CN1239514A - 将多肽被囊化于淀粉基质中 - Google Patents
将多肽被囊化于淀粉基质中 Download PDFInfo
- Publication number
- CN1239514A CN1239514A CN97180232A CN97180232A CN1239514A CN 1239514 A CN1239514 A CN 1239514A CN 97180232 A CN97180232 A CN 97180232A CN 97180232 A CN97180232 A CN 97180232A CN 1239514 A CN1239514 A CN 1239514A
- Authority
- CN
- China
- Prior art keywords
- gly
- ala
- val
- leu
- asp
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
- C12N15/8251—Amino acid content, e.g. synthetic storage proteins, altering amino acid biosynthesis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
- C12N15/8245—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Abstract
提供了杂合多肽,该多肽是与源于编码合成代谢蛋白的基因的被囊化部分一起生成的。更具体地讲,本发明涉及编码将结合的蛋白被囊化于一种基质的基因的重组核酸分子;优选地,所述基因把一种(“有效负荷”)多肽被囊化于淀粉中,更具体地讲,被囊化于淀粉粒基质中。还提供了含有所述重组核酸分子的表达载体及其宿主,尤其是由所述载体转化的所述宿主的含有淀粉的部分。优选地,提供了含有被囊化于淀粉中的外源蛋白的谷物,所述谷物可用于生产哺乳类、鱼类、禽类食品。本发明还涉及从淀粉中,尤其是从淀粉中纯化蛋白的方法以及这种蛋白的工业用途。
Description
本申请要求申请日为1996年9月30日的临时专利申请流水号为60/026,855的优先权。所述临时申请以与本申请不一致的程度收作本文的参考文献。
本发明背景多糖酶
原核和真核细胞均用多糖酶作为储存物质。在原核细胞中,原始储存多糖是糖原。尽管糖原与存在于大部分维管植物中的淀粉类似,但它们在链长和聚合度方面存在差异。在许多植物中,淀粉被用作初始的储存多糖。淀粉被储存在含有淀粉的植物的各种组织中。在大多数情况下,淀粉是由两种成分组成:一种是直链淀粉,另一种是支链淀粉。直链淀粉形成线性葡聚糖,而支链淀粉形成葡聚糖的支链。典型的淀粉具有比例为25%的直链淀粉和75%的支链淀粉。植物中直链淀粉与支链淀粉的比率变化会影响其淀粉的性质。此外,来自不同植物的淀粉通常具有不同的性质。玉米淀粉和马铃薯淀粉由于磷酸基团的有或无而有所不同。某些植物的淀粉性质因为引入该植物基因组中的突变而不同。玉米、稻和豌豆等里的突变型淀粉是众所周知的。
淀粉分支或淀粉成分比例的改变会产生不同的淀粉特性。淀粉的一种特性是生成淀粉粒,淀粉粒特别生成于叶、根、块茎和种子中。这些淀粉粒是在淀粉合成过程中形成的。某些淀粉合成酶特别是结合在颗粒中的淀粉合成酶、可溶性淀粉合成酶和分支酶是在淀粉粒形成时“被囊化”于淀粉粒中的蛋白。
在国际专利申请公开号GB92/01881中披露了动物和细菌糖原合成酶cDNA克隆的用途。从该文献中可以了解糖原合成酶的核苷酸序列和氨基酸序列。例如,可以从存储号为J02616(Kumar等,1986,生物化学杂志,261:16256-16259)的GenBank/EMBL(SWISSPROT)数据库中获取编码糖原合成酶的大肠杆菌(E.Coli)glgA基因的核苷酸序列.Okita等(1981,生物化学杂志,256(13):6944-6952)还克隆了大肠杆菌糖原生物合成酶结构基因。所述糖原合成酶glgA结构基因是由Leung等(1987,细菌学杂志169(9)4349-4354)从鼠伤寒沙门氏菌(Salmonellatyphimurium)LT2中克隆的。源于兔骨骼肌(Zhang等,1989,FASEB杂志,3:2532-2536)和人肌肉(Browner等,1989,Proc.Natl.Acad.sci.,86:1443-1447)的糖原合成酶序列也是已知的。
