAATAA ACAAGGAAATGATGTGGGGACCCAGTACCGTACTGGTGTTTATTACACAGATGACAAGGATTTGGAAGT GATTAACCAAGTCTTTGATGAGGTGGCTAAGAAATACGATCAACCTCTAGCAGTTGAAAAGGAAAACTT GAAGAATTTTGTGGTGGCTGAGGATTACCATCAAGACTATCTCAAGAAAAATCCAAATGGCTACTGCCA TATCAATGTTAATCAGGCGGCCTATCCTGTCATTGATGCCAGCAAATATCCAAAACCAAGTGATGAGGA ATTGAAAAAGACCCTGTCACCTGAGGAGTATGCAGTTACCCAGGAAAATCAAACAGAACGAGCTTTCTC AAACCGTTACTGGGATAAATTTGAATCCGGTATCTATGTGGATATAGCAACTGGGGAACCTCTCTTTTC ATCAAAAGACAAATTTGAGTCTGGTTGTGGCTGGCCTAGTTTTACCCAACCCATCAGTCCAGATGTTGT CACCTACAAGGAAGATAAGTCCTACAATATGACGCGTATGGAAGTGCGGAGCCGAGTAGGAGATTCTCA CCTTGGGCATGTCTTTACGGATGGTCCACAGGAC.2VAGGGCGGCTTACGTTACTGTATCAATAGCCTCTC TATCCGCTTTATTCCCAAAGACCAAATGGAAGAAAAAGGCTACGCTTATTTACTAGATTATGTTGAT
SP109 amino acid (SEQ ID NO: 192)
RNAGQTDASQIEKAAVSQGGKAV KTEISKDADLHEIY AGGCF GVEEYFSRVPGVTDAVSGYA GRG ETTKYELINQTGHAETVHVTYDAKQISLKEIL KYFRIINPTSKNKQGNDVGTQYRTGVYTDDKDLEV INQVFDEVAKKYDQPLAVEKENLKNFWAEDYHQDYLKKNPNGYCHINVNQAAYPVIDASKYPKPSDEΞ LKKTLSPEEYAVTQENQTERAFSNRY DKFESGIYVDIATGEPLFSSKDKFESGCG PSFTQPISPDW TYKEDKSYNMTRMEVRSRVGDSHLGhTVFTDGPQDKGGLRYCINS SIRFIPKDQMEEKGYAYL DYVD
SP110 nucleotide (SEQ ID NO: 193)
TGTATAGTTTTTAGCGCTTGTTCTTCTAATTCTGNTAAAAATGAAGAAAATACTTCTAAAGAGCATGCG CCTGATAAAATAGTTTTAGATCATGCTTTCGGTCAAACTATATTAGATAAAAAACCTGAAAGAGTTGCA ACTATTGCTTGGGGAAATCATGATGTAGCATTAGCTTTAGGAATAGTTCCTGTTGGATTTTCAAAAGCA AATTACGGTGTAAGTGCTGATAAAGGAGTTTTACCATGGACAGAAGAAAAAATCAAAGAACTAAATGGT AAAGCTAACCTATTTGACGATTTGGATGGACTTAACTTTGAAGCAATATCAAATTCTAAACCAGATGTT ATCTTAGCAGGTTATTCTGGTATAACTAAAGAAGATTATGACACTCTATCA
SP110 amino acid (SEQ ID NO:194)
CIVFSACSSNSXKNEENTSKEHAPDKIVLDHAFGQTI DKKPERVA IA GNHDVALALGIVPVGFSKA NYGVSADKGVLP TEEKIKELNGKAN FDDLDGLNFEAISNSKPDVILAGYSGITKEDYDTLS
SP111 nucleotide (SEQ ID NO: 195)
GTGTGTCGAGCATATTCTGAAGCAAACCTATCAAAATATAGAAATTATTTTAGTTGATGACGGTTCTAC GGATAATTCTGGGGAAATTTGTGATGCTTTTATGATGCAAGATAATCGTGTGCGAGTATTGCATCAAGA AAATAAGGGGGGGGCAGCACAAGCTAAAAATATGGGGATTAGTGTAGCTAAGGGAGAGTACATCACGAT TGTTGATTCAGATGATATCGTAAAAGAAAATATGATTGAAACTCTTTATCAGCAAGTCCAAGAAAAGGA TGCAGATGTTGTTATAGGGAATTACTATAATTATGACGAAAGTGACGGGAATTTTTATTTTTATGTAAC AGGGCAAGATT TTGCGTCGAAGAATTAGCTATACAAGAAATTATGAACCGTCAAGCAGGAGATTGGAA ATTCAATAGCTCGGCCTTTATATTGCCGACATTTAAGTTGATTAAAAAAGAATTATTCAATGAAGTTCA CTTTTCAAATGGTCGCCGCTTTGATGATGAAGCAACTATGCATCGCTTTTATCTTTTAGCCTCTAAAAT CGTCTTTATAAACGATAATCTCTATCTGTATAGAAGACGTTCAGGAAGCATCATGAGAACGGAATTTGA Table 1 88
TCTTTCCTGGGCAAGAGATATTGTTGAAGTGTTTTCTAAGAAAATATCGGATTGTGTCTTGGCTGGTTT GGATGTCTCCGTTCTGCGTATTCGATTTGTCAATCTTTTAAAAGATTATAAGCAAACTTTAGAATACCA TCAATTAACAGATACTGAGGAATATAAAGATATTTGTTTCAGATTAAAGTTGTTTTTTGATGCAGAACA AAGAAATGGTAAAAGT
SP111 amino acid (SEQ ID NO: 196)
O/EHILKQTYQNIEIILVDDGSTDNSGEICDAFMMQDNRVRVLHQENKGGAAQAKNMGISVAKGEYITI VDSDDIV ENMIETLYQQVQEKDADWIGNYYNYDESDGNFYFYVTGQDFCVEELAIQΞIMNRQAGDWK FNΞΞAFILPTFKLIKKE FNEVHFSNGRRFDDEATMHRFYLLASKIVFINDNLYLYRRRSGSIMRTEFD S ARDIVEVFSKKIΞDCVLAGLDVSVLRIRFVNLLKDYKQTLEYHQLTDTEΞYKDICFRLKLFFDAEQ RNGKS
SP0112 nucleotide (SEQ ID NO:197)
GTGTTTGGATAGCATTCAGAATCAGACGTATCAAAATTTTGAGTGTTTATTAATCAATGATGGCTCTCC AGATCATTCATCCAAAATATGTGAAGAATTTGTAGAGAAAGATTCTCGTTTCAAATATT TGAGAAAGC AAACGGCGGTCTTTCATCAGCTCGTAACCTAGGTATTGAATGTTCGGGGGGGGCGTACATTACTTTTGT AGACTCTGATGATTGGTTGGAACATGATGCTTTAGACCGATTATATGGTGCTTTGAAAAAGGAAAACGC AGATATTAGTATCGGGCGTTATAATTCTTATGATGAAACACGCTATGTGTATATGACTTATGTTACGGA TCCAGATGATTCTCTAGAAGTGATAGAAGGTAAAGCAATTATGGATAGGGAAGGTGTCGAAGAAGTCAG AAATGGGAACTGGACTGTAGCTGTCTTGAAGTTATTCAAGAGAGAGTTACTACAAGATTTACCATTTCC TATAGGAAAAATTGCAGAGGATACTTACTGGACATGGAAGGTACTTCTAAGAGCTTCGAGGATAGTCTA TTTGAATCGTTGTGTTTACTGGTACCGTGTTGGTTTATCTGATACTTTATCGAATACATGGAGTGAAAA GCGTATGTATGATGAAATTGGGGCTAGGGAAGAAAAGATAGCTATTTTAGCAAGTTCAGACTATGACTT GACCAATCATATTTTGATTTATAAAAATAGATTACAAAGAGTGATAGCAAAATTAGAAGAACAAAATAT GCAGTTCACAGAGATTTACAGAAGAATGATGGAAAAATTGTCTTTACTTCCG
SP0112 amino acid (SEQ ID NO:198)
CLDSIQNQTYQNFECLLINDGSPDHSSKICEΞFVEKDSRFKYFEKANGGL3SARNLGIΞCSGGAYITFV DSDDW EHDA DRLYGALKKENADISIGRYNSYDETRYVYMTYVTDPDDSLEVIEGKAI DREGVEEVR NGNWTVAV KLFKRELLQDLPFPIGKIAEDTY TWKV LRASRIVYLNRCVYWYRVG SDT SNT SEK RMYDEIGAREEKIAILASSDYDLTNHILIYKNRLQRVIAKLEEQ MQFTEIYRRMMEKLS LP
SP113 nucleotide (SEQ ID NO:199)
GTGCCTAGATAGTATTATTACTCAAACATATAAAAATATTGAGATTGTTGTCGTTAATGATGGTTCTAC GGATGCTTCAGGTGAAATTTGTAAAGAATTTTCAGAAATGGATCACCGAATTCTCTATATAGAACAAGA AAATGCTGGTCTTTCTGCCGCACGAAACACCGGTCTGAATAATATGTCCGGAAATTATGTGACCTTTGT GGACTCGGATGATTGGATTGAGCAAGATTATGTAGAAACTCTATATAAAAAAATAGTAGAGTATCAGGC TGATATTGCAGTTGGTAATTATTATTCTTTCAACGAAAGTGAAGGAATGTTCTACTTTCATATATTGGG AGACTCCTATTATGAGAAAGTATATGATAATGTTTCTATCTTTGAGAACTTGTATGAAACTCAAGAAAT GAAGAGTTTTGCTTTGATATCTGCTTGGGGTAAACTCTATAAGGCAAGATTGTTTGAGCAGTTGCGCTT TGACATAGGTAAATTAGGAGAAGATGGTTACCTCAATCAAAAGGTATATTTATTATCAGAAAAGGTAAT TTATTTAAATAAAAGTCTTTATGCTTATCGGATTAGAAAAGGTAGTTTATCAAGAGTTTGGACAGAAAA GTGGATGCACGCTTTAGTTGATGCTATGTCTGAACGTATTACGCTACTAGCTAATATGGGTTATCCTCT AGAGAAACACTTGGCAGTTTATCGTCAGATGTTGGAAGTCAGTCTCGCCAACGGTCAAGCTAGTGGTTT ATCTGACACAGCAACGTATAAAGAGTTTGAAATGAAACAAAGGCTTTTAAATCAGCTATCGAGACAAGA GGAAAGTGAAAAGAAAGCCATTGTCCTCGCAGCAAACTATGGCTATGTAGACCAAGTTTTAACGACAAT CAAGTCTATTTGTTATCATAATCGTTCGATTCGTTTTTATCTGATTCATAGCGATTTTCCAAATGAATG GATTAAGCAATTAAATAAGCGCTTAGAGAAGTTTGACTCAGAAATTATTAATTGTCGGGTAACTTCTGA GCAAATTTCATGTTATAAATCGGATATTAGTTACACAGTCTTTTTACGCTATTTCATAGCTGATTTCGT GCAAGAAGACAAGGCCCTCTACTTGGACTGTGATCTAGTTGTAACGAAAAATCTGGATGACTTGTTTGC TACAGACTTACAAGATTATCCTTTGGCTGCTGTTAGAGATTTTGGGGGCAGAGCTTATTTTGGTCAAGA AATCTTTAATGCCGGTGTTCTCTTGGTAAACAATGCTTTTTGGAAAAAAGAGAATATGACCCAAAAATT AATTGATGTAACCAATGAATGGCATGATAAGGTGGATCAGGCAGATCAGAGCATCTTGAATATGCTTTT TGAACATAAATGGTTGGAATTGGACTTTGATTATAATCATATTGTCATTCATAAACAGTTTGCTGATTA TCAATTGCCTGAGGGTCAGGATTATCCTGCTATTATTCACTATCTTTCTCATCGGAAACCGTGGAAAGA TTTGGCGGCCCAAACCTATCGTGAAGTTTGGTGGTACTATCATGGGCTTGAATGGACAGAATTGGGACA AAACCATCATTTACATCCATTACAAAGATCTCACATCTATCCAATAAAGGAACCTTTCACTTGTCTAAT CTATACTGCCTCAGACCATATTGAACAAATTGAGACATTGGTTCAATCCTTGCCTGATATTCAGTTTAA Table 1 89
GATAGCAGCTAGAGTAATAGTTAGTGATCGATTGGCTCAGATGACAATTTATCCAAACGTGACTATATT TAACGGAATTCACTATTTGGTAGATGTCGATAATGAATTGGTAGAAACCAGTCAAGTACTTTTAGATAT TAATCATGGCGAAAAGACAGAAGAAATTCTCGATCAATTTGCTAATCTTGGCAAGCCTATCTTATCCTT TGAAAATACTAAAACCTATGAAGTAGGTCAGGAGGCATATGCTGTTGACCAAGTTCAAGCAATGATTGA AAAATTGAGAGAAATAAGCAAA
SP113 amino acid (SEQ ID NO:200)
CLDSIITQTYKNIEIVWNDGSTDASGEICKEFSEMDHRILYIEQENAG SAARNTGL NMSGNYVTFV DSDDWIEQDYVETLYKKIVEYQADIAVG YYSFNESEG FYFHI GDSYYEKVYDNVSIFENLYETQEM KSFALISA GKLYKARLFEQLRFDIGKLGEDGYLNQKVYLLSEKVIYLNKSLYAYRIRKGSLSRVWTEK MHALVDAMSERITLLANMGYPLΞKH AVYRQM ΞVSLANGQASGLSDTATYKEFEMKQR LNQLSRQE ΞSEKKAIVLAANYGYVDQVLTTIKSICYHNRSIRFYLIHSDFPNE IKQLNKRLEKFDSΞIINCRVTSE QISCYKSDISYTVF RYFIADFVQEDKALYLDCDLWTKNLDDLFATD QDYPLAAVRDFGGRAYFGQE IFNAGVLLVNNAFWKKENMTQK IDVTNEWHDKVDQADQSILNMLFEHKWLΞLDFDYNKIVIHKQFADY QLPEGQDYPAIIKYLSHRKPWKDLAAQTYREVW YYHGLΞ TELGQNHHLHPLQRSHIYPIKEPFTCLI YTASDHIEQIET VQS PDIQFKIAARVIVSDRAQMTIYPNVTIFNGIHYLVDVDNELVΞTSQVLLDI NHGEKTEEILDQFANLGKPILSFΞNTKTYΞVGQEAYAVDQVQAMIΞKLREIΞK
SP114 nucleotide (SEQ ID NO:201)
CATTCAGAAGCAGACCTATCAAAATCTGGAAATTATTCTTGTTGATGATGGTGCAACAGATGAAAGTGG TCGCTTGTGTGATTCAATCGCTGAACAAGATGACAGGGTGTCAGTGCTTCATAAAAAGAACGAAGGATT GTCGCAAGCACGAAATGATGGGATGAAGCAGGCTCACGGGGATTATCTGATTTTTATTGACTCAGATGA TTATATCCATCCAGAAATGATTCAGAGCTTATATGAGCAATTAGTTCAAGAAGATGCGGATGTTTCGAG CTGTGGTGTCATGAATGTCTATGCTAATGATGAAAGCCCACAGTCAGCCAATCAGGATGACTATTTTGT CTGTGATTCTCAAACATTTCTAAAGGAATACCTCATAGGTGAAAAAATACCTGGGACGA 'TTGCAATAA GCTAATCAAGAGACAGATTGCAACTGCCCTATCCTTTCCTAAGGGGTTGATTTACGAAGATGCCTATTA CCATTTTGATTTAATCAAGTTGGCCAAGAAGTATGTGGTTAATACTAAACCCTATTATTACTATTTCCA TAGAGGGGATAGTATTACGACCAAACCCTATGCAGAGAAGGATTTAGCCTATATTGATATCTACCAAAA GTTTTATAATGAAGTTGTGAAAAACTATCCTGACTTGAAAGAGGTCGCTTTTTTCAGAT GGCCTATGC CCACTTCTTTATTCTGGATAAGATGTTGCTAGATGATCAGTATAAACAGTTTGAAGCCTATTCTCAGAT TCATCGTTTTTTAAAAGGCCATGCCTTTGCTATTTCTAGGAATCCAATTTTCCGTAAGGGGAGAAGAAT TAGTGCTTTGGCCCTATTCATAAATATTTCCTTATATCGATTCTTATTACTGAAAAATATTGAAAAATC TAAAAAATTACAT