业已报导过植物可溶性淀粉合成酶cDNA克隆的用途。Dry等(1991,植物杂志,2:193-202)披露了豌豆可溶性淀粉合成酶的同种型I和II氨基酸序列。稻可溶性淀粉合成酶的氨基酸序列由Baba等披露(1993,植物生理学)。所述后一种序列(稻SSTS)错误地引用了其N-末端序列,因此是误导。推测这是因为与蛋白酶降解或所提取酶其他固有的不稳定性相关的某些提取失误所致。正确的N-末端序列(始于AESLR)存在于被他们称为稻SSTS转运肽序列中。
Baba等(1991,BBRC,181:8794)研究了玉米分支酶I的序列。Fisher和Shrable(1993,植物生理学,102:1045-1046)研究了源于玉米胚乳的淀粉分支酶II。业已报导了植物、细菌和动物分支酶cDNA克隆的用途。也可从文献中了解细菌分支酶(BE)的核苷酸和氨基酸序列。例如,Kiel等克隆了源于Cyanobacterium synechococusspPPC7942(1989,基因(Amst),78(1)918)和源于嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)(Kiel等,1991,分子基因遗传学,230(12)136-144)的分支酶基因glgB。酿酒酵母(S.cerevisiae)的glc3和gha1基因是等位的并编码所述糖原分支酶(Rowen等,1992,分子细胞生物学12(1)22-29)。Matsumomoto等研究了源于粗糙脉孢酶(Neurospora crassa)的糖原分支酶(1990,生物化学杂志,107:118-122)。所述GenBank/EMBL数据库还含有编码分支酶的大肠杆菌glgB基因的序列。
淀粉分支酶(2.4.1.11)能延长淀粉分子,并被认为可作用于直链淀粉酶和支链淀粉酶。可以证实淀粉合成酶(STS)活性与质体基质中的颗粒相关。与结合淀粉合成酶有关的淀粉生成能力也是公知的。正如Mu-Forster等所披露的(1996,植物生理学111:821-829)现已了解与淀粉生物合成有关的各种酶具有不同的结合能力。颗粒结合淀粉合成酶(GBSTS)活性与waxy基因产物密切相关(Shure等,1983,细胞35:225-233)。业已证实在诸如玉米、稻和马铃薯的多种物种中直链淀粉的合成取决于该基因的表达(Tsai,1974,生化遗传学11:83-96:Hovenkamp-Hermelink等,1987,理论应用遗传学75:217-221)。Visser等披露了源于马铃薯的编码颗粒-结合淀粉合成酶基因的分子克隆和部分鉴定(1989,植物科学64(2):185-192)。Visser等还披露了通过反义构建体抑制马铃薯中编码颗粒-结合淀粉合成酶基因的表达(1991,分子基因遗传学225(2):289-296)。
通过Frydman和Cardini(Frydman和Cardini,1964,生物化学生物物理学研究通讯17:407-411)的开拓性工作,已了解到其他STS酶是可溶性淀粉合成酶。最近,由于发现了所述酶与所述颗粒相关并存在于可溶相中,所述“可溶性”一词的合理性已值得怀疑(Denyer等,1993,植物杂志4:191-198;Denyer等,1995,植物97:57-62;Mu-Forster等,植物生理学111:821-829)。一般的看法是,支链淀粉的生物合成与可溶性淀粉合成酶和淀粉分支酶的相互作用相关。业已在豌豆(Denyer和Smith,1992,植物186:609-617;Dry等,植物杂志,2:193-202)、马铃薯(Edwards等,1995,植物生理学112:89-97;Marshall等,1996,植物细胞8:1121-1135)和稻(Baba等,1993,植物生理学103:565-573)中鉴定和克隆到了可溶性淀粉合成酶的不同同种型,而大麦似乎含有多种同种型,其中的有些与淀粉分支酶有关(Tyynela和Schulman,1994,植物生理学103:565-573)。STS克隆的共同特征是存在着KXGGLGDV共有序列,该序列被认为是所述酶的ADP-Glc结合位点(FURUKAWA等,1990,生物化学杂志265:2086-2090;Furukawa等,1993,生物化学杂志268:23837-23842)。
在玉米中,业已鉴定了被称为同种型I和II的STS的两种可溶性形式(Macdonald和Preiss,1983,植物生理学73:175-178;Boyer和Preiss,1978,糖类研究61:321-334;Pollock和Preiss,1980,生物化学生物物理学204:578-588;Macdonald和Preiss,1985,植物生理学78:849-852:Dang和Boyer,1988,植物化学27:1255-1259;Mu等,1994,植物杂志6:151-159),但这两种形式均未被克隆。最近,已将玉米胚乳STSI活性与存在于可溶性和颗粒相关的部分中的76-kDa多肽关联起来(Mu等,1994,植物杂志6:151-159)。STSII的多肽特征尚属未知。STSI和II具有不同的酶学特性。STSI具有独立于引物的活性,而STSII需要糖原引物来催化葡糖基转移。业已报导过可溶性淀粉合成酶对淀粉沉积具有高的流量控制系数(Jenner等,1993,奥地利植物生理学杂志22:703-709;Keeling等1993,植物191:342-348)并在高温下具有异常的动力学特性(Keeling等1995,奥地利植物生理学杂志21:807-827)。玉米中相应的同种型在最佳温度和稳定性方面具有显著差异。
植物淀粉合成酶(和大肠杆菌糖原合成酶)序列包括序列KTGGL,该序列已知为ADPG结合域。编码任何上述淀粉合成酶蛋白的基因均可用于本发明结构中。
分支酶[α1,4D葡聚糖:α1,4D葡聚糖6D(α1,4D葡聚糖)转移酶(E.C.2.4.1.18)],有时又被称为Q-酶,它能将淀粉酶转化成支链淀粉酶。α1,4D葡聚糖链的一个片段被转移到一个类似葡聚糖链的伯位羟基上。
业已报导过细菌和植物分支酶基因序列(稻胚乳:Nakamura等,1992,植物生理学84:329-335;Nakamura和Yamanouchi,1992,植物生理学99:1256-1266;豌豆:Smith,1988,植物,175:270-279;Bhattcharyya,1989,细胞生物化学杂志,增刊13D:331;玉米胚乳:Singh和Priess,1985,植物生理学79:34-40;VosScherperkeuter等,1989,植物生理学90:75-84;马铃薯:Kossmann等,1991,分子基因遗传学,230(12):39-44;木薯:Salehuzzaman和Visser,1992,植物分子生物学20:809-819)。
在多糖酶领域,存在着利用各种植物物种中的多种淀粉合成酶基因对植物淀粉支路进行工程修饰的载体的报导。其中,与纤维素或淀粉或糖原结合的上述多糖酶中的一些是众所周知的。利用多糖酶的一个具体的专利实例证明了糖原生物合成酶在修饰植物淀粉方面的用途。在授予Shewmaker的美国专利US5,349,123中披露了一种可在植物细胞中生成糖原生物合成酶的含有DNA的载体。具体地讲,该专利涉及因引入上述酶而对马铃薯淀粉造成的改变。还报导过其它淀粉合成基因及其用途。杂合(融合)多肽