SP114 amino acid (SEQ ID NO:202)
IQKQTYQNLΞIILVϋDGATDESGRLCDSIAEQDDRVSVLHKKNEGLSQARNDGMKQAHGDY IFIDSDD YIHPE IQSLYEQLVQEDADVSSCGVMNVYANDESPQSANQDDYFVCDSQTFLKEYLIGEKIPGTICNK IKRQIATALSFPKGLIYEDAYYHFDLIKLAKKYWNTKPYYYYFHRGDSITTKPYAEK3LAYIDIYQK FYNEWK YPDLKΞVAFFRLAYAHFFILDKMLLDDQYKQFEAYSQIHRFLKGHAFAISRNPIFRKGRRI SALALFINI5 YRFLLLKNIΞKSKKLH
SP115 nucleotide (SEQ ID NO:203)
TAAGGCTGATAATCGTGTTCAAATGAGAACGACGATTAATAATGAATCGCCATTGTTGCTTTCTCCGTT GTATGGCAATGATAATGGTAACGGATTATGGTGGGGGAACACATTGAAGGGAGCATGGGAAGCTATTCC TGAAGATGTAAAGCCATATGCAGCGATTGAACTTCATCCTGCAAAAGTCTGTAAACCAACAAGTTGTAT TCCACGAGATACGAAAGAATTGAGAGAATGGTATGTCAAGATGTTGGAGGAAGCTCAAAGTCTAAACAT TCCAGTTTTCTTGGTTATTATGTCGGCTGGAGAGCGTAATACAGTTCCTCCAGAGTGGTTAGATGAACA ATTCCAAAAGTATAGTGTGTTAAAAGGTGTTTTAAATATTGAGAATTATTGGATTTACAATAACCAGTT AGCTCCGCATAGTGCTAAATATTTGGAAGTTTGTGCCAAATATGGAGCGCATTTTATCTGGCATGATCA TGAAAAATGGTTCTGGGAAACTATTATGAATGATCCGACATTCTTTGAAGCGAGTCAAAAATATCATAA AAATTTGGTGTTGGCAACTAAAAATACGCCAATAAGAGATGATGCGGGTACAGATTCTATCGTTAGTGG ATTTTGGTTGAGTGGCTTATGTGATAACTGGGGCTCATCAACAGATACATGGAAATGGTGGGAAAAACA TTATACAAACACATTTGAAACTGGAAGAGCTAGGGATATGAGATCCTATGCATCGGAACCAGAATCAAT GATTGCTATGGAAATGATGAATGTATATACTGGGGGAGGCACAGTTTATAATTTCGAATGTGCCGCGTA TACATTTATGACAAATGATGTACCAACTCCAGCATTTACTAAAGGTATTATTCCTTTCTTTAGACATGC TATACAAAATCCAGCTCCAAGTAAGGAAGAAGTTGTAAATAGAACAAAAGCTGTATTTTGGAATGGAGA AGGTAGGATTAG TCATTAAACGGATTTTATCAAGGACTTTATTCGAATGATGAAACAATGCCTTTATA TAATAATGGGAGATATCATATTCTTCCTGTAATACATGAGAAAATTGATAAGGAAAAGATTTCATCTAT Table 1 90
ATTCCCTAATGCAAAAATTTTGACTAAAAATAGTGAGGAATTGTCTAGTAAAGTCAACTATTTAAACTC GCTTTATCCAAAACTTTATGAAGGAGATGGGTATGCTCAGCGTGTAGGTAATTCCTGGTATATTTATAA TAGTAATGCTAATATCAATAAAAATCAGCAAGTAATGTTGCCTATGTATACTAATAATACAAAGTCGTT ATCGTTAGATTTGACGCCACATACTTACGCTGTTGTTAAAGAAAATCCAAATAATTTACATATTTTATT GAATAATTACAGGACAGATAAGACAGCTATGTGGGCATTATCAGGAAATTTTGATGCATCAAAAAGTTG GAAGAAAGAAGAATTAGAGTTAGCGAACTGGATAAGCAAAAATTATTCCATCAATCCTGTAGATAATGA CTTTAGGACAACAACACTTACATTAAAAGGGCATACTGGTCATAAACCTCAGATAAATATAAGTGGCGA TAAAAATCATTATACTTATACAGAAAATTGGGATGAGAATACCCATGTTTATACCATTACGGTTAATCA TAATGGAATGGTAGAGATGTCTATAAATACTGAGGGGACAGGTCCAGTCTCTTTCCCAACACCAGATAA ATTTAATGATGGTAATTTGAATATAGCATATGCAAAACCAACAACACAAAGTTCTGTAGATTACAATGG AGACCCTAATAGAGCTGTGGATGGTAACAGAAATGGTAATTTTAACTCTGGTTCGGTAACACACACTAG GGCAGATAATCCCTCTTGGTGGGAAGTCGATTTGAAAAAAATGGATAAAGTTGGGCTTGTTAAAATTTA TAATCGCACAGATGCTGAGACTCAACGTCTATCTAATTTT
SP115 amino acid (SEQ ID NO:204)
KADNRVQMRTTINNESPLLLSPLYGNDNGNGLWWGNTLKGAWEAIPEDVKPYAAIELHPAKVCKPTSCI PRDTKE REWYVKMLEEAQSLNIPVF VIMSAGERNTVPPEWLDEQFQKYSVLKGVLNIΞNY IYNNQL APHSAKYLEVCAKYGAHFIWHDHEK F ETIMNDPTFFEASQKYKKNLVLATKNTPIRDDAGTDSIVSG FWLSGLCDN GSSTDT KWWEKHYTNTFΞTGRARDMRΞYASEPΞSMIAME ]yπWYTGGGTΛ/YNFECAAY TFMTNDVPTPAFTKGIIPFFRHAIQNPAPSKΞEWNRTKAVFWNGEGRISSLNGFYQGLYΞNDETMPLY INGRYHI PVIKΞKIDKEKISSIF?NAKI TK^SEELSΞKVKr/LNS YPK YΞGDGYAQRVGNSW'IYN SNANINKNQQVMLP YTNNTKSLSLDLTPHTYAVVKENPNNLKI NNYRTDKTAMWA SGNFDASKS KKEE ELANWISKNYSINPVDNDFRTTT KGHTGHKPQINISGDK^HYTYTΞN DENTHVYTITVNH NG VE SINTEGTGPVSFPTPDKFNDGNLNIAYAKPTTQS3VDYNGDPNRAVDGNRNGNFNSGSVTHTR ADNPS WEVDLKK DKVGLVKIYNRTDAETQRLSNF
SP117 nucleotide (SEQ ID NO:205)
CTGTGGCAATCAGTCAGCTGCTTCCAAACAGTCAGCTTCAGGAACGATTGAGGTGATTTCACGAGAAAA TGGCTCTGGGACACGGGGTGCCTTCACAGAAATCACAGGGATTCTCAAAAAAGACGGTGATAAAAAAAT TGACAACACTGCCAAAACAGCTGTGATTCAAAATAGTACAGAAGGTGTTCTCTCAGCAGTTCAAGGGAA TGCTAATGCTATCGGCTACATCTCCTTGGGATCTTTAACGAAATCTGTCAAGGCTTTAGAGATTGATGG TGTCAAGGCTAGTCGAGACACAGTTTTAGATGGTGAATACCCTCTTCAACGTCCCTTCAACATTGTTTG GTCTTCTAATCTTTCCAAGCTAGGTCAAGATTTTATCAGCTTTATCCACTCCAAACAAGGTCAACAAGT GGTCACAGATAATAAATTTATTGAAGCTAAAACCGAAACCACGGAATATACAAGCCAACACTTATCAGG CAAGTTGTCTGTTGTAGGTTCCACTTCAGTATCTTCTTTAATGGAAAAATTAGCAGAAGCTTATAAAAA AGAAAATCCAGAAGTTACGATTGATAT ACCTCTAATGGGTCTTCAGCAGGTATTACCGCTGTTAAGGA GAAAACCGCTGATATTGGTATGGTTTCTAGGGAΛTTAACTCCTGAAGAAGGTAAGAGTCTCACCCATGA TGCTATTGCTTTAGACGGTATTGCTGTTGTGGTCAATAATGACAATAAGGCAAGCCAAGTCAGTATGGC TGAACTTGCAGACGTTTTTAGTGGCAAATTAACCACCTGGGACAAGATTAAA
SP117 amino acid (SEQ ID NO: 206)
CGNQSAASKQSASGTIEVISRENGSGTRGAFTEITGILKKDGDKKIDNTAKTAVIQNSTEGVLSAVQGN ANAIGYISLGSLTKSVKALEIDGVKASRDTVLDGEYPLQRPFNIVWSSN SKLGQDFISFIHSKQGQQV VTDNKFIEAKTETTΞYTSQHLSGKLSWGSTSVSSLMEKLAEAYKKENPEVTIDITSNGSSAGITAVKE KTADIGMVSRELTPEEGKSLTHDAIALDGIAVVVNNDNKASQVSMAELADVFSGKLTTWDKIK
SP118 nucleotide (SEQ ID NO:207)
TTGTCAACAACAACATGCTACTTCTGAGGGGACGAATCAAAGGCAAAGCAGTTCAGCGAAAGTTCCATG GAAAGCTTCATACACCAACCTAAACAACCAGGTAAGTACAGAAGAGGTCAAATCTCTCTTATCAGCTCA CTTGGATCCAAATAGTGTTGATGCATTTTTTAATCTCGTTAATGACTATAATACCATTGTCGGCTCAAC TGGCTTATCAGGAGATTTCACTTCCTTTACTCACACCGAATACGATGTTGAGAAAATCAGTCATCTCTG GAATCAAAAGAAGGGCGATTTTGTTGGGACCAACTGCCGTATCAATAGTTATTGTCTTTTGAAAAATTC AGTCACCATTCCAAAGCTTGAAAAGAATGACCAGTTGCTTTTCCTAGATAATGATGCGATTGATAAAGG AAAGGTCTTTGATTCACAAGATAAGGAAGAGT'TTGATATTCTATT TCGAGAGTTCCAACTGAGTCAAC TACAGATGTCAAGGTTCACGCTGAAAAGATGGAAGCATTCTTCTCACAATTTCAATTCAATGAAAAAGC TCGAATGCTGTCTGTAGTCTTGCACGACAATTTGGATGGCGAGTATCTGTTTGTAGGCCACGTTGGGGT CTTAGTACCTGCTGATGACGGTTTCTTATTTGTAGAGAAATTGACTTTCGAAGAGCCCTACCAAGCGAT Table 1 91
TAAATTTGCTAGTAAGGAAGATTGCTACAAGTATTTGGGCACCAAGTATGCGGATTATACAGGCGAGGG ACTGGCTAAGCCTTTTATCATGGATAATGATAAGTGGGTTAAACTT
SP118 amino acid (SEQ ID NO:208)
CQQQHATSEGTNQRQSSSAKVP KASYTNLNNQVSTEEVKS LSAHLDPNSVDAFFNLVNDYNTIVGST G SGDFTSFTHTEYDVEKISHLWNQKKGDFVGTNCRINSYC LKNΞVTIPKLEKNDQLLF DNDAIDKG KVFDSQDKEEFDI FSRVPTESTTDVTVHAEKMEAFFSQFQFNEKARMLSVVLHDNLDGEYLFVGHVGV LVPADDGFLFVEK TFEEPYQAIKFASKEDCYKYLGTKYADYTGEG AKPFIMDNDK VKL
SP119 nucleotide (SEQ ID NO: 209)
TTGTTCAGGCAAGTCCGTGACTAGTGAACACCAAACGAAAGATGAAATGAAGACGGAGCAGACAGCTAG TAAAACAAGCGCAGCTAAAGGGAAAGAGGTGGCTGATTTTGAATTGATGGGAGTAGATGGCAAGACCTA CCGTTTATCTGATTACAAGGGCAAGAAAGTCTATCTCAAATTCTGGGCTTCTTGGTGTTCCATCTGTCT GGCTAGTCTTCCAGATACGGATGAGATTGCTAAAGAAGCTGGTGATGACTATGTGGTCTTGACAGTAGT GTCACCAGGACATAAGGGAGAGCAATCTGAAGCGGACTTTAAGAATTGGTATAAGGGATTGGATTATAA AAATCTCCCAGTCCTAGTTGACCCATCAGGCAAACTTTTGGAAACTTATGGTGTCCGTTCTTACCCAAC CCAAGCCTTTATAGACAAAGAAGGCAAGCTGGTCAAAACACATCCAGGATTCATGGAAAAAGATGCAAT TTTGCAAACTTTGAAGGAATTAGCC
SP119 amino acid (SEQ ID NO:210)
CSGKSVTSEHQTKDEMKTEQTASKTSAAKGKEVADFELMGVDGKTYRLSDYKGKKVYLKF ASWCSIC ASLPDTDEIAKEAGDDYVVLTVVSPGHKGEQSEADFKNWYKGLDYKNLPVLVDPSGKLLETYGVRSYPT QAFIDKEGKLVKTHPGFMEKDAILQTLKΞLA
SP120 nucleotide (SEQ ID NO:211)
CTCGCAAATTGAAAAGGCGGCAGTTAGCCAAGGAGGAAAAGCAGTGAAAAAAACAGAAATTAGTAAAGA CGCAGACTTGCACGAAATTTATCTAGCTGGAGGTTGTTTCTGGGGAGTGGAGGAATATTTCTCACGTGT TCCCGGGGTGACGGATGCCGTTTCAGGCTATGCAAATGGTAGAGGAGAAACAACCAAGTACGAATTGAT TAACCAAACAGGTCATGCAGAAACCGTCCATGTCACCTATGATGCCAAGCAAATTTCTCTCAAGGAAAT CCTGCTTCACTATTTCCGCATTATCAATCCAACCAGCAAAAATAAACAAGGAAATGATGTGGGGACCCA GTACCGTACTGGTGTTTATTACACAGATGACAAGGATTTGGAAGTGATTAACCAAGTCTTTGATGAGGT GGCTAAGAAATACGATCAACCTCTAGCAGTTGAAAAGGAAAACTTGAAGAATTTTGTGGTGGCTGAGGA TTACCATCAAGACTATCTCAAGAAAAATCCAAATGGCTACTGCCATATCAATGTTAATCAGGCGGCCTA TCCTGTCATTGATGCCAGCAAATATCCAAAACCAAGTGATGAGGAATTGAAAAAGACCCTGTCACCTGA GGAGTATGCAGTTACCCAGGAAAATCAAACAGAACGAGCTTTCTCAAACCGTTACTGGGATAAATTTGA ATCCGGTATCTATGTGGATATAGCAACTGGGGAACCTCTCTTTTCATCAAAAGACAAATTTGAGTCTGG TTGTGGCTGGCCTAGTTTTACCCAACCCATCAGTCCAGATGTTGTCACCTACAAGGAAGATAAGTCCTA CAATATGACGCGTATGGAAGTGCGGAGCCGAGTAGGAGATTCTCACCTTGGGCATGTCTTTACGGATGG TCCACAGGACAAGGGCGGCTTACGTTACTGTATCAATAGCCTCTCTATCCGCTTTATTCCCAAAGACCA AATGGAAGAAAAAGGTACGCTTATTTAC
SP120 amino acid (SEQ ID NO:212)
SQIEKAAVΞQGGKAVKKTEISKDADLKEIYLAGGCFWGVEΞYFSRVPGVTDAVSGYANGRGETTKYELI NQTGHAETVHVTYDAKQISLKEILLHYFRIINPTSKNKQGNDVGTQYRTGVYYTDDKDLEVINQVFDEV AKKYDQP AVEKENLKNFWAEDYHQDYLKKNPNGYCHI λNQAAYPVIDAΞKYPKPSDEELKKTLSPE EYAVTQENQTERAFSNRYWDKFESGIYVDIATGEPLFSSKDKFESGCGWPSFTQPISPDWTYKEDKSY NMTRMEVRSRVGDSHLGHVFTDGPQDKGGLRYCINSLSIRFIPKDQ EEKGTLIY
SP121 nucleotide (SEQ ID NO:213)
TTGTCAGTCAGGTTCTAATGGTTCTCAGTCTGCTGTGGATGCTATCAAACAAAAAGGGAAATTAGTTGT GGCAACCAGTCCTGACTATGCACCCTTTGAATTTCAATCATTGGTTGATGGAAAGAACCAGGTAGTCGG TGCAGACATCGACATGGCTCAGGCTATCGCTGATGAACTTGGGGTTAAGTTGGAAATCTCAAGCATGAG TTTTGACAATGTTTTGACCAGTCTTCAAACTGGTAAGGCTGACCTAGCAGTTGCAGGAATTAGTGCTAC TGACGAGAGAAAAGAAGTCTTTGATTTTTCAATCCCATACTATGAAAACAAGATTAGTTTCTTGGTTCG TAAGGCTGATGTGGAAAAATACAAGGATTTAACTAGCCTAGAAAGTGCTAATATTGCAGCCCAAAAAGG GACTGTTCCAGAATCAATGGTCAAGGAACAATTGCCAAAAGTTCAATTAACTTCCCTAACTAATATGGG TGAAGCAGTCAATGAATTGCAGGCTGGAAAAATAGATGCTGTTCATATGGATGAGCCTGTTGCACTTAG Table 1 92
TTATGCTGCTAAAAACGCTGGCTTAGCTGTCGCAACTGTCAGCTTGAAGATGAAGGACGGCGACGCCAA TGCC
SP121 amino acid (SEQ ID NO: 214)
CQSGSNGSQSAVDAIKQKGK WATSPDYAPFEFQSLVϋGKNQWGADIDMAQAIADELGVKLEISS S FDNVLTSLQTGKADLAVAGISATDERKEVFDFSIPYYENKISFLVRKADVEKYKDLTSLESANIAAQKG TVPEST^KEQLPKVQLTSLTNMGFJVNELQAGKIDAVHMDEPVALSYAAKNAGI_AVATVSLKMKDGDAN A
SP122 nucleotide (SEQ ID NO:215)
GGAAACTTCACAGGATTTTAAAGAGAAGAAAACAGCAGTCATTAAGGAAAAAGAAGTTGTTAGTAAAAA TCCTGTGATAGACAATAACACTAGCAATGAAGAAGCAAAAATCAAAGAAGAAAATTCCAATAAATCCCA AGGAGATTATACGGACTCATTTGTGAATAAAAACACAGAAAATCCCAAAAAAGAAGATAAAGTTGTCTA TATTGCTGAATTTAAAGATAAAGAATCTGGAGAAAAAGCAATCAAGGAACTATCCAGTCTTAAGAATAC AAAAGTTTTATATACTTATGATAGAATTTTTAACGGTAGTGCCATAGAAACAACTCCAGATAACTTGGA CAAAATTAAACAAATAGAAGGTATTTCATCGGTTGAAAGGGCACAAAAAGTCCAACCCATGATGAATCA TGCCAGAAAGGAAATTGGAGTTGAGGAAGCTATTGATTACCTAAAGTCTATCAATGCTCCGTTTGGGAA AAATTTTGATGGTAGAGGTATGGTCATTTCAAATATCGATACTGGAACAGATTATAGACATAAGGCTAT GAGAATCGATGATGATGCCAAAGCCTCAATGAGATTTAAAAAAGAAGACTTAAAAGGCACTGATAAAAA TTATTGGTTGAGTGATAAAATCCCTCATGCGTTCAATTATTATAATGGTGGCAAAATCACTGTAGAAAA ATATGATGATGGAAGGGATTATTTTGACCCACATGGGATGCATATTGCAGGGATTCTTGCTGGAAATGA TACTGAACAAGACATCAAAAACTTTAACGGCATAGATGGAATTGCACCTAATGCACAAATTTTCTCTTA CAAAATGTATTCTGACGCAGGATCTGGGTTTGCGGGTGATGAAACAATGTTTCATGCTATTGAAGATTC TATCAAACACAACGTTGATGTTGTTTCGGTATCATCTGGTTTTACAGGAACAGGTCTTGTAGGTGAGAA ATATTGGCAAGCTATTCGGGCATTAAGAAAAGCAGGCATTCCAATGGTTGTCGCTACGGGTAACTATGC GACTTCTGCTTCAAGTTCTTCATGGGATTTAGTAGCAAATAATCATCTGAAAATGACCGACACTGGAAA TGTAACACGAACTGCAGCACATGAAGATGCGATAGCGGTCGCTTCTGCTAAAAATCAAACAGTTGAGTT TGATAAAGTTAACATAGGTGGAGAAAGTTTTAAATACAGAAATATAGGGGCCTTTTTCGATAAGAGTAA AATCACAACAAATGAAGATGGAACAAAAGCTCCTAGTAAATTAAAATTTGTATATATAGGCAAGGGGCA AGACCAAGATTTGATAGGTTTGGATCTTAGGGGCAAAATTGCAGTAATGGATAGAATTTATACAAAGGA TTTAAAAAATGCTTTTAAAAAAGCTATGGATAAGGGTGCACGCGCCATTATGGTTGTAAATACTGTAAA TTACTACAATAGAGATAATTGGACAGAGCTTCCAGCTATGGGATATGAAGCGGATGAAGGTACTAAAAG TCAAGTGTTTTCAATTTCAGGAGATGATGGTGTAAAGCTATGGAACATGATTAATCCTGATAAAAAAAC TGAAGTCAAAAGAAATAATAAAGAAGATTTTAAAGATAAATTGGAGCAATACTATCCAATTGATATGGA AAGTTTTAATTCCAACAAACCGAATGTAGGTGACGAAAAAGAGATTGACTTTAAGTTTGCACCTGACAC AGACAAAGAACTCTATAAAGAAGATATCATCGTTCCAGCAGGATCTACATCTTGGGGGCCAAGAATAGA TTTACTTTTAAAACCCGATGTTTCAGCACCTGGTAAAAATATTAAATCCACGCTTAATGTTATTAATGG CAAATCAACTTATGGCTATATGTCAGGAACTAGTATGGCGACTCCAATCGTGGCAGCTTCTACTGTTTT GATTAGACCGAAATTAAAGGAAATGCTTGAAAGACCTGTATTGAAAAATCTTAAGGGAGATGACAAAAT AGATCTTACAAGTCTTACAAAAATTGCCCTACAAAATACTGCGCGACCTATGATGGATGCAACTTCTTG GAAAGAAAAAAGTCAATACTTTGCATCACCTAGACAACAGGGAGCAGGCCTAATTAATGTGGCCAATGC TTTGAGAAATGAAGTTGTAGCAACTTTCAAAAACACTGATTCTAAAGGTTTGGTAAACTCATATGGTTC CATTTCTCTTAAAGAAATAAAAGGTGATAAAAAATACTTTACAATCAAGCTTCACAATACATCAAACAG ACCTTTGACTTTTAAAGTTTCAGCATCAGCGATAACTACAGATTCTCTAACTGACAGATTAAAACTTGA TGAAACATATAAAGATGAAAAATCTCCAGATGGTAAGCAAATTGTTCCAGAAATTCACCCAGAAAAAGT CAAAGGAGCAAATATCACATTTGAGCATGATACTTTCACTATAGGCGCAAATTCTAGCTTTGATTTGAA TGCGGTTATAAATGTTGGAGAGGCCAAAAACAAAAATAAATTTGTAGAATCATTTATTCATTTTGAGTC AGTGGAAGCGATGGAAGCTCTAAACTCCAGCGGGAAGAAAATAAACTTCCAACCTTCTTTGTCGATGCC TCTAATGGGATTTGCTGGGAATTGGAACCACGAACCAATCCTTGATAAATGGGCTTGGGAAGAAGGGTC AAGATCAAAAACACTGGGAGGTTATGATGATGATGGTAAACCGAAAATTCCAGGAACCTTAAATAAGGG AATTGGTGGAGAACATGGTATAGATAAATTTAATCCAGCAGGAGTTATACAAAATAGAAAAGATAAAAA TACAACATCCCTGGATCAAAATCCAGAATTATTTGCTTTCAATAACGAAGGGATCAACGCTCCATCATC AΛGTGGTTCTAAGATTGCTAACATTTATCCTTTAGATTCAAATGGAAATCCTCAAGATGCTCAACTTGA AAGAGGATTAACACCTTCTCCACTTGTATTAAGAAGTGCAGAAGAAGGATTGATT
SP122 amino acid (SEQ ID NO:216)
ETSQDFKEKKTAVIKEKEVYSKNPVIDNNTSNEEAKIKEENSNKSQGDYTDSFλMKNTENPKKEDKVVY IAEFKDKESGEKAIKELSSLKNTKVLYTYDRIFNGΞAIETTPDN DKIKQIEGISSVERAQKVQPMMNH Table 1 93
ARKEIGVEEAIDY KSINAPFGK FDGRGMVISNIDTGTDYRHKAMRIDDDAKASMRFKKEDLKGTDKN YWLSDKIPHAFNYYNGGKITVEKYDDGRDYFDPHGMHIAGILAGNDTEQDIKNFNGIDGIAPNAQIFSY KMYSDAGΞGFAGDETMFHAIEDSIKHNVDVVSVSSGFTGTGLVGEKY QAIRALRKAGIPMVVATGNYA TSASSSS DLVAl^HLKMTDTGNVTRTAAHEDAIAVASAKNQTVEFDKVNIGGESFKYRNIGAFFDKSK ITTNEDGTK.APSKLKFVYIGKGQDQD IG DLRGKIAW1DRIYTKDLKNAFKKAMDKGARAIMVVNTVN YYNRDNWTE PAMGYEADEGTKSQVFSISGDDGVKLWNMINPDKKTEVKRNNKEDFKDK EQYYPIDME SFNSNKP VGDEKEIDFKFAPDTDKELYKEDIIVPAGSTSWGPRIDLLLKPDVSAPGKNIKSTL VING KSTYGYMSGTSMATPIVAASTVLIRPKLKEMLERPVLKNLKGDDKIDLTSLTKIALQNTARPMMDATS KEKSQYFASPRQQGAGLINVANALRNEWATFKNTDSKGLV SYGSISLKEIKGDKKYFTIK HNTSNR PLTFKVSASAITTDSLTDRLKLDETYKDEKSPDGKQIVPEIHPEKVKGA ITFEHDTFTIGANSSFDLN AVINVGEAKNKNKFVESFIHFESVEAMEALNSSGKKINFQPS SMPLMGFAGNWNHEPILDKWA EEGS RSKTLGGYDDDGKPKIPGTLNKGIGGEHGIDKFNPAGVIQNRKDKNTTSLDQNPE FAFNNEGINAPSS SGSKIANIYPLDSNGNPQDAQLERGLTPSPLVLRSAEEGLI
SP123 nucleotide (SEQ ID NO: 217)
TGTGGTCGAAGTTGAGACTCCTCAATCAATAACAAATCAGGAGCAAGCTAGGACAGAAAACCAAGTAGT AGAGACAGAGGAAGCTCCAAAAGAAGAAGCACCTAAAACAGAAGAAAGTCCAAAGGAAGAACCAAAATC GGAGGTAAAACCTACTGACGACACCCTTCCTAAAGTAGAAGAGGGGAAAGAAGATTCAGCAGAACCAGC TCCAGTTGAAGAAGTAGGTGGAGAAGTTGAGTCAAAACCAGAGGAAAAAGTAGCAGTTAAGCCAGAAAG TCAACCATCAGACAAACCAGCTGAGGAATCAAAAGTTGAACAAGCAGGTGAACCAGTCGCGCCAAGAGA AGACGAAAAGGCACCAGTCGAGCCAGAAAAGCAACCAGAAGCTCCTGAAGAAGAGAAGGCTGTAGAGGA AACACCGAAACAAGAAGAGTCAACTCCAGATACCAAGGCTGAAGAAACTGTAGAACCAAAAGAGGAGAC TGTTAATCAATCTATTGAACAACCAAAAGTTGAAACGCCTGCTGTAGAAAAACAAACAGAACCAACAGA GGAACCAAAAGTTGAACAAGCAGGTGAACCAGTCGCGCCAAGAGAAGACGAACAGGCACCAACGGCACC AGTTGAGCCAGAAAAGCAACCAGAAGTTCCTGAAGAAGAGAAGGCTGTAGAGGAAACACCGAAACCAGA AGATAAAATAAAGGGTATTGGTACTAAAGAACCAGTTGATAAAAGTGAGTTAAATAATCAAATTGATAA AGCTAGTTCAGTTTCTCCTACTGATTATTCTACAGCAAGTTACAATGCTCTTGGACCTGTTTTAGAAAC TGCAAAAGGTGTCTATGCTTCAGAGCCTGTAAAACAGCCTGAGGTAAATAGCGAGACAAATAAACTTAA AACGGCTATTGACGCTCTAAACGTTGATAAAACTGAATTAAACAATACGATTGCAGATGCAAAAACAAA GGTAAAAGAACATTACAGTGATAGAAGTTGGCAAAACCTCCAAACTGAAGTTACAAAGGCTGAAAAAGT TGCAGCTAATACAGATGCTAAACAAAGTGAAGTTAACGAAGCTGTTGAAAAATTAACTGCAACTATTGA AAAATTGGTTGAATTATCTGAAAAGCCAATATTAACATTGACTAGTACCGATAAGAAAATATTGGAACG TGAAGCTGTTGCTAAGTATACTCTAGAAAATCAAAACAAAACAAAAATCAAATCAATCACAGCTGAATT GAAAAAAGGAGAAGAAGTTATTAATACTGTAGTCCTTACAGATGACAAGGTAACAACAGAAACTATAAG CGCTGCATTTAAGAACCTAGAGTACTACAAAGAATACACCCTATCTACAACTATGATTTACGACAGAGG TAACGGTGAAGAAACTGAAACTCTAGAAAATCAAAATATTCAATTAGATCTTAAAAAAGTTGAGCTTAA AAATATTAAACGTACAGATTTAATCAAATACGAAAATGGAAAAGAAACTAATGAATCACTGATAACAAC TATTCCTGATGATAAGAGCAATTATTATTTAAAAATAACTTCAAATAATCAGAAAACTACATTACTAGC TGTTAAAAATATAGAAGAAACTACGGTTAACGGAACACCTGTATATAAAGTTACAGCAATCGCAGACAA TTTAGTCTCTAGAACTGCTGATAATAAATTTGAAGAAGAA
SP123 amino acid (SEQ ID NO:218)