杂合蛋白(也被称为“融合蛋白”)是由融合成单一多肽的两种或两种以上蛋白组成的多肽链。通常,所述蛋白之一是能与特定受体细胞结合的配体。编码融合肽的载体主要被用于通过微生物发酵生产外源蛋白。然后可通过亲和层析纯化所产生的融合蛋白。所述多肽之一的结合部分用于把所述杂合多肽连接在一种亲和基质上。例如,能与β-半乳糖苷酶形成融合蛋白,β-半乳糖苷酶可结合于一种柱上。该方法已被用于生产病毒抗原。
另一种用途是回收所述杂合多肽的多肽之一。裂解所述融合肽的化学和生物学方法是众所周知的。如果在所述肽与肽之间采用的是酸不稳定性天冬氨酰-脯氨酸连键,可以用低PH裂解所述肽,而这些肽不受酸的影响。业已用溴化氰裂解过激素。另外,业已报导过通过位点专一性蛋白水解作用进行的裂解。业已用聚精氨酸尾加强了诸如离子层析其它蛋白纯化方法,所述聚精氨酸尾可提高所述蛋白的总的碱性,从而加强与离子交换柱的结合力。
有很多专利已对制备杂合肽或用于特殊目的的特殊杂合肽的方法的改进作过介绍。授予Pastan等的美国专利US5,635,599披露了对杂合蛋白的一种改进。该专利披露了一种作为所述杂合蛋白部分的循环配体。该配体具有专一性和良好的结合力。在授予Kuliopulos的美国专利US5,648,244中披露了对杂合蛋白的另一种改进。该专利披露了一种用一种载体肽生产杂合肽的方法。当该核酸被一种限制性内切酶识别后可产生非回文3-碱基突出端。这样使得所述载体能被裂解。
在美国专利US5,643,756中报道了具有特殊针对性的杂合蛋白的例子。该专利披露了一种用于在细胞中表达糖基化蛋白的载体。该杂合蛋白适用于HIVgp120的正确免疫反应。用所报导的载体可以加强被高度糖基化的gp120结合域的提取。
美国专利US5,202,247和5,137,819讨论了具有多糖结合域的杂合蛋白,以及用于制备能与一种多糖基质结合的杂合蛋白的方法和组合物。美国专利US5,202,247特别介绍了一种将纤维素结合区与感兴趣的肽连接的杂合蛋白。该专利披露所述杂合蛋白可以在细菌宿主中表达以后通过与纤维素的亲和层析而被纯化。
遗传工程技术的发展使得将来自各种生物和植物的基因转移到其它生物和植物中成为可能。尽管过去业已通过转化和诱变对淀粉作过修饰,仍有必要对淀粉作进一步的修饰。为此,需要下列载体,该载体可将希望的氨基酸或肽被囊化于淀粉中,尤其是淀粉粒中。所得到的淀粉是修饰过的,而且源于带有所述载体的植物的组织也是修饰过的。
本发明概述
本发明提供了一种杂合多肽,该多肽包括一个源于一种淀粉结合酶淀粉被囊化区(SER),该区与一种有效负荷(payload)多肽融合,该有效负荷多肽对于所述淀粉被囊化区来说不是内源性的,即并非是天然与所述淀粉被囊化区连接的。可将这种杂合多肽用于制备含有所述有效负荷多肽的改性淀粉。可将这种改性淀粉用于生产富积了某些氨基酸的谷类饲料。这种改性淀粉还可用于生产淀粉被囊化形式的诸如激素和其它药物,如胰岛素的多肽,以便抵抗胃酸的降解作用。还可将所述杂合多肽用于生产易纯化形式的所述有效负荷多肽。例如,通过细菌发酵生产的或由谷类或动物生产的所述杂合多肽可以从所述改性淀粉中提取和纯化,所述杂合多肽是通过已知方法与所述改性淀粉结合的。
在本文中,“多肽”一词是指多个相同或不同的氨基酸,而且,还包括蛋白。
“杂合多肽”一词是指由源于至少两种不同来源的肽或多肽组成的多肽,例如,由一种淀粉结合酶的淀粉被囊化区与诸如激素的另一种多肽融合,其中,该杂合多肽的至少两个组成部分并非是天然融合在一起的。
“有效负荷多肽”一词是指一种对于所述淀粉被囊化区来说不是内源性的多肽,其表达必须与该区相结合,以便表达一种含有所述有效负荷多肽的改性淀粉。
当所述有效负荷多肽被用于提高所述改性淀粉中特定氨基酸的氨基酸含量时,该多肽优选由不超过3种以上的不同类型的氨基酸组成,这些氨基酸选自下列一组:Ala,Arg,Asn,Asp,Cys,Gln,Glu,Gly,His,Ile,Leu,Lys,Met,Phe,Pro,Ser,Thr,Trp,Tyr,和Val。