WEVETPQSITNQEQARTENQWETEEAPKEEAPKTEESPKEEPKSEVKPTDDTLPKVEEGKEDSAEPA PVEEVGGEVESKPEEKVAVKPESQPSDKPAEESKVEQAGEPVAPREDEKAPVEPEKQPEAPEEEKAVEE TPKQEESTPDTKAEETVEPKEETVNQSIEQPKVETPAVEKQTEPTEEPKVEQAGEPVAPREDEQAPTAP VEPEKQPEVPEEEKAVEETPKPEDKIKGIGTKEPVDKΞELNNQIDKASSVSPTDYSTASYNALGPVLET AKGVYASEPVKQPEVNSETNKLKTAIDALNVDKTELNNTIADAKTKVKEHYSDRSWQNLQTEVTKAEKV AANTDAKQΞEVNEAVEK TATIEKLVELSEKPILT TSTDKKILEREAVAKYT ENQNKTKIKΞITAEL KKGEEVINTW TDDKVTTETISAAFKNLEYYKEYTLSTTMIYDRGNGEETET ENQNIQ DLKKVE K NIKRTD IKYENGKETNESLITTIPDDKSNYYLKITΞ NQKTTLLAVKNIEETTVNGTPVYKVTAIADN LVSRTADNKFEEE
SP124 amino acid (SEQ ID NO:219)
AACACCTGTATATAAAGTTACAGCAATCGCAGACAATTTAGTCTCTAGAACTGCTGATAATAAATTTGA AGAAGAATACGTTCACTATATTGAAAAACCTAAAGTCCACGAAGATAATGTATATTATAATTTCAAAGA ATTAGTGGAAGCTATTCAAAACGATCCTTCAAAAGAATATCGTCTGGGACAATCAATGAGCGCTAGAAA TGTTGTTCCTAATGGAAAATCATATATCACTAAAGAATTCACAGGAAAACTTTTAAGTTCTGAAGGAAA ACAATTTGCTATTACTGAATTGGAACATCCATTATTTAATGTGATAACAAACGCAACGATAAATAATGT Table 1 94
GAATTTTGAAAATGTAGAGATAGAACGTTCTGGTCAAGATAATATTGCATCATTAGCCAATACTATGAA AGGTTCTTCAGTTATTACAAATGTCAAAATTACAGGCACACTTTCAGGTCGTAATAATGTTGCTGGATT TGTAAATAATATGAATGATGGAACTCGTATTGAAAATGTTGCTTTCTTTGGCAAACTACACTCTACAAG TGGAAATGGCTCTCATACAGGGGGAATTGCAGGTACAAACTATAGAGGAATTGTTAGAAAAGCATATGT TGATGCTACTATTACAGGAAACAAAACACGCGCCAGCTTGTTAGTTCCTAAAGTAGATTATGGATTAAC TCTAGACCATCTTATTGGTACAAAAGCTCTCCTAACTGAGTCGGTTGTAAAAGGTAAAATAGATGTTTC AAATCCAGTAGAAGTTGGAGCAATAGCAAGTAAGACTTGGCCTGTAGGTACGGTAAGTAATTCTGTCAG CTATGCTAAGATTATCCGTGGAGAGGAGTTATTCGGCTCTAACGACGTTGATGATTCTGATTATGCTAG TGCTCATATAAAAGATTTATATGCGGTAGAGGGATATTCGTCAGGTAATAGATCATTTAGGAAATCTAA AACATTTACTAAATTAACTAAAGAACAAGCTGATGCTAAAGTTACTACTTTCAATATTACTGCTGATAA ATTAGAAAGTGATCTATCTCCTCTTGCAAAACTTAATGAAGAAAAAGCCTATTCTAGTATTCAAGATTA TAACGCTGAATATAACCAAGCCTATAAAAATCTTGAAAAATTAATACCATTCTACAATAAAGATTATAT TGTATATCAAGGTAATAAATTAAATAAAGAACACCATCTAAATACTAAAGAAGTTCTTTCTGTTACCGC GATGAACAACAATGAGTTTATCACAAACCTAGATGAAGCTAATAAAATTATTGTTCACTATGCGGACGG TACAAAAGATTACTTTAACTTGTCTTCTAGCAGTGAAGGTTTAAGTAATGTAAAAGAATATACTATAAC TGACTTAGGAATTAAATATACACCTAATATCGTTCAAAAAGATAACACTACTCTTGTTAATGATATAAA ATCTATTTTAGAATCAGTAGAGCTTCAGTCTCAAACGATGTATCAGCATCTAAATCGATTAGGTGACTA TAGAGTTAATGCAATCAAAGATTTATATTTAGAAGAAAGCTTCACAGATGTTAAAGAAAACTTAACAAA CCTAATCACAAAATTAGTTCAAAACGAAGAACATCAACTAAATGATTCTCCAGCTGCTCGTCAAATGAT TCGTGATAAAGTCGAGAAAAACAAAGCAGCTTTATTACTAGGTTTAACTTACCTAAATCGTTACTATGG AGTTAAATTTGGTGATGTTAATATTAAAGAATTAATGCTATTCAAACCAGATTTCTATGGTGAAAAAGT TAGCGTATTAGACAGATTAATTGAAATCGGTTCTAAAGAGAACAACATTAAAGGTTCACGTACATTCGA CGCATTCGGTCAAGTA
SP124 amino acid (SEQ ID NO:220)
TPVYKVTAIADNLVSRTADNKFEEEYVHYIEKPKVHEDNVYYNFKELVEAIQNDPSKEYRLGQSMSARN WPNGKΞYITKEFTGKLLΞSEGKQFAITELEHPLFNVITNATINNVNFE VEIERSGQDNIASLANTMK GSSVITNVKITGTLSGRNNVAGFVNNMNDGTRIENVAFFGKLHSTSGNGSHTGGIAGTNYRGIVRKAYV DATITGNKTRASLLVPKVDYG TLDH IGTKAL TEΞWKGKIDVSNPVEVGAIASKTWPVGTVSNSVS YAKIIRGEELFGSNDVDDSDYAΞAHIKDLYAVEGYSSGNRΞFRKSKTFTKLTKEQADAKVTTFNITADK LESDLSPLAKLNEEKAYSSIQDYNAEYNQAYKNLEKLIPFYNKDYIVYQGNKLNKEHH NTKEVLSVTA MNNNEFITNLDEANKIIVHYADGTKDYFNLSSSSEGLSNVKEYTITDLGIKYTPNIVQKDNTTLVNDIK SILESVELQSQTMYQHLNRLGDYRVNAIKDLYLEΞSFTDVKENLTNLITKLVQNEEHQLNDSPAARQMI RDKVΞKNKAALLLGLTYLNRYYGVKFGDVNIKE MLFKPDFYGEKVSVLDRLIEIGSKEISΓNIKGSRTFD
AFGQV
SP125 nucleotide (SEQ ID NO:221)
ATTAGACAGATTAATTGAAATCGGTTCTAAAGAGAACAACATTAAAGGTTCACGTACATTCGACGCATT CGGTCAAGTATTGGCTAAATATACTAAATCAGGTAATTTAGATGCATTTTTAAATTATAATAGACAATT GTTCACAAATATAGACAATATGAACGATTGGTTTATTGATGCTACAGAAGACCATGTCTACATCGCAGA ACGCGCTTCTGAGGTCGAAGAAATTAAAAATTCTAAACATCGTGCATTCGATAATTTAAAACGAAGTCA CCTTAGAAATACTATACTCCCACTACTGAATATTGATAAAGCACATCTTTATTTAATTTCAAATTATAA TGCAATTGCCTTTGGTAGTGCAGAGCGATTAGGTAAAAAATCATTAGAAGATATTAAAGATATCGTTAA CAAAGCTGCAGATGGTTATAGAAACTATTATGATTTCTGGTATCGTCTAGCGTCTGATAACGTTAAACA ACGACTACTAAGAGATGCTGTTATTCCTATTTGGGAAGGTTATAACGCTCCTGGTGGATGGGTTGAAAA ATATGGCCGCTATAATACCGACAAAGTATATACTCCTCTTAGAGAATTCTTTGGTCCTATGGATAAGTA TTATAATTATAATGGAACAGGAGCTTATGCTGCTATATATCCTAACTCTGATGATATTAGAACTGATGT AAAATATGTTCATTTAGAAATGGTTGGTGAATACGGTATTTCAGTTTACACACATGAAACAACACACGT CAACGACCGTGCGATTTACTTAGGTGGCTTTGGACACCGTGAAGGTACTGATGCTGAAGCATATGCTCA GGGTATGCTACAAACTCCTGTTACTGGTAGTGGATTTGATGAGTTTGGTTCTTTAGGTATTAATATGGT ATTTAAACGCAAAAATGATGGGAATCAGTGGTATATTACAGATCCAAAAACTCTAAAAACACGAGAAGA TATTAATAGATATATGAAGGGTTATAATGACACTTTAACTCTTCTTGATGAAATTGAGGCTGAATCTGT GATTTCTCAACAAAATAAAGATTTAAATAGTGCATGGTTCAAAAAAATAGATAGAGAATACCGTGATAA CAATAAATTAAATCAATGGGATAAAATTCGAAATCTAAGTCAAGAAGAGAAAAATGAATTAAATATTCA ATCTGTTAATGATTTAGTTGATCAACAATTAATGACTAATCGCAATCCAGGTAATGGTATCTATAAACC CGAAGCAATTAGCTATAACGATCAATCACCTTATGTAGGTGTTAGAATGATGACCGGTATCTACGGAGG TAATACTAGTAAAGGTGCTCCTGGAGCTGTTTCATTCAAACATAATGCTTTTAGATTATGGGGTTACTA CGGATACGAAAATGGGTTCTTAGGTTATGCTTCAAATAAATATAAACAACAATCTAAAACAGATGGTGA Table 1 95
GTCTGTTCTAAGTGATGAATATATTATCAAGAAAATATCTAACAATACATTTAATACTATTGAAGAATT TAAAAAAGCTTACTTCAAAGAAGTTAAAGATAAAGCAACGAAAGGATTAACAACATTCGAAGTAAATGG TTCTTCCGTTTCATCATACGATGATTTACTGACATTGTTTAAAGAAGCTGTTAAAAAAGATGCCGAAAC TCTTAAACAAGAAGCAAACGGTAATAAAACAGTATCTATGAATAATACAGTTAAATTAAAAGAAGCTGT TTATAAGAAACTTCTTCAACAAACAAATAGCTTTAAAACTTCAATCTTTAAA
SP125 amino acid (SEQ ID NO: 222)
LDRLIEIGSKENNIKGSRTFDAFGQVLAKYTKSGNLDAFLNYNRQLFTNIDNMNDWFIDATΞDHVYIAE RASEVEEIKNSKHRAFDNLKRSHLRNTILPLLNIDKAHLYLISNYNAIAFGSAERLGKKΞLEDIKDIVN KAADGYRNYYDFWYRLASDNVKQRL RDAVIPI EGYNAPGGWVEKYGRYNTDKVYTPLRΞFFGPMDKY YNYNGTGAYAAIYPNSDDIRTDVKYVHLEMVGEYGISVYTHETTHVNDRAIYLGGFGHREGTDAEAYAQ GM QTPVTGSGFDEFGSLGINMVFΪRKNDGNQ YITDPKTLKTREDINRYMKGYNDTLTL DEIEAESV ISQQNKDLNSAWFKKIDREYRDNNKLNQWDKIRNLSQEEKNE NIQSVNDLVDQQLMTNRNPGNGIYKP EAISYNDQSPYVGVRMMTGIYGGNTSKGAPGAVSFKHNAFRLWGYYGYENGFLGYASNKYKQQSKTDGE SVLSDEYIIKKISNNTFNTIEEFKKAYFKEVKDKATKGLTTFEVNGSSVSSYDDLLTLFKΞAVKKDAET LKQEANGNKTVSMNNTVKLKEAVYKKL QQTNSFKTSIFK
SP126 nucleotide (SEQ ID NO:223)
TAAGACAGATGAACGGAGCAAGGTGTTTGACTTTTCCATTCCCTACTATACTGCAAAAAATAAACTCAT TGTCAAAAAATCTGACTTGACTACTTATCAGTCTGTAAACGACTTGGCGCAGAAAAAGGT GGAGCGCA GAAAGGTTCGATTCAAGAGACGATGGCGAAAGATTTGCTACAAAATTCTTCCCTCGTATCTCTGCCTAA AAATGGGAATTTAATCACAGATTTAAAATCAGGACAAGTGGATGCCGTTATCTTTGAAGAACCTGTTTC CAAGGGATTTGTGGAAAATAATCCTGATTTAGCAATCGCAGACCTCAATTTTGAAAAAGAGCAAGATGA TTCCTACGCGGTAGCCATgAAAAAAGATAGCAAGAAATTGAAGAGGCAGTTCGATAAAACCATTCAAAA GTTGAAGGAGTCTGGGGAATTAGACAAACTCATTGAGGAAGCCTTA
SP126 amino acid (SEQ ID NO:224)
KTDERSKVFDFSIPYYTAKNKLIVKKSDLTTYQSVNDLAQKKVGAQKGSIQET AKDLLQNSS VSLPK NGN ITDLKSGQVDAVIFEEPVSKGFVENNPDLAIADLNFEKEQDDSYAVAMKKDSKKLKRQFDKTIQK LKESGELDKLIEEA
SP127 nucleotide (SEQ ID NO:225)
CTGTGAGAATCAAGCTACACCCAAAGAGACTAGCGCTCAAAAGACAATCGTCCTTGCTACAGCTGGCGA CGTGCCACCATTTGACTACGAAGACAAGGGCAATCTGACAGGCTTTGATATCGAAGTTTTAAAGGCAGT AGATGAAAAACTCAGCGACTACGAGATTCAATTCCAAAGAACCGCCTGGGAGAGCATCTTCCCAGGACT TGATTCTGGTCACTATCAGGCTGCGGCCAATAACTTGAGTTACACAAAAGAGCGTGCTGAAAAATACCT TTACTCGCTTCCAATTTCCAACAATCCCCTCGTCCTTGTCAGCAACAAGAAAAATCCTTTGACTTCTCT TGACCAGATCGCTGGTAAAACAACACAAGAGGATACCGGAACTTCTAACGCTCAATTCATCAATAACTG GAATCAGAAACACACTGATAATCCCGCTACAATTAATTTTTCTGGTGAGGATATTGGTAAACGAATCCT AGACCTTGCTAACGGAGAGTTTGATTTCCTAGTTTTTGACAAGGTATCCGTTCAAAAGATTATCAAGGA CCGTGGTTTAGACCTCTCAGTCGTTGATTTACCTTCTGCAGATAGCCCCAGCAATTATATCATTTTCTC AAGCGACCAAAAAGAGTTTAAAGAGCAATTTGATAAAGCGCTCAAAGAACTCTATCAAGACGGAACCCT TGAAAAACTCAGCAATACCTATCTAGGTGGTTCTTACCTCCCAGATCAATCTCAGTTACAA
SP127 amino acid (SEQ ID NO: 226)
CENQATPKETSAQKTIVLATAGDVPPFDYEDKGNLTGFDIEVLKAVDEKLSDYΞIQFQRTAWESIFPGL DSGHYQAAANNLSYTKERAEKYLYSLPIΞNNPLVLVSNKKNPLTSLDQIAGKTTQEDTGTSNAQFI N NQKHTDNPATINFSGEDIGKRILDLA GEFDFLVFDKVSVQKIIKDRGLDLSWD PSADSPS YIIFS SDQKEFKEQFDKALKE YQDGTLEKLSNTYLGGSY PDQSQLQ Table 2 S. pneumoniae Antigenic Epitopes
SP001 ys-1 to Ile-10; eu-13 to Lys-32; Arg-41 to Ile-51; Ser-85 to Glu-97; Ala-159 to His-168; Val-309 to Thr-318; Val-341 to Asn-352; Asn-415 to et-430; Phe-454 to Asn-464; Ser-573 to Gly-591; Asn-597 to Thr-641; and Asn-644 to Ala-664.
SP004
Thr-9 to Thr-24; Ile-29 to Ala-48; Thr-49 to Val-56; Val-286 to Val-
312;
Pro-316 to Glu-344; Val-345 to Ile-367; Gln-368 to Val-399; Ser-400 to
Glu-431; Asn-436 to Ala-457; Ile-467 to Ala-498; and Thr-499 to Glu-
540.
SP006
Glu-1 to Lys-13; Pro-24 to Gly-36; Val-104 to Thr-112; Ala-118 to Asn- 130; Trp-137 to Ala-146; Ser-151 to Ile-159; Ile-181 to Leu-188; and Pro-194 to Tyr-202.
SP007
Gly-1 to Asn-7; Tyr-24 to Gln-34; His-47 to Phe-55; Ser-60 to Ala-67; Ala-122 to Leu-129; Leu-221 to Lys-230; Val-236 to Phe-256; and Asp-271 to Gly-283; and Leu-291 to Asp-297.
/ SP008
Leu-4 to Lys-17; Gln-24 to Leu-32; Asp-60 to Ser-66; Ser-70 to Asp-76; Ala-276 to Lys-283; Asn-304 to Lys-311; and Thr-429 to Pro-437.
SP009
Thr-4 to Glu-11; Leu-50 to Asp-60; Ile-102 to Trp-123; and Ser-138 to Ile-157.
SP010
Phe-34 to Gly-41; Asp-44 to Lys-50; Leu-172 to Val-186; Leu-191 to Val- 198; Ser-202 to Ile-209; and Val-213 to Leu-221.
SP011
Asn-2 to Thr-10; Asp-87 to Ala-102; Tyr-125 to Glu-132; Thr-181 to Tyr-
189; Arg-217 to Thr-232; Asn-257 to Lys-264; Pro-271 to Ser-278; Tyr-
317 to Ala-325; Glu-327 to Pro-337; and Thr-374 to Val-381.
SP012
Gly-1 to Lys-19; Phe-34 to Tyr- 41; Leu-109 to Lys-126; and Leu-231 to Glu-247.
SP013
Ala-1 to Lys-12; Ile-42 to Pro-53; Leu-138 to Lys-146; Ile-205 to Lys- 217; Ser-235 to Ile-251; and Ser-261 to Tyr-272.
SP014
Gly-1 to Val-16; Leu-35 to Leu-44; Asp-73 to Asp-81; Ile-83 to Asp-92; Glu-145 to Ile-153; Phe-188 to Asn-196; Ser-208 to Phe-215; Ile-224 to Leu-231; and Asn-235 to Ala-243.
SP015
Ser-1 to Pro-16; Asn-78 to Glu-88; Ala-100 to Val-108; Ala-122 to Thr- 129; Thr-131 to Ser-137; Leu-201 to Ser-220; and Gly-242 to Val-251. Table 2 S. pneumoniae Antigenic Epitopes
SP016
Gly-1 to Glu-20; Thr-30 to Val-38; Gln-94 to Asn-105; Lys-173 to Pro- 182; Gly-189 to Arg-197; Ser-207 to Val-224; Pro-288 to Leu-298; Ala- 327 to Ala-342; and Ser-391 to Ala-402.
SP017
Ser-1 to Thr-12; Ala-36 to Tyr-45; Gln-48 to Ile-54; Lys-59 to Lys-76; Tyr-113 to Leu-138; and Phe-212 to Asp-219.
SP019
Val-97 to Glu-117; Asp-163 to Leu-169; Thr-182 to Thr-191; and Lys-241 to Ser-250.
SP020
Asn-18 to Lys-25; Thr-47 to Glu-60; Trp-75 to Val-84; Gly-102 to Val- 110; Pro-122 to Ala-131; and Glu-250 to Pro-258.
SP021
Serl to Asp-8; Val-44 to Asp-54; Ala-117 to Val-125; Thr-165 to Thr- 173; and Glu-180 to Pro-189.
SP022 /
Phe-5 to Lys-13; Thr-20 to Ser-36; Glu-59 to Lys-81; Tyr-85 to Gly-93; Trp-94 to Trp-101; and Thr-195 to Trp-208.
SP023
Gln-45 to Glu-59; Asp-69 to Pro-85; Lys-111 to Asn-121; Pro-218 to Ala- 228; and Glu-250 to Asn-281.
SP025
Gln-14 to Thr-20; Gly-27 to Phe-33; Gly-63 to Glu-71; and Ile-93 to Phe-102.
SP028
Asp-171 to Pro-179; Tyr-340 to Glu-350; Pro-455 to Tyr-463; and Asp-474 to Pro-480.
SP030
Leu-22 to Leu-37; Trp-81 to Ala-90; Phe-101 to Ala-106; Thr-124 to Tyr- 130; and Asn-138 to Glu-144.
SP031
Asp-8 to Val-16; Gly-27 to Thr-35; Gly-178 to Asp-195; Thr-200 to Asp209; Trp-218 to Leu-224; and Lys-226 to Asp-241.
SP032
Ser-9 to Asp-28; Phe-31 to Val-40; Gly-42 to Arg-50; Ile-52 to Leu-60; Asp-174 to Phe-186; Leu-324 to Met-333; and Thr-340 to Asn-347.
SP033
Gln-2 to Ile-13; Phe-46 to Ile-53; and Asp-104 to Thr-121.
SP034
Glu-36 to Gly-43; Ala-188 to Asp-196; Trp-313 to Gly-320; and Leu-323 to Leu-329. Table 2 S. pneumoniae Antigenic Epitopes
SP035
Arg-19 to Asp-36, Asp-47 to Val-57, Asn-134 to Thr-143, Asp-187 to Arg- 196, and Glu-222 to Ser-230
SP036
Arg-10 to Arg-17, Lys-29 to Ser-39, Ser-140 to Ala-153, Arg-158 to Tyr- 169, Asp-175 to Ala-183, Gly-216 to Asn-236, Ala-261 to Leu-270, Arg- 282 to Phe-291, and Thr-297 to Ala-305, Pro-342 to Gln-362, Phe-455 to Asp-463, Hιs-497 to Thr-511, Ala-521 to Gly-529, Ile-537 to Val-546, Ile-556 to Ala-568, Pro-581 to Ser-595, Glu-670 to Ala-685, Ser-696 to Ala-705 and Leu-782 to Ser-791
SP038
Glu-61 to Pro-69, Phe-107 to Ala-115 Leu-130 to Tyr-141, Ala-229 to G u-237, Ser-282 to Asn-287 Ma-330 to Glu-338, and Tyr-387 to Glu- 393
SP039
Ser-28 to Asp-35 Pro-88 to Pro-96, Leu-125 to Arg-135, Phe-149 to Leu- 157, Gln-246 to Val-254 ^la-357 to Thr-362 Gly-402 to Lys-411, and Leu-440 to Pro-448
SP040
Thr-21 to Ile-30 Hιs-54 to Gln-68 Arg-103 to Leu-117, and Thr-127 to Leu- 136
SP041
Gly-36 to Asp-49, Leu-121 to Val-128 and Ala-186 to Ile-196
SP042
Gly-11 to Arg-19, Ile-23 to Lys-31 His-145 to Asn-151, Gln-159 to Asp-
166, Ile-175 to Asp-181 Gly-213 to Tyr-225, Ile-283 to Val-291, Pro-
329 to Glu-364 Arg-372 to Ser-386 Thr-421 to Phe-430 Leu-445 to Val-
453, Ile-486 to Ala-497 Asp-524 to Ala-535, Hιs-662 to Gly-674 and Hιs-679 to Gln-702
SP043
Lys-2 to Asp-12, Val-58 to Asn-68 Ser-87 to Asp-95 and Asp-102 to Lys -117
SP044
Gln-3 to Lys-11, Asp-37 to Tyr-52, Glu-171 to Leu-191, Hιs-234 to Asn- 247, and Asn-283 to Ala-291
SP045
Tyr-52 to Ile-63, Asp-212 to Gln-227, Ser-315 to Thr-332, Leu-345 to Phe-354, Asp-362 to Val-370, Thr-518 to Asn-539, Ala-545 to Lys-559, and Val-601 to Pro-610
SP046
Gln-9 to Ala-18, Glu-179 to Lys-186, Lys-264 to Glu-271, Gly-304 to Glu-17, Ser-503 to Asn-511, Asn-546 to Thr-553, and Asn-584 to Asp-591
SP048 Table 2 S. pneumoniae Antigenic Epitopes
Tyr-4 to Asp-25; Lys-33 to Val-70; Asp-151 to Thr-170; Asp-222 to Val-257; Thr-290 to Phe-301; and Gly-357 to Val-367.
SP049
Ala-23 to Arg-37; Tyr-85 to Gln-95; Glu-106 to Ile-118; Arg-131 to ILE-144; Gly-150 to Ser-162; and Ala-209 to Asp-218.
SP050
Asp-95 to Glu-113; Gly-220 to Gly-228; Asn-284 to Glu-295; Thr-298 to Val-315.
SP051
Lys-16 to Glu-50; Lys-57 to Asn-104; Ser-158 to Trp-173; Asp-265 to Pro-279; Val-368 to Tyr-386; Glu-420 to Ile-454; Pro-476 to Ile-516; Phe-561 to Gly-581; Thr-606 to Gly-664; and Glu-676 to Val-696.
SP052
Asn-41 to Tyr-60; Phe-80 to Glu-103; Ala-117 to Val-139; Ile-142 to Leu-155; Val-190 to Lys-212; Glu-276 to Phe-283; Arg-290 to Ser-299; Leu-328 to Val-351; Gly-358 to Thr-388; Glu-472 to Ala-483; Val-533 to Asn-561; Asp-595 to Val-606; Glu-609 to Val-620; Glu-672 to Ser- 691. ,
SP053
Ala-62 to Val-101; Thr-147 to Leu-174; Lys-204 to Val-216; Gln-228 to Val-262; Ser-277 to Gly-297; Thr-341 to Glyn-368; Thr-385 to Ala- 409; Thr-414 to Ser-453; Asn-461 to Leu-490; Glu-576 to Thr-625; Gly-630 to Arg-639; and Asp-720 to Leu-740.
SP054
Glu-7 to Val-28; and Tyr-33 to Glu-44.
SP055
Pro-3 to Val-18; Thr-21 to Lys-53; Val-84 to Lys-99; Ile-162 to Val- 172; and Val-204 to Ser-241.
SP056
Val-34 to Tyr-41; Leu-47 to Glu-55; and Pro-57 to Gln-66.
SP057
Asp-1 to Val-25; Pro-29 to Ile-80; Asn-96 to Val-145; and Pro-150 to Glu-172.
SP058
Ala-64 to Thr-70; Leu-82 to His-138; and Val-228 to Asn-236.
SP059
Val-10 to Thr-24; Ser-76 to Pro-102; Ser-109 to Ile-119; Ser-124 to Val-130; Thr-186 to Ile-194; and Asn-234 to Ser-243.
SP060
Leu-70 to Arg-76; and Val-79 to Ile-88.
SP062
Glu-14 to Lys-28; Ser-32 to Lys-46; and Glu-66 to Thr-74. Table 2 S. pneumoniae Antigenic Epitopes
SP063
Ile-10 to Val-25; Val-30 to Thr-40; Asp-44 to Pro-54; Asn-57 to Val- 63; Pro-71 to Val-100; and Thr-105 to Thr-116.
SP064
Pro-12 to Leu-32; Val-40 to Leu-68; Asp-95 to Ala-125; Ser-164 to Glu-184; Ser-314 to Glu-346; Asn-382 to Val-393; Leu-463 to Gln-498; Asn-534 to Lys-548; and Lys-557 to Gly-605.
SP065
Asn-2 to Ile-12; Ala-39 to Thr-61; and His-135 to Ala-155.
SP067
Gly-1 to Thr-13; Asp-203 to Asn-218; and Gly-240 to Asp-253.
SPO68
Ser-2 to Ser-12; Val-17 to Gln-26; and Lys-54 to Cys-67.
SP069
Ser-32 to Thr-41; Pro-66 to Glu-80; Thr-110 to Val-122; and Val-147 to Thr-180.
SP070
Lys-6 to Tyr-16; Gln-19 to Ile-27; Arg-50 to Ala-58; Leu-112 to Val- 128; Ile-151 to Asn-167; Leu-305 to Phe-321.
SP071
Gln-92 to Asn-158; Gln-171 to Gln-188; Val-204 to Val-240; Thr-247 to Ala-273; Glu-279 to Thr-338; Pro-345 to Glu-368; Asn-483 to Lys-539; Val-552 to Aia-568; Glu-575 to Ser-591; Ser-621 to Gly-640; Gln-742 to Gly-758.
SP072
Val-68 to Tyr-81; Tyr-86 to Val-121; Leu-127 to Gly-140; Gly-144 to Ala-155; Gln-168 to Val-185; Asp-210 to Try-241; Glu-246 to Thr-269; Lys-275 to Tyr-295; Gly-303 to Pro-320; Arg-327 to Ile-335; Thr-338 to Thr-364; Tyr-478 to Phe-495; and Tyr-499 to Arg-521.
SP073
Glu-37 to Val-45; Glu-55 to Val-68; Thr-104 to Thr-119; Ile-127 to Tyr-135; Asn-220 to Ile-232; Thr-237 to Ala-250; Ser-253 to Ala-263; Glu-284 to Ile-297; and Met-438 to Asn-455.
SP074
Gly- 2 to Ala-12; Gly-96 to Ile-110; and Thr-220 to Phe-239.
SP075
?he-33 to Tyr-42; Gln-93 to Gly-102; and Val-196 to Asp-211.
SP076
3er-64 to Leu-76; and Phe-81 to Ala-101.
SP077
Asp-1 to Glu-12; Tyr-26 to Val-36; and Val-51 to Try-62. Table 2 S. pneumoniae Antigenic Epitopes
SP078
Ala-193 to Ile-208; Tyr-266 to Asn-275; Glu-356 to Leu-369; Ala-411 to Gly-422; Ser-437 to Pro-464; Thr-492 to Glu-534; and Glu-571 to Gln-508.
SP079
Gly-11 to Leu-20; Lys-39 to Leu-48; Leu-72 to Val-85; Asn-147 to Ser- 158; Ile-178 to Asp-187; Tyr-189 to Gln-201; and Leu-203 to Ala-216
SPO80
Ser-2 to Glu-12; Gln-42 to Ala-51; Ala-116 to Ser-127; Phe-131 to Asp-143; and Ile-159 to Ile-171.
SP081
Gln-2 to Leu-9; Gln-49 to Cys-57; Ile-108 to Val-131; Gly-134 to Leu- 145; and Trp-154 to Cys-162.
SP082
Ile-101 to Ser-187; Gly-191 to Asn-221; Arg-225 to Arg-236; Tyr-239 to Leu-255; and Gly-259 to Arg-268.
SP083
Ser-28 to Asp-7,0.
SP084
Leu-42 to GIn-66; Thr-69 to Lys-81; Glu-83 to Arg-92; and Gly-98 to Asn-110.
SP085
Gln-2 to Val-22; and Ser-45 to Glu-51.
SP086
Leu-18 to Gln-65; and Lys-72 to Val-83.
SP087
Ser-45 to Leu-53; and Thr-55 to Gln-63
SP088
Pro-8 to Ile-16; Leu-25 to Trp-33; Tyr-35 to Gln-43; Leu-51 to Val-59;
Val-59 to Arg-67; Thr-55 to Tyr-63; Asn-85 to Gly-93; Thr-107 to
Leu-115;
Leu-115 to Trp-123; Ala-121 to Thr-129; Tyr-153 to Ala-161; His-176 to
Gly-184; Tyr-194 to Ala-202; Ala-217 to Gly-225; and Asn-85 to Gly-93.