正如本领域中所公知的,当所述有效负荷多肽被用于向宿主生物或其它生物提供生物活性多肽时,所述有效负荷多肽可以是诸如激素的生物活性多肽,例如,胰岛素、生长因子,如促生长素、抗体、酶、免疫球蛋白或染料,或者是其生物活性片段。只要所述多肽具有生物学活性,它就不必是天然存在的多肽,而且可以是突变的、截短的、或其它修饰形式。所述生物活性多肽可以是仅含有生物活性多肽的生物活性部分的修饰肽。它们还可以是与天然存在的生物活性氨基酸序列同源的氨基酸序列(优选至少约有75%的同源性),该序列仍具有生物学活性。
所述杂合多肽的淀粉被囊化区可以是本领域已知任何淀粉结合酶的被囊化区,例如,选自下列一组的酶:可溶性淀粉酶I、可溶性淀粉酶II、可溶性淀粉酶III、颗粒结合淀粉合成酶、分支酶I、分支酶IIa、分支酶IIBb和葡糖淀粉酶多肽。
当把所述杂合多肽用于生产纯化或部分纯化形式的有效负荷多肽时,所述杂合多肽优选包括一个位于所述淀粉被囊化区和所述有效负荷多肽之间的裂解位点。因此,分离纯化有效负荷多肽的方法包括让所述杂合多肽与一种对所述裂解位点专一的裂解剂结合的步骤。
本发明还提供了编码所述杂合多肽的重组核酸分子(RNA或DNA)。所述重组核酸分子优选包括适于在所选的宿主中表达所述杂合多肽的控制序列。所述“控制序列”一词包括启动子、内含子、所述特定宿主生物的优选密码子序列,以及本领域已知的能影响DNA或RNA在特定宿主中表达的其它序列。编码所述淀粉被囊化区和有效负荷多肽的核酸序列可以是天然存在的核酸序列,或其生物学活性片段,或者是与所述序列同源的生物活性序列,优选与所述序列有大约75%的同源性。
宿主生物包括细菌、植物和动物。优选的宿主是植物。单子叶植物和双子叶植物均为适用于表达本发明杂合多肽的宿主。
本发明还提供了含有编码本发明杂合蛋白的核酸的表达载体。该表达载体可被用于将所述核酸转化入宿主生物内,并且,还可以含有辅助所述核酸在宿主生物中表达的序列。所述表达载体可以是本领域已知的可用于转化系统的质粒,修饰过的病毒,或DNA或RNA分子,或其他载体。
通过本发明方法,可以生产出含有能够表达本发明杂合多肽的重组核酸分子的转化细胞。所述细胞可以是源于单细胞生物、植物或动物的原核或真核细胞。它们可以是细菌细胞,从该细胞中可收获所述杂合多肽。或者,它们可以是能够再生成植株的植物细胞,从所再生的植株中可以收获所述杂合多肽,或者所述植物细胞可以再生成可育的植株,其所结种子含有编码所述杂合多肽的核酸。在一种优选实施方案中,所述种子含有包括所述有效负荷多肽的改性淀粉。所述“改性淀粉”一词是指已被修饰成含有所述有效负荷多肽的天然存在的淀粉。
还提供了一种对有效负荷多肽进行定向消化以生成该消化过程的特定相,例如,防止有效负荷多肽在动物的胃中被降解的方法,该方法包括给所述动物喂饲本发明的含有所述有效负荷多肽的改性淀粉,以便将所述多肽保护起来,防止其在动物的胃中降解。另外,所述淀粉可以是一种已知能在所述胃中被降解的淀粉,以便在那里释放出所述有效负荷多肽。
本发明的优选重组核酸分子包括编码淀粉被囊化区的DNA,该DNA选自在本发明的表中所列出的淀粉合成基因序列。
本发明的优选质粒适用于特定的宿主。本发明提供了含有一个启动子、一个质体定向序列、一个编码一种淀粉被囊化区的核酸序列、和一个中止子序列的质粒。所述质粒适于插入编码有效负荷多肽和淀粉被囊化区的DNA序列,以便在特定宿主中表达。
本发明的质粒可选择性地包括一个间隔单位或一个连接单位,该单位接近编码SER的核酸与编码所述有效负荷多肽的核酸之间的融合位点。本发明包括含有适于原核或真核宿主的启动子的质粒。所述启动子也可以特别适于在单子叶植物或双子叶植物中表达。
一种制备本发明肽改性淀粉的方法包括以下步骤:提供一种质粒,该质粒具有与编码一种淀粉被囊化区的核酸序列结合的启动子,所述编码淀粉被囊化区的核酸序列与编码一种有效负荷多肽的核酸片段结合,并用所述质粒转化宿主,以使该宿主表达肽改性的淀粉。
本发明还包括含有淀粉的谷物,该谷物包括:胚,营养组织;和将一种蛋白被囊化在其中的改性淀粉粒,该蛋白对于未被改性的所述谷物的淀粉粒来说是非内源性的。所述含有淀粉的谷物可以是这样的谷物:其中所述胚是玉米胚、稻胚或小麦胚。