SP089
Trp-43 to Ala-51; Gln-68 to Phe-76; Val-93 to Gin-101; Phe-106 to Phe-114; Lys-117 to Lys-125; Trp-148 to Phe-156; Glu-168 to Gln-176; Ile-193 to Tyr-201; Lys-203 to Lys-211; Glu-212 to Gln-220; Ile-237 too Tyr-245; Lys-247 to Lys-255; Glu-256 to Gln-264; Met-275 to Gly-283; Lys-286 to Gly-294; Trp-292 to Glu-300; Asp-289 to Thr-297; Tyr-315 to Ser-323; Asp-334 to Lys-342; Pro-371 to Arg-379; Arg-485 to Asn-493; Lys-527 to Arg-535; Phe-537 to Met-545; and Tyr-549 to Glu-557.
SP090 Table 2 S. pneumoniae Antigenic Epitopes
Phe-2 to Gin- 10; Gln-13 to Lys-21; Tyr-19 to Glu-27; Tyr-39 to Met-47; Pro-65 to Leu-73; Tyr-121 to His-129; Lys-147 to Ile-155; Gly-161 to Lys-169; Gly-218 to Trp-226; Asp-230 to Thr-238; Tyr-249 to Ala-257; and Ala-272 to Gly-280.
SP091
Ser-19 to Ser-27; Asn-25 to Thr-33; Val-51 to Gln-59; Asn-75 to Asn-83; Ile-103 to Trp-111; Tyr-113 to Ala-121; Leu-175 to Asn-183; Glu-185 to Trp-193; Ala-203 to Tyr-211; Val-250 to Phe-258; Asn-260 to Thr-268; Ser-278 to Asp-286; Tyr-305 to Leu-313; Asn-316 to Gly-324; Asn-374 to Asp-382; Asn-441 to Gly-449; and Ser-454 to Gln-462.
SP092
Arg-95 to Glu-103; Ala-216 to Val-224; Leu-338 to Glu-346; Pro-350 to
Ala-358; Pro-359 to Ala-367; ?ro-368 to Ala-376; Pro-377 to Ala-385;
Pro-386 to Ala-394; Pro-395 to Ala-403; Pro-350 to Ala-358; Gln-414 to
Lys-422; Pro-421 to Asn-429; Trp-465 to Tyr-473; Phe-487 to Tyr-495;
Asn-517 to Gly-525; Trp-586 to Tyr-594; Phe-608 to Tyr-616; and Asp-630 to Gly-638.
SP093
Gln-30 to Ile-38; Gln-52 to Val-60; Ala-108 to Hxs-116; Tyr-133 to Glu-141; Tyr-19,2 to Ala-200; and ?he-207 to Ser-215.
SP094
Ala-87 to Val-95; Leu-110 to Cys-118; Gln-133 to Leu-141; Ser-185 to Leu-193; Ile-195 to Gly-203; Asp-206 to Gln-214; Ser-211 to Gly- 219; Ile-241 to Thr-249.
SP095
Arg-1 to Gln-9; Phe-7 to Asn-15; Thr-21 to Asn-30; Leu-46 to Phe-54; and Ξer-72 to Met-80.
SP096
Gly-29 to Ile-37; Glu-52 to Ser-60; and Leu-64 to Gly-72.
SP097
Ala-11 to Thr-19; Glu-53 to Glu-61; Ser-91 to Lys-99; Thr-123 to Gln-131; and Gly-209 to Lys-217.
SP098
Thr-3 to Ser-11; Gly-38 to ?he-46; Tyr-175 to Asn-183; Met-187 to Cys-195; Gln-197 to Leu-205; Tyr-307 to Gln-315; Gly-318 to Tyr-326; Asn-348 to Val-356; Lys-377 to Pro-385; and Leu-415 to Val-423.
SP099
Arg-19 to Gly-27; Asp-76 to Ser-84; Val-90 to Lys-98; Phe-165 to Val-173; Leu-237 to Pro-245.
SP100
His-111 to Gln-119; Ser-141 to His-149; Asp-154 to Ser-162; Gln-158 to Gln-166; Asp-154 to Gln-166; Lys-180 to Gln-188; and Ser-206 to Gln-214.
SP101 Table 2 S. pneumoniae Antigenic Epitopes
Glu-23 to Glu-31; Glu-40 to Val-48; Gln-50 to Ser-53; Thr-61 to Ile-69; Leu-82 to Ile-90; Ala-108 to Leu-116; Gln-121 to Pro-129; and Leu-130 to Thr-138.
SP102
Asp-32 to His-40; Arg-48 to Lys-56; and Asp-102 to Thr-110.
SP103
Arg-5 to Gln-13; Gln-22 to Leu-30; Arg-151 to Gln-159; Arg-167 to
Gln-175; Pro-189 to Glu-197; Gly-207 to Leu-215; Ser-219 to Gln-227;
Ser-233 to Ser-241; Pro-255 to Asp-264; Lys-272 to Gly-280; Ser-318 to Val-326; Thr-341 to Asp-351; Asn-356 to Thr-364; Val-370 to
Tyr-378;
Ile-379 to Gln-387; and et-435 to Tvr-443.
SP105
Asn-28 to Pro-36; Thr-77 to ?he-85; Arg-88 to Val-96; Gly-107 to Phe-115; Asp-169 to Asp-177; His-248 to Ser-256; and Ser-274 to Ala-282.
SP106
Val-10 to Thr-/18; Ile-62 to Tyr-70; Ile-71 to Pro-79; Lys-86 to Gln-94; Lys-100 to Thr-108; Phe-132 to Leu-140; and Asp-145 to Arg-153.
SP107
Asp-33 to Val-41; and Arg-63 to Gln-71.
SP108
Lys-9 to Gln-17; Leu-44 to Ser-52; Ser-63 to Phe-71; Tyr-109 to Ser-117; Ile-183 to Ile-191; Pro-194 to Leu-202; Gly-257 to Gln-265; Ala-323 to Thr-331; and Leu-381 to Tyr-389.
SP109
Asn-2 to Gln-10; Ala-65 to Lys-73; Leu-76 to Glu-84; Thr-111 to Asp-119; Gln-116 to Tyr-124; Tyr-130 to Val-138; Asp-173 to Gly-181; Asp-196 to Ser-204; Asn-231 to Ser-239; Phe-252 to Ser-260; Phe-270 to Tyr-278; Val-291 to His-299; Asp-306 to Leu-314; and Pro-327 to Gly-335.
SP110
Ser-8 to Glu- 16; Ile-37 to Val-45; Ala-107 to Val-115; and Gly-122 to Thr-130.
SP111
Asp-19 to Glu-28; Leu-43 to Ala-51; Asn-102 to Phe-110; Gln-133 to Ξer-141; Phe-162 to Asp-170; Tyr-194 to Met-202; and Asp-273 to Ser-281. Table 2 S. pneumoniae Antigenic Epitopes
SP112
Asp-3 to Gln-11, Gly-21 to Ile-29, Ala-46 to Arg-54, Arg-98 to Arg-106, Thr-114 to Val-122, Gln-133 to Asn-141, and Leu-223 to Thr-231
SP113
Asn-19 to Gly-27, Arg-54 to Ser-62, Val-69 to Gln-77, Ser-117 to
Asn-125, Gly-164 to Leu-172, Tyr-193 to Ser-201, Cys-303 to Phe-311,
His-315 to Ile-323, Arg-341 to Cys-349, Ile-347 to Ser-355, Arg-403 to Phe-411, Gln-484 to Pro-492, Ser-499 to Leu-507, Ile-541 to
Thr-549
Asn-622 to Ile-630, and Glu-645 to Gly-653
SP114
Gly-17 to Leu-25, Hιs-40 to Gln-48, Arg-49 to Arg-57, Ile-65 to Pro-73 ,
Asn-101 to Asp-Ill, Gly-128 to Cys-136 Phe-183 to Thr-191 and Pro-268 to le-276
SP115
Met-8 to Ser-16, Tyr-24 to Leu-32, Cys-68 to Leu-76 Ser-100 to
Pro-108, Thr-193 to Thr-201 Gly-238 to Pro-250, Thr-280 to Phe-288,
Pro-303 to Asrv312, Trp-319 to Leu-328, Leu-335 to Leu-344, Lys-395 to Ala-403, Asn-416 to Gln-424, Tyr-430 to Ser-438, Val-448 to
Leu-456, Leu-460 to Thr-468, Pro-502 to Thr-510, Lys-515 to
Ile-524, Gln-523 to Hιs-532, Tyr-535 to Thr-543, Ser-559 to
Pro-567, Thr-572 to Asn-580,
Val-594 to Arg-602, Arg-603 to Asn-611, Thr-620 to Trp-628, and Tyr-644 to Arg-653
SP117
Ala-6 to Gly-14, Ile-19 to Thr-27, Thr-99 to Leu-107, Ser-117 to Asp-125 Hιs-131 to Val-139 Ile-193 to Gly-201 and Val-241 to Gln-249
SP118
Ser-8 to Trp-23, Hιs-46 to Ala-54, Asn-93 to Gly-101, Val-100 to Ξer-108, Arg-155 to Asp-163, and Hιs-192 to Leu-200
SP119
Tyr-46 to Lys-54, Ξer-93 to Ser-101, Trp-108 to Asn-116, Val-121 to Glu-129, and Tyr-131 to Gln-139
SP120
Ala-57 to Lys-65, Leu-68 to Glu-76, Thr-103 to Tyr-116, Tyr-122 to Val-130, His-163 to Gly-173, Asp-188 to Ser-196, Ser-222 to Ser-231, Phe-244 to Ser-252, Pro-262 to Tyr-270, Val-283 to Hxs-291, and Asp-298 to Leu-306
SP121
Ser-3 to Ala-11, Asp-13 to Leu-21, Ser-36 to Val-44, and Gln-136 to Met-144
SP122
Asn-28 to Lys-36, Glu-39 to Thr-50, Val-54 to Lys-62, Asn-106 to Leu-114 Phe-159 to Gly-167, Asn-172 to Arg-180, Glu-199 to Asn-207, Table 2 S . pneumoniae Antigenic Epitopes
Lys-230 to His-241; Asn-252 to Gly-263; Met-278 to Ala-287; Thr-346 to Asp-354; Lys-362 to Thr-370; Asp-392 to Asn-405; Asp-411 to Ala-424; Gly-434 to Gly-443; Tyr-484 to Glu-492; Ile-511 to Leu-519; Asn-524 to Asp-538; Glu-552 to Ile-567; Val-605 to Lys-613; Phe-697 to Ala-705; Phe-722 to Leu-730; Leu-753 to Leu-761; Asp-787 to Gln-795; Leu-858 to Asn-866; Ala-892 to Thr-901; Gly-903 to Ile-913; Ile-921 to Asn-931; Asn-938 to Pro-951; Gly-960 to Lys-970; Leu-977 to Asp-985; and Leu-988 to Pro-996.
SP123
Val-4 to Asn-12; Glu-47 to Leu-55; Lys-89 to Glu-100; Ser-165 to Thr-173; Lys-234 to Val-242; Ser-258 to Ser-266; Glu-284 to Asn-292; Tyr-327 to Leu-335; Tyr-457 to Thr-465; Tyr-493 to Glu-501; Thr-506 to Tyr-514; Lys-517 to Thr-525; Asn-532 to Gly-540; and Arg-556 to Glu-564.
SP124 rg-16 to Glu-24; Gln-52 to Arg-60; Asn-69 to Tyr-77; Glu-121 to
Asn-129; Ala-134 to Val-142; Thr-151 to Ala-159; Asn-164 to Glu-172;
His-181 to His-189; Thr-210 to Ala-218; Ser-244 to Val-252; Phe-287 to Tyr-297; Ser-312 to Thr-323; His-433 to Tyr-441; Ser-445 to
Asn-453;
Asn-469 to Thr/-477; Asn-501 to Asn-509; Gln-536 to Ala-547; and
Gln-608 to Asp-621.
SP125
Ser-9 to Asp-21; Ala-28 to Leu-36; Asn-49 to Phe-57; Val-137 to Arg-145; Asn-155 to Leu-163; Glu-183 to Asp-191; Gly-202 to Tyr-210; Pro-221 to Asp-229; Phe-263 to Ala-271; Phe-300 to Gln-308; Asp-313 to Glu-321; Asn-324 to Asp-332; Ile-346 to Asn-354; Asp-362 to Lys-370; Met-402 to Gly-410; Gly-437 to Gly-445; Ser-471 to Glu-483; Gly-529 to Asp-537; Gln-555 to Val-563; and Leu-579 to Lys-587.
SP126
Leu-22 to Thr-30 ; Val- 65 to Leu-73 ; and Thr-75 to Asp- 83 .
S P 127
Glu-2 to Ala-12; Asp-28 to Thr-36; Val-105 to Thr-113; Lys-121 to Thr-129; Trp-138 to Pro-146; Ser-152 to Ile-160; Lys-180 to Asp-188; Leu-194 to Asn-202; and Gly-228 to Thr-236.
Table 3 S . pneumoniae ORF Cloning Primers
Primer Name SEQ ID Sequence RE
SP001A NO: 227 GACTGGATCCTAAAATCTACGACAATAAAAATC Sam HI
SP001B NO: 228 CTGAGTCGACTGGTTGTGCTGGTTGAG Sal I
SP004A NO: 229 GTCAGGATCCAAATTACAATACGGACTATG Sam HI
SP004B NO: 230 CAGTGTCGACTAACTCTAGGTCGGAAAC Sal I
SP006A NO: 231 GACTGGATCCTGAGAATCAAGCTACACCCAAAGAG Bam HI
SP006B NO: 232 AGTCAAGCTTTTGTAACTGAGATTGATCTGG Hind III
SP007A NO: 233 GACTGGATCCTGGTAACCGCTCTTCTCGTAACGCAGC Bam HI
SP007B NO: 234 AGTCAAGCTTTTTCAGGAACTTTTACGCTTCC Hind III
SP008A NO: 235 AGTCAGATCTTGTGGAAATTTGACAGGTAACAGCAAAAAAGCTGC Bgl II
SP008B NO: 236 ACTGAAGCTTTTTTGTTTTTCAAGAATTCATCG Hind III
SP009A NO: 237 GACTGGATCCTGGTCAAGGAACTGCTTCTAAAGAC Bam HI
SP009B NO: 238 AGTCAAGCTTTCACAAATTCGTTGGTGAAGCC Hind III
SP010A NO: 239 GACTGGATCCTAGCTCAGGTGGAAACGCTGGTTCATCC Bam HI
SP010B NO: 240 AGTCAAGCTTATCAACTTTTCCACCTTCAACAACC Hind III
SP011A NO:241 GTCAAGATCTCTCCAACTATGGTAAATCTGCGGATGG Bgl II
SP011B NO: 242 AGTCCTGCAGATCCACATCCGCTTTCATCGGGTTAAAGAAGG Pst I
SP012A NO: 243 GACTGGATCCTGGGAAAAATTCTAGCGAAACTAGTGG Ba HI
SP012B NO: 244 GTCACTGCAGCTGTCCTTCTTTTACTTCTTTGGTTGC Pst I
SP013A NO: 245 GACTGGATCCTGCTAGCGGAAAAAAAGATACAACTTCTGG Bam HI
SP013B NO:246 CTGAAAGCTTTTTTGCCAATCCTTCAGCAATCTTGTC Hind III
SP014A NO: 247 GACTAGATCTTGGCTCAAAAAATACAGCTTCAAGTCC Bgl II
SP014B NO:248/ AGTCCTGCAGGTTTTTGTTTGCTTGGTATTGGTCG Pst I
SP015A NO: 249 GACTGGATCCTAGTACAAACTCAAGCACTAGTCAGACAGAG Bam HI
SP015B NO:250 CAGTCTGCAGTTTCAAAGCTTTTTGTATGTCTTC Pst I
SP016A NO: 251 GACTGGATCCTGGCAATTCTGGCGGAAGTAAAGATGC Bam HI
SP016B NO: 252 AGTCAAGCTTGTTTCATAGCTTTTTTGATTGTTTCG Hind III
SP017A NO: 253 GACTGGATCCTTCACAAGAAAAAACAAAAAATGAAGATGG Bam HI
SP017B NO:254 AGTCAAGCTTATCGACGTAGTCTCCGCCTTC Hind III
SP019A NO: 255 GACTGGATCCGAAAGGTCTGTGGTCAAATAATCTTACC Bam HI
SP019B NO: 256 AGTCAAGCTTAGAGTTAACATGGTGCTTGCCAATAGG Hind III
SP020A NO: 257 GACTGGATCCAAACTCAGAAAAGAAAGCAGACAATGC Bam HI
SP020B NO: 258 AGTCAAGCTTCCAAACTGGTTGATCCAAACCATCTG Hind III
SP021A NO: 259 GACTGGATCCTTCGAAAGGGTCAGAAGGTGCAGACC Bam HI
SP021B NO: 260 AGTCAAGCTTCTGTAGGCTTGGTGTGCCCCAGTTGC Hind III
SP022A NO: 261 CTGAGGATCCGGGGATGGCAGCTTTTAAAAATC Bam HI
SP022B NO:262 CAGTAAGCTTGTTTACCCATTCACCATTACC Hind III
SP023A NO: 263 CAGTGGATCCAGACGAGCAAAAAATTAAG Bam HI
SP023B NO: 264 TCAGAAGCTTGTTTACCCATTCACCATT Hind III
SP025A NO: 265 GACTGGATCCCTGTGGTGAGGAAGAAACTAAAAAG Bam HI
SP025B NO: 266 CTGAGTCGACAATATTCTGTAGGAATGCTTCGAATTTG Sal I
SP028A NO: 267 CTGAGGATCCGACTTTTAACAATAAAACTATTGAAGAG Bam HI
SP028B NO:268 GTCACTGCAGGTTGTCACCTCCAAAAATCACGG Pst I
SP030A NO: 269 GACTGGATCCCTTTACAGGTAAACAACTACAAGTCGG Bam HI
SP030B NO:270 CAGTAAGCTTTTCGAAGTTTGGCTCAGAATTG Hind III
SP031A NO: 271 GACTGGATCCCCAGGCTGATACAAGTATCGCA Bam HI
SP031B NO: 272 CAGTAAGCTTATCTGCAGTATGGCTAGATGG Hind III
SP032A NO: 273 GACTGGATCCGTCTGTATCATTTGAAAACAAAGAAAC Bam HI
SP032B NO: 274 CAGTCTGCAGTTTTACTGTTGCTGTGCTTGTG Pst I
ΞP033A NO:275 ACTGAGATCTTGGTCAAAAGGAAAGTCAGACAGGAAAGG Bgl II
SP033B NO:276 CAGTAAGCTTATTCCTGAGCTTTTTTGATAAAGGTTGCGCA Hind III
SP034A NO: 277 ACTGGGATCCGAAGGATAGATATATTTTAGCATTTGAGAC Bam HI
SP034B NO: 278 AGTCAAGCTTCCATGGTATCAAAGGCAAGACTTGG Hind III
SP035A NO: 279 GTCAGGATCCGGTAGTTAAAGTTGGTATTAACGG Bam HI
SP035B NO:280 AGTCAAGCTTGCAATTTTTGCGAAGTATTCCAAGAG Hind III
SP036A NO: 281 AGTCGGATCCTTCTTACGAGTTGGGACTGTATCAAGC Bam HI Table 3 S. pneumoniae ORF Cloning Primers
Primer Name SEQ ID Sequence RE
SP036B NO:282 AGTCAAGCTTGTTTATTTTTTCCTTACTTACAGATGAAGG Hind III
SP038A NO: 283 AGTCGGATCCTACTGAGATGCATCATAATCTAGGAGC Bam HI
SP038B NO:284 TCAGCTCGAGTTCTTTGACATCTCCATCATAAGTCGC Xho I
SP039A NO: 285 GACTGGATCCGGTTTTGAGAAAGTATTTGCAGGGG Sam HI
SP039B NO: 286 CAGTAAGCTTGGATTTTTTCATGGATGCAATTTTTTTGG Hind III
SP040A NO: 287 GACTGGATCCGACAACATTTACTATCCATACAGTAGAGTCAGC Bam HI
SP040B NO:288 GACTAAGCTTGGCATAAGGTTGCAATTCTGGATTAATTGG Hind III
ΞP041A NO:289 GACTGGATCCGGCTAAGGAAAGAGTGGATG Bam HI
SP041B NO: 290 GACTAAGCTTTTCATTTTTAAATTGACTATGCGCCCG Hind III
SP042A NO: 291 GACTGGATCCTTGTTCCTATGAACTTGGTCGTCACC Bam HI
SP042B NO: 292 CATGAAGCTTATCCTGGATTTTTCCAAGTAAATCT Hind III
SP043A NO: 293 GACTGGATCCTTATAAGGGTGAATTAGAAAAAGG Bam HI
SP043B NO:294 GACTAAGCTTCTTATTAGGATTGTTAGTAGTTG Hind III
SP044A NO: 295 GACTGGATCCGAATGTTCAGGCTCAAGAAAGTTCAGG Bam HI
SP044B NO: 296 GACTAAGCTTTTCCCCTGATGGAGCAAAGTAATACC Hind III
SP045A NO: 297 GACTGGATCCCTTGGGTGTAACCCATATCCAGCTCCTTCC Bam HI
SP045B NO: 298 GACTGTCGACTTCAGCTTGTTTATCTGGGGTTGC Sal I
SP046A NO: 299 GACTGGATCCTAGTGATGGTACTTGGCAAGGAAAACAG 3am HI
SP046B NO: 300 ACTGCTGCAGATCTTTGCCACCTAGCTTCTCATTG Pst I
SP048A NO: 301 GTCAGGATCCTGGGATTCAATATGTCAGAGATGATACTAG 3am HI
SP048B NO: 02 CTAGAAGCTTACGCACCCATTCACCATTATCATTG Hind III
SP049A NO: 303/ GTCAGGATCCGGATAATAGAGAAGCATTAAAAACC Bam HI
SP049B NO:304 AGTCAAGCTTGACAAAATCTTGAAACTCCTCTGGTC Hind III
SP050A NO: 305 GTCAGGATCCAGATTTTGTCGAGGAGTGTCATACC Bam HI
SP050B NO: 306 AGTCAAGCTTTCCCTTTTTACCCTTACGAATCCAGG Hind III
SP051A NO: 307 GACTGGATCCATCTGTAGTTTATGCGGATGAAACACTTATTAC Bam HI
SP051B NO: 308 GACTGTCGACGCTTTGGTAGAGATAGAAGTCATG Sal I
SP052A NO: 309 GACTGGATCCTTACTTTGGTATCGTAGATACAGCCGGC Bam HI
SP052B NO: 310 AGTCAAGCTTTGTTAATTGCGTACCTTCTAAGCGACC Hind III
SP053A NO: 311 GACTGGATCCAGCTAAGGTTGCATGGGATGCGATTCG Sam HI
SP053B NO: 312 GACTGTCGACCTGGGCTTTATTAGTTTGACTAGC Sal I
SP054A NO: 313 CAGTGGATCCCTATCACTATGTAAATAAAGAGA Sam HI
SP054B NO: 314 ACTGAAGCTTTTCTGTCCCTGTTTGAGGCA Hind III
SP055A NO: 315 CAGTGGATCCTGAGACTCCTCAATCAATAACAAA Bam HI
SP055B NO: 316 ACGTAAGCTTATAATCAGTAGGAGAAACTGAACT Hind III
SP056A NO: 317 CAGTGGATCCGGATGCTCAAGAAACTGCGG Bam HI
SP056B NO:318 GACTAAGCTTTTGCCTCTCATTCTTGCTTCC Hind III
SP057A NO: 319 CAGTGGATCCCGACAAAGGTGAGACTGAG Bam HI
SP057B NO: 320 ACGTAAGCTTATTTCTTAATTCAAGTGTTTTCTCTG Hind III
SP058A NO: 321 GACTGGATCCAAATCAATTGGTAGCACAAGATCC Bam HI
SP058B NO: 322 CAGTGTCGACATTAGGAGCCACTGGTCTC Sal I
SP059A NO: 323 CAGTGGATCCCAAACAGTCAGCTTCAGGAAC Bam HI
SP059B NO:324 GACTCTGCAGTTTAATCTTGTCCCAGGTGG Pst I
SP060A NO: 325 GACTGGATCCATTCGATGATGCGGATGAAAAG Bam HI
SP060B NO: 326 GACTAAGCTTCATTTGTCTTTGGGTATTTCGCA Hind III
SP062A NO: 327 CAGTGGATCCGGAGAGTCGATCAAAAGTAG Bam HI
SP062B NO: 328 GTCACTGCAGTTGCTCGTCTCGAGGTTC Pst I
SP063A NO:329 CAGTGGATCCATGGACAACAGGAAACTGGGAC Bam HI
SP063B NO:330 CAGTAAGCTTATTAGCTTCTGTACCTGTGTTTG Hind III
SP064A NO: 331 GACTGGATCCCGATGGGCTCAATCCAACCCCAGGTCAAGTC Bam HI
SP064B NO: 332 GACTCTGCAGCATAGCTTTATCCTCTGACATCATCGTATC Pst I
SP065A NO: 333 GACTGGATCCTTCCAATCAAAAACAGGCAGATGG Bam HI
SP065B NO: 334 GACTAAGCTTGAGTCCCATAGTCCAAGGCA Hind III
SP067A NO: 335 AGTCGGATCCTATCACAGGATCGAACGGTAAGACAACC Bam HI
SP067B NO: 336 ACTGGTCGACTTCTTTTAACTCCGCTACTGTGTC Sal I Table 3 pneumoniae ORF Cloning Primers
Primer Name SEQ ID Sequence RE
SP068A NO: 337 CAGTGGATCCAAGTTCATCGAAGATGGTTGGGAAGTCC Bam HI
SP068B NO: 338 GATCGTCGACCCGCTCCCACATGCTCAACCTT Sal I
SP069A NO: 339 TGACGGATCCATCGCTAGCTAGTGAAATGCAAGAAAG Bam HI
SP069B NO: 340 TGACAAGCTTATTCGTTTTTGAACTAGTTGCTTTCGT Hind III
SP070A NO: 341 GACTGGATCCGCACCAGATGGGGCACAAGGTTCAGGG Bam HI
SP070B NO: 342 TGACAAGCTTAACTTGTAACGAACAGTTCAATCTG Hind III
SP071A NO: 343 GACTAGATCTTTTTAACCCAACTGTTGGTACTTTCC Bgl II
SP071B NO: 344 TGACAAGCTTGTTAGGTGTTACATTTTGACCGTC Hind III
SP072A NO: 345 ACTGAGATCTTTTTAACCCAACTGTTGGTACTTTC Bgl II
SP072B NO: 346 GACTAAGCTTTCTACGATAACGATCATTTTCTTTACC Hind III
SP073A NO: 347 GACTGTCGACTCGTAGATATTTAAGTCTAAGTGAAGCG Sal I
SP073B NO: 348 AGTCAAGCTTGTTAGGTGTTACATTTTGCAAGTC Hind III
SP074A NO: 349 GACTGGATCCCTTTGGTTTTGAAGGAAGTAAG Bam HI
SP074B NO: 350 TGACCTGCAGACGATTTTTGAAAAATGGAGGTGTATC Pst I
SP075A NO: 351 CAGTGGATCCCTACTACCTCTCGAGAGAAAG Bam HI
SP075B NO:352 ACTGAAGCTTTTCGCTTTTTACTCGTTTGACA Hind III
SP076A NO: 353 CAGTGGATCCTAAGGTCAAAAGTCAGACCGCTAAGAAAGTGC Bam HI
SP076B NO:354 CAGTAAGCTTTAGGGTATCCAAATACTGGTTGTTGATG Hind III
SP077A NO: 355 TGACAGATCTTGACGGGTCTCAGGATCAGACTCAGG Bgl II
SP077B NO: 356 TGACAAGCTTCAAAGACATCCACCTCTTGACCTTTG Hind III
SP078A NO: 357 GACTGGATCCTAGAGGCTTTGCCAAATGGTGGGAAGGG Bam HI
SP078B NO: 358/ GTCAGTCGACTTGTTGTAACACTTTTCGAGGTTTGGTACC Sal I
SP079A NO: 359 CAGTGGATCCTCAAAAAGAGAAGGAAAACTTGG 3am HI
ΞP079B NO: 360 CAGTCTGCAGTTTCTTCAACAAACCTTGTTCTTG Pst I
SP080A NO: 361 CAGTGGATCCACGTTCTATTGAGGACCACTT 3am HI
SP080B NO:362 CAGTAAGCTTTTCCTTCTCAGTCAATTCTTTTCC Hind III
SP081A NO: 363 GACTGGATCCCGCTCAAAATACCAGAGGTGTTCAG Bam HI
SP081B NO: 364 GACTAAGCTTAGTACCATGGGTGTGACAGGTTTGAA Hind III
SP082A NO:365 CTGAGGATCCAATTGTACAATTAGAAAAAGATAGC Bam HI
SP082B NO: 366 TGACAAGCTTGCGTTGACTAGGTTCTGCAATGCC Hind III
SP083A NO: 367 GACTGGATCCTCTGACCAAGCAAAAAGAAGCAGTCAATGA Bam HI
SP083B NO: 368 TCAGCAGCTGATCATTGACTTTACGATTTGCTCC Bgl II
SP084A NO: 369 GACTGGATCCGTCCGGCTCTGTCCAGTCCACTTTTTCAGCG Bam HI
ΞP084B NO: 370 TCAGAAGCTTATTTTTTGTTTCCTTAATGCGTT Hind III
SP085A NO: 371 GACTGGATCCGGGACAAATTCAAAAAAATAGGCAAGAGG Bam HI
SP085B NO: 372 GTCAAAGCTTTGGCTCTTTGATTGCCAACAACTG Hind III
SP086A NO: 373 GACTGGATCCTCGCTACCAGCAACAAAGCGAGCAAAAGG 3am HI
ΞP086B NO: 374 GACTAAGCTTACTTTTTTCTTTTTCCACACGA Hind III
SP087A NO: 375 CAGTGGATCCGAACCGACAAGTCGCCCACTATCAAGACT Sam HI
SP087B NO: 376 CTGAAAGCTTTGAATTCTCTTTCTTTTCAGGCT Hind III
SP088A NO: 377 TCGAGGATCCGGTTGTCGGCTGGCAATATATCCCGT 3am HI
SP088B NO: 378 CAGTAAGCTTCCGAACCCATTCGCCATTATAGTTGAC Hind III
SP089A NO: 379 AGTCGGATCCGGCCAAATCAGAATGGGTAGAAGAC Bam HI
SP089B NO:380 TGACCTGCAGCTTCTCATTGATTTTCATCATCAC Pst I
SP090A NO: 381 GACTGGATCCATTTGCAGATGATTCTGAAGGATGG Sam HI
SP090B NO: 382 TCAGCTGCAGCTTAACCCATTCACCATTCTAGTTTAAG Pst I
SP091A NO: 383 GACTGGATCCTGTCGCTGCAAATGAAACTGAAGTAGC Bam HI
SP091B NO: 384 GACTAAGCTTATACCAAACGCTGACATCTACGCG Hind III
SP092A NO: 385 AGTCAGATCTTACGTCTCAGCCTACTTTTGTAAGAGC Bgl II
SP092B NO: 386 GACTAAGCTTAACCCATTCACCATTGGCATTGAC Hind III
SP093A NO: 387 CAGTGGATCCTGGACAGGTGAAAGGTCATGCTACATTTGTG Bam HI
SP093B NO: 388 GACTAAGCTTCAACCATTGAGACCTTGCAACAC Hind III
SP094A NO: 389 GTCAGGATCCGATTGCTCCTTTGAAGGATTTGAGAGAAACC Bam HI
SP094B NO:390 GACTAAGCTTCGATCAAAGATAAGATAAATATATATAAAGT Hind III
SP095A NO:391 GACTGGATCCTAGGTCATATGGGACTTTTTTTCTACAACAAAATAGG Sam HI Table 3 S. pneumoniae ORF Cloning Primers
Primer Name SEQ ID Sequence RE
SP095B NO:392 TGACAAGCTTATCTATCAGCTCATTTAATCGTTTTTG Hind III
SP096A NO: 393 CTGAGGATCCCAACGTTGAGAATTATTTGCGAATG Bam HI
SP096B NO: 394 TGACAAGCTTGAGTCTACAAAAGTAATGTAC Hind III
SP097A NO:395 GTCAGGATCCCTACTATCAATCAAGTTCTTCAGCC Bam HI
SP097B NO: 396 TGACAAGCTTGACTGAGGCTTGGACCAGATTGAAAAG Hind III
SP098A NO: 397 GACTGGATCCGACAAAAACATTAAAACGTCCTGAGG Bam HI
SP098B NO: 398 GACTAAGCTTAGCACGAACTGTGACGCTGGTTCC Hind III
SP099A NO: 399 GACTGGATCCTTCTCAGGAGACCTTTAAAAATATC Sam HI
SP099B NO: 400 GACTAAGCTTGTTGGCCATCTTGTACATACC Hind III
ΞP100A NO: 401 GACTGGATCCAGTAAATGCGCAATCAAATTC Sam HI
SP100B NO:402 AGTCCTGCAGGTATTTAGCCCAATAATCTATAAAGCT Pst I
SP101A NO: 403 CAGTGGATCCTTACCGCGTTCATCAAGATGTC Bam HI
SP101B NO: 404 GACTAAGCTTGCCAGATGTTGAAAAGAGAGTG Hind III
SP102A NO:405 GACTGGATCCGTGGATGGGCTTTAACTATCTTCGTATTCG Bam HI
SP102B NO: 406 AGTCAAGCTTGCTAGTCTTCACTTTCCCTTTCC Hind III
SP103A NO:407 GACTGTCGACACTAAACCAGCATCGTTCGCAGGA Sal I
ΞP103B NO:408 CTGACTGCAGCTTCTTGAAGAAATAATGATTGTGG Pst I
ΞP105A NO: 409 CAGTGGATCCTGACTACCTTGAAATCCCACTT Bam HI
SP105B NO:410 CAGTAAGCTTTTTTTTAAGGTTGTAGAATGATTTCAATC Hind III
SP106A NO: 411 CAGTGTCGACTCGTATCTTTTTTTGGAGCAATGTT Sal I
SP106B NO:412 GACTAAGCTTAAATGTTCCGATACGGGTGATTG Hind III
SP107A NO: 413/ CAGTGGATCCGGACTCTCTCAAAGATGTGAAAG Bam HI
SP107B NO:414 GACTAAGCTTCTTGAGTTTGTCAAGGATTGCTTT Hind III
SP108A NO: 415 CAGTGGATCCCAAGAAATCCTATCATCTCTTCCAGAAG Bam HI
SP108B NO: 416 GACTAAGCTTTTCAGAACTAAAAGCCGCAGCTT Hind III
SP109A NO: 417 GACTGGATCCACGAAATGCAGGGCAGACAG Bam HI
SP109B NO:418 CAGTAAGCTTATCAACATAATCTAGTAAATAAGCGT Hind III
SP110A NO: 419 CAGTGGATCCTGTATAGTTTTTAGCGCTTGTTCTTC Bam HI
ΞP110B NO: 420 GTCAAAGCTTTGATAGAGTGTCATAATCTTCTTTAG Hind III