本文所提到的所有参考文献均被以不同程度地收作本文的参考文献。
附图的简要说明
图1a表示质粒pEXS114,该质粒含有被亚克隆到Stratagene出售的pBSK中的合成GFP(绿色荧光蛋白)。
图1b表示质粒pEXS115。
图2a表示具有被亚克隆到一种市售质粒中的限制位点的waxy基因。
图2b表示由Novagen出售的pET-21A质粒,该质粒具有亚克隆在其上面的源于pEXS115的GFP片段。
图3a表示被亚克隆到pEXSWX上的pEXS114,以及GFP-FLWX图谱。
图3b表示GFP-BamHIWX质粒。
图4表示被亚克隆到pEXSWX上的pEXS115的SG6FP片段,以及GFP-NcoWX图谱。
图5表示一种适用于单子叶植物的质粒的线性示意图。
图6表示质粒pEXS52。
图7表示用于制备pEXS51和pEXS60的6个引入质粒。图7a表示pEXS adh1。图7b表示pEXS adh1-nos3’。图7c表示pEXS33。图7d表示pEXS10zp。图7e表示pEXS10zp-adh1。图7 f表示pEXS10zp-adh1-nos3’。
图8a和8b分别表示质粒pEXS50和pEXS51,这些质粒含有MS-SIII基因,该基因是可溶性淀粉合成酶基因。
图9a表示排除了pEXS50中所示内含子的质粒pEXS60,而图9b表示排除了pEXS60中的内含子的质粒pEXS61。
详细说明
广义地讲,本发明提供了一种杂合多肽,一种生产杂合多肽,以及编码所述杂合多肽的核酸的方法。杂合多肽包括被融合成一个单一肽链的两个或两个以上亚单位。所述亚单位可以是氨基酸或肽或多肽。所述亚单位之一是一个淀粉被囊化区。因此,杂合多肽可以被引导至由表达该杂合多肽的生物所产生的淀粉粒中。
一种在细胞中生产杂合多肽的方法包括制备一种DNA构建体,该构建体包括至少一个编码一种起着将所述结合的DNA的表达产物结合到淀粉粒中的作用的序列的片段,并与编码所述感兴趣的多肽(有效负荷多肽)的DNA序列连接。该构建体在真核或原核细胞中表达。可以用所述杂合多肽生产纯化的蛋白或将一种感兴趣的蛋白固定在淀粉粒中使其受到保护,或用于生产含有外源氨基酸或肽的谷物。本发明的杂合多肽具有三个部分。
有效负荷肽(X) | 中央位点(CS)* | 淀粉被囊化区(SER) |
X是任何感兴趣的氨基酸或肽。
*是可选部分。
编码X的基因可被置于下述DNA构建体中的5’或3’位置。
CS是一个中央位点,该位点可以是本领域已知的离去位点、裂解位点或间隔子。裂解位点可以由一种裂解酶识别。裂解酶是能在特定位点裂解肽的酶。被用于裂解多肽的化合物或酶的例子包括凝血酶、胰蛋白酶、溴化氰、甲酸、羟胺、胶原酶、和alasubtilisin。间隔子是一种肽,由它连接构成所述杂合多肽的肽。通常,除了连接所述肽或保持某种最低间距或影响该蛋白的折叠、电荷或水可及性外它不具备任何特定活性。间隔子可以是不影响所述杂合多肽的生物学活性的任何肽序列。
所述淀粉被囊化区(SER)是目标多肽的对淀粉具有结合亲和力的部分。通常,SER选自下列一组肽:包括植物淀粉合成酶和分支酶的淀粉结合区,但可以包括源于诸如葡糖淀粉酶等的其它来源的淀粉结合域。在本发明的优选实施方案中,所述SER包括天然存在于淀粉合成途径上的基因的肽产物。这一小类优选SERs被称为成淀粉被囊化区(SFER)。本发明优选的SERs的另一小类是源于含有淀粉的植物的特殊酶——淀粉合成酶(STS)、颗粒结合淀粉合成酶(GBSTS)和分支酶(BE)的特定淀粉被囊化区(SSER)。这一小类的最优选的基因产物的是GBSTS,另外,淀粉合成酶I和分支酶II是有用的基因产物。优选地,所述SER(即上面所述的所有小类)是全长淀粉合成酶基因的截短形式,使其截短的部分包括所述淀粉被囊化区。
广义地讲,用于在所述宿主中表达所述杂合多肽的DNA构建体如下:
启动子 | 内含子* | 转运肽编码区* | X | SER | 终止子 |
*可选部分。还可以使用其它可选部分。