SP111A NO:421 GACTGGATCCGTGTGTCGAGCATATTCTGAAG Bam HI
SP111B NO: 422 CAGTAAGCTTACTTTTACCATTTCTTTGTTCTGCATC Hind III
SP112A NO: 423 GACTGTCGACGTGTTTGGATAGCATTCAGAATCAGACG Sal I
SP112B NO: 424 CAGTAAGCTTCGGAAGTAAAGACAATTTTTCC Hind III
SP113A NO: 425 CAGTGGATCCGTGCCTAGATAGTATTATTACTCAAAC Bam HI
SP113B NO:426 GACTAAGCTTTTTGCTTATTTCTCTCAATTTTTC Hind III
SP114A NO: 427 CAGTGGATCCCATTCAGAAGCAGACCTATCAAAATC Bam HI
SP114B NO:428 ACTGAAGCTTATGTAATTTTTTAGATTTTTCAATATTTTTCAG Hind III
SP115A NO: 429 AGTCGGATCCTAAGGCTGATAATCGTGTTCAAATG Bam HI
SP115B NO:430 GACTAAGCTTAAAATTAGATAGACGTTGAGT Hind III
SP117A NO: 431 AGTCGGATCCCTGTGGCAATCAGTCAGCTGCTTCC Bam HI
SP117B NO: 432 GACTGTCGACTTTAATCTTGTCCCAGGTGGTTAATTTGCC Sal I
SP118A NO: 433 ACTGGTCGACTTGTCAACAACAACATGCTACTTCTGAG Sal I
SP118B NO: 434 GACTCTGCAGAAGTTTAACCCACTTATCATTATCC Pst I
SP119A NO: 435 ACTGGGATCCTTGTTCAGGCAAGTCCGTGACTAGTGAAC Bam HI
SP119B NO:436 GACTAAGCTTGGCTAATTCCTTCAAAGTTTGCA Hind III
SP120A NO: 437 AGTCGGATCCCTCGCAAATTGAAAAGGCGGCAGTTAGCC Bam HI
SP120B NO: 438 GACTAAGCTTGTAAATAAGCGTACCTTTTTCTTCC Hind III
SP121A NO :439 TCAGGGATCCTTGTCAGTCAGGTTCTAATGGTTCTCAG Bam HI
SP121B NO: 440 AGTCAAGCTTGGCATTGGCGTCGCCGTCCTTC Hind III
SP122A NO: 441 GACTGGATCCGGAAACTTCACAGGATTTTAAAGAGAAG Bam HI
SP122B NO: 442 GACTGTCGACAATCAATCCTTCTTCTGCACTTCT Sal I
SP123A NO: 443 CAGTGGATCCTGTGGTCGAAGTTGAGACTCCTCAATC Sam HI
SP123B NO: 444 GACTAAGCTTTTCTTCAAATTTATTATCAGC Hind III
SP124A NO: 445 AGTCGGATCCAACACCTGTATATAAAGTTACAGCAATCG Bam HI
SP124B NO: 446 GACTGTCGACTACTTGACCGAATGCGTCGAATGTACG Sal I 10
Table 3 S . pneumoniae ORF Cloning Primers
P rime r Name SEQ ID Sequence RE
SP125A NO : 447 CTGAGGATCCATTAGACAGATTAATTGAAATCGG Bam HI SP125B NO:448 GACTGTCGACTTTAAAGATTGAAGTTTTAAAGCT Sal I ΞP126A NO:449 TGACGGATCCTAAGACAGATGAACGGAGCAAGGTG Bam HI SP126B NO:450 CTGAAAGCTTTAAGGCTTCCTCAATGAGTTTGTCT Hind III SP127A NO:451 GACTGGATCCCTGTGAGAATCAAGCTACACCCA Bam HI SP127B NO:452 CTGAAAGCTTTTGTAACTGAGATTGATCTGGGAG Hind III
INDICATIONS RELATING TO A DEPOSITED MICROORGANISM
(PCT Rule \ 3bis)
Figure imgf000113_0001
For International Bureau use onK
D This sneet was received bv the International Bureau on
Autnoπzeα oificer
Figure imgf000113_0002
SINGAPORE
The applicant hereby requests that the furnishing of a sample of a microorganism shall only be made available to an expert. The request to this effect must be filed by the applicant with the International Bureau before the completion of the technical preparations for international publication of the application.
NORWAY
The applicant hereby requests that, until the application has been laid open to public inspection (by the Norwegian Patent Office), or has been finally decided upon by the Norwegian Patent Office without having been laid open to public inspection, the furnishing of a sample shall only be effected to an expert in the art. The request to this effect shall be filed by the applicant with the Norwegian Patent Office not later than at the time when the application is made available to the public under Sections 22 and 33(3) of the Norwegians Patents Act. If such a request has been filed by the applicant, any request made by a third party for the furnishing of a sample shall indicate the expert to be used. That expert may be any person entered on a list of recognized experts drawn up by the Norwegian Patent Office or any person approved by the applicant in the individual case.
AUSTRALIA
The applicant hereby gives notice that the furnishing of a sample of a microorganism shall only be effected prior to the grant of a patent, or prior to the lapsing, refusal or withdrawal of the application, to a person who is a skilled addressee without an interest in the invention (Regulation 3.25(3) of the Australian Patents Regulations).
FINLAND
The applicant hereby requests that, until the application has been laid open to public inspection (by the National Board of Patents and Registration), or has been finally decided upon by the National Board of Patents and Registration without having been laid open to public inspection, the furnishing of a sample shall only be effected to an expert in the an.
ICELAND
The applicant hereby requests that, until the application has been laid open to public inspection (by the Icelandic Patent Office), or has been finally decided upon by the Icelandic Patent Office without having been laid open to public inspection, the furnishing of a sample shall only be effected in the art. Page 2
DENMARK
The applicant hereby requests that, until the application has been laid open to public inspection (by the Danish Patent Office), or has been finally decided upon by the Danish Patent Office without having been laid open to public inspection, the furnishing of a sample shall only be effected to an expert in the art. The request to this effect shall be filed by the applicant with the Danish Patent Office not later than at the time when the application is made available to the public under Sections 22 and 33(3) of the Danish Patents Act. If such a request has been filed by the applicant, any request made by a third party for the furnishing of a sample shall indicate the expert to be used. That expert may be any person entered on a list of recognized experts drawn up by the Danish Patent Office or any person approved by the applicant in the individual case.
SWEDEN
The applicant hereby requests that, until the application has been laid open to public inspection (by the Swedish Patent Office), or has been finally decided upon by the Swedish Patent Office without having been laid open to public inspection, the furnishing of a sample shall only be effected to an expert in the art. The request to this effect shall be filed by the applicant with the International Bureau before the expiration of 16 months from the priority date (preferably on the Form PUT/RO/134 reproduced in annex Z of Volume I of the PCT Applicant's Guide). If such a request has been filed by the applicant, any request has been filed by the applicant, any request made by a third party for the furnishing of a sample shall indicate the expert to be used. That expert may be any person entered on a list of recognized experts drawn up by the Swedish Patent Office or any person approved by the applicant in the individual case.
UNITED KINGDOM
The applicant hereby requests that the furnishing of a sample of a microorganism shall only be made available to an expert. The request to this effect must be filed by the applicant with the International Bureau before the completion of the technical preparations for the Intemationai publication of the application.
NETHERLANDS
The applicant hereby requests that until the date of a grant of a Netherlands patent or until the date on which the application is refused or withdrawn or lapse, the microorganism shall be made available as provided in Rule 31F(1) of the Patent Rules only by the issue of a sample to an expert. The request to this effect must be furnished by the applicant with the Netherlands Industrial Property Office before the date on which the application is made available to the public under Section 22C or Section 25 of the Patents Act of the Kingdom of the Netherlands, whichever two dates occurs earlier.

Claims

What Is Claimed Is:
1 . An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of:
(a) a nucleotide sequence encoding any of the amino acid sequences of the polypeptides shown in Table 1 ; or
(b) a nucleotide sequence complementary to any of the nucleotide sequences in (a).
2. An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a) or (b) of claim 1 wherein said polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues.
3. An isolated nucleic acid molecule comprising a polynucleotide which encodes the amino acid sequence of an epitope-bearing portion of a polypeptide having an amino acid sequence in (a) of claim 1.
4. The isolated nucleic acid molecule of claim 3, wherein said epitope-bearing portion of a polypeptide has an amino acid sequence listed in Table 2.
5. A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of claim 1 into a vector.
6. A recombinant vector produced by the method of claim 5.
7. A method of making a recombinant host cell comprising introducing the recombinant vector of claim 6 into a host cell.
8. A recombinant host cell produced by the method of claim 7.
9. A method of producing a polypeptide encoded by the nucleic acid molecule of claim 1 comprising culturing the host cell of claim 8 under conditions favoring expressing the heterologous polypeptide.
10. A polypeptide produced according to the method of claim 9.
1 1. An isolated polypeptide comprising an amino acid sequence at least 70% identical to a sequence selected from the group consisting of an amino acid sequence of any of the polypeptides described in Table 1.
12. An isolated polypeptide antigen comprising an amino acid sequence of an S. pneumoniae epitope shown in Table 2.
13. An isolated nucleic acid molecule comprising a polynucleotide with a nucleotide sequence encoding a polypeptide of claim 9.
14. An isolated antibody that binds specifically to a polypeptide of claim 1 1.
15. A hybridoma which produces an antibody according to claim 14.
16. A vaccine, comprising: (1) one of more S. pnuemoniae polypeptides selected from the group consisting of a polypeptide comprising an amino acid sequence identified in Table 1 , or a fragment thereof; and
(2) a pharmaceutically acceptable diluent, carrier, or excipient; wherein said polypeptide is present, in an amount effective to elicit protective antibodies in an animal to a member of the Streptococcus genus.
17. A method of preventing or attenuating an infection caused by a member of the Streptococcus genus in an animal, comprising administering to said animal a polypeptide of claim 1 1 , wherein said polypeptide is administered in an amount effective to prevent or attenuate said infection.
18. A method of detecting Streptococcus nucleic acids in a biological sample obtained from an animal involving assaying for one or more nucleic acid sequences encoding Streptococcus polypeptides in a sample comprising: (a) contacting the sample with one or more of the above-described nucleic acid probes, under conditions such that hybridization occurs, and
(b) detecting hybridization of said one or more probes to the one or more Streptococcus nucleic acid sequences present in the biological sample.
19. A method of detecting Streptococcus nucleic acids in a biological sample obtained from an animal, comprising:
(a) amplifying one or more Streptococcus nucleic acid sequences in said sample using polymerase chain reaction, and
(b) detecting said amplified Streptococcus nucleic acid.
20. A kit for detecting Streptococcus antibodies in a biological sample obtained from an animal, comprising (a) a polypeptide of claim 12 attached to a solid support; and
(b) detecting means.
21. A method of detecting Streptococcus antibodies in a biological sample obtained from an animal, comprising (a) contacting the sample with a polypeptide of claim 12; and
(b) detecting antibody-antigen complexes.
PCT/US1997/019422 1996-10-31 1997-10-30 Streptococcus pneumoniae antigens and vaccines WO1998018930A2 (en)

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EP97946850.1A EP0942983B2 (en) 1996-10-31 1997-10-30 Streptococcus pneumoniae antigens and vaccines
JP52066798A JP4469026B2 (en) 1996-10-31 1997-10-30 Streptococcus pneumoniae antigens and vaccines
CA002269663A CA2269663A1 (en) 1996-10-31 1997-10-30 Streptococcus pneumoniae antigens and vaccines
AU51945/98A AU5194598A (en) 1996-10-31 1997-10-30 (streptococcus pneumoniae) antigens and vaccines
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