正如本领域所公知的,启动子是控制转录的DNA区域。为不同的宿主选择不同类型的启动子。Lac启动子和T7启动子适用于原核生物,35ScaMV启动子适用于双子叶植物,而许多聚遍在蛋白启动子适用于许多单子叶植物。本领域已知的多种不同启动子的任一种均可用于本发明范围内。
另外,正如本领域技术人员所公知的,内含子是基因中不编码基因产物的核苷酸序列。通常能提高在单子叶植物中的表达的内含子的质体中形成淀粉粒,但对转运肽编码区是编码蛋白向诸如质体的细胞器中转运的核苷酸序列。优选选用能被在其中使用该转运多肽的宿主识别并与其相容的转运多肽。在本发明中,所选择的质体是造粉体。
优选的是,所述杂合多肽位于诸如植物细胞的细胞中的造粉体中,所述细胞合成淀粉并将其储存在造粉体中。如果所述宿主是不含造粉体的细菌或其它细胞,则无须转运肽编码区。
终止子是终止所述转录的DNA序列。
X是所述有效负荷多肽的编码区,该多肽可以是感兴趣的任何多肽或氨基酸链。它可以具有一种已知多肽的完整序列或含有其有用的片段。所述有效负荷多肽可以是多肽、或其片段、或生物活性蛋白,它是酶、激素、生长因子、免疫球蛋白、染料等。可用于本发明中的所述有效负荷多肽的某些例子包括,但不限于促乳素(PRL)、血清白蛋白、生长因子和生长激素,即促生长素。血清白蛋白包括牛、绵羊、马、禽和人血清白蛋白。生长因子包括表皮生长因子(E6F)、胰岛素样生长因子I(IGFF-I)、胰岛素样生长因子II(IGFF-II)、成纤维细胞生长因子(FGF)、转化生长因子α(TGF-α)、转化生长因子β(TGF-β)、神经生长因子(NGF)、血小板衍生的生长因子(PDGF)、和重组人胰岛素样生长因子I(rHuIGFF-I)和II(rHuIGFF-II)。可用于实施本发明的促生长素包括,但不限于牛、猪、绵羊、马、禽和人促生长素。猪促生长素包括δ-7重组猪促生长素,正如在欧洲专利申请公开号104,920(Biogen)中所披露并要求保护的。优选的有效负荷多肽是促生长素、胰岛素A链和B链、降钙素、β内啡肽、尿抑胃素、β珠蛋白、肌红蛋白、人生长激素、血管紧张肽、脯氨酸、蛋白酶、β-半乳糖苷酶、和纤维素酶。
所述杂合多肽、SER区和有效负荷多肽还可以本领域已知的翻译后修饰,如糖基化、乙酰化和不会影响所述多肽的期望活性的其它修饰。
开发一种杂合多肽
所述SER区存在于与淀粉合成有关的基因中。分离这种基因的方法包括从基因组DNA文库和从cDNA文库中筛选。可以通过连接、诱变剂、消化、限制和其它诸如此类的方法对基因进行裂解和改变,例如,由Maniatis在分子克隆一书(冷泉港实验室,冷泉港,纽约)中所披露的。用于获取所述SER区的优良原材料的例子包括,但不限于以下诸种:淀粉合成酶I、II、III、IV,分支酶I、IIA和B,以及颗粒结合淀粉合成酶(GBSTS)。所述基因存在于诸如稻、玉米、豌豆、马铃薯和小麦等的含有淀粉的植物中。利用由基因组DNA或cDNA或mRNA制备的SER探针或抗SER的抗体可以分离并鉴定到可用于克隆的基因。可以对淀粉酶编码序列进行修饰,只要这种修饰不影响所述SER区对相关多肽进行被囊化的能力即可。
一旦对编码被被囊化到淀粉粒中的蛋白进行定位,然后就可以用本领域已知的几种方法分离所述SER。一种方法是用限制酶在各种位点裂解所述基因,使其N-末端缺失,并让所得到的蛋白表达。然后让所表达的截短了的蛋白在淀粉凝胶上运动,以评价残留蛋白的结合和解离常数。可以将诸如绿色荧光蛋白的本领域已知的标记基因连接在截短了的蛋白上,并用于确定该标记基因在淀粉粒中的存在。
一旦分离到SER基因序列片段,即可将其用于制备能够表达被被囊化于淀粉中的有效负荷多肽的基因片段序列。可将SER基因序列和编码所述有效负荷多肽的基因序列连接在一起。然后,就可以将融合的DNA置于多种载体结构中,以便在多种宿主中表达。优选的宿主在质体中形成淀粉粒但SER的试验可以在诸如大肠杆菌的细菌性宿主中方便地进行。
编码所述有效负荷多肽的核酸序列可由DNA、RNA、基因组DNA、eDNA、mRNA衍生而来,或者也可以是全面或部分合成。可以操作所述有效负荷多肽,使其含有突变,使所产生的蛋白是一种新的突变蛋白,只要保留了生物学功能即可。
当所述有效负荷多肽编码核酸序列被连接到SER编码序列上后,编码有效负荷多肽的基因序列优选结合在编码其N-末端的SER序列的末端。尽管所述N-末端是优选的,但对于本发明来说,所述有效负荷多肽是连接于SERN-末端或是C-末端似乎并不重要。很明显,制备本发明重组核酸分子的方法,无论是合成或是通过克隆和连接进行的,对于本发明来说并不重要。
所述杂合多肽的中央部分是选择性的。对于本发明的某些应用来说,将位于该部分的编码一种常见蛋白酶裂解位点的DNA引入用于表达所述杂合多肽的重组核酸分子中可能是十分有用的。另外,将编码对pH敏感的氨基酸序列的DNA引入以形成所述中央部分也是很有用的。如果将本发明用于开发一种能够用淀粉酶等提取并从淀粉粒中释出的纯化蛋白,则可以利用淀粉酶裂解位点。另外,如果是在动物体内消化所述蛋白,则蛋白酶裂解位点有助于动物消化道内的酶从淀粉中释放出该蛋白。在其它用途中,而且在很多消化用途中,所述裂解位点是多余的。
所述中央部分位点可以包括一个间隔子。间隔子是指连接构成杂合多肽的蛋白的肽。一般,除了连接所述蛋白、保持某种最低间距、影响该杂合多肽的折叠、电荷或疏水或亲水性质外,该间隔子不具有任何特殊功能。
构建体形成
一旦生成编码所述杂合多肽的连接DNA,即可制备出克隆载体或质粒,这种载体或质粒能将所述DNA转入一种宿主,以表达所述杂合多肽。将本发明的重组核酸序列插入常见克隆载体或质粒中。对于本发明来说,优选宿主是产淀粉粒宿主。不过,也可以使用细菌宿主。尤其有用的是被转化成含有一种植物的部分或全部淀粉合成基因的细菌宿主。本领域普通技术人员了解,所述质粒适用于所述宿主。例如,在细菌宿主中,转录调节启动子包括lac、TAC、和trp等。另外,最好不使用编码转运肽的DNA,并且可以用位于结构基因上游的分泌前导序列将所述多肽分泌入培养基中。或者,所述产物任留在宿主中,裂解该宿主,并通过淀粉提取方法分离和纯化该产物,或通过让该材料与淀粉基质(或诸如直链淀粉或支链淀粉糖原等的淀粉样基质)结合,以提取所述产物。
优选的宿主是植物,因此,优选的质粒适用于植物中。该质粒应含有一个启动子,优选为适于在该植物的含淀粉组织中表达的启动子。该启动子可以对诸如种子、根、和块茎之类的各种组织有专一性;或者,也可以是在植物组织中进行基因表达的组成型启动子。众所周知的启动子包括10kD玉米醇溶蛋白(玉米)启动子、CAB启动子、patastin、35S和19S花椰菜花叶病毒启动子(在双子叶植物中十分有用)、聚遍在蛋白启动子(可用于单子叶植物)、以及本领域已知的其增强或改进形式。
所述克隆载体可以含有编码一种转运肽的序列,由该转运肽引导质粒进入正确位置。可以使用其它转运肽的编码序列。优选使用天然存在于宿主中的转运肽。玉米的优选转运肽编码区示于本文的表和图中。该转运肽的用途是引导载体进入正确的胞内区。
编码所述有效负荷多肽的N-末端的DNA序列与所述转运肽编码序列结合。编码有效负荷多肽的序列的方向,根据是否需要正义或反义转录而变化。本文所披露的本发明的DNA构建体,其编码所述有效负荷多肽的序列位于N-末端,不过,SER编码区也可以位于其N-末端,而所述有效负荷多肽位于其后。在该DNA构建体末端是终止子序列。该序列是本领域中公知的。
将所述克隆载体转入一种宿主。可以用本领域已知的多种转化技术将所述克隆载体(优选为质粒)导入宿主中。所述技术可以因宿主而改变,不过,这类技术包括微粒子轰击、微量注射、农杆菌转化、“whiskers”技术(US专利5,302,523和5,464,765)、和电击法等。如果所述宿主是植物,可将其细胞再生成植株。再生植株的方法为本领域所公知。所述宿主一旦被转化,所述蛋白在其中得到表达,即可证实编码所述有效负荷多肽的DNA的存在。可以通过Western印迹或EKISA或根据植株或细胞中改变的结果证实表达蛋白的存在。
被囊化蛋白的用途
本发明具有多种用途。可以纯化形式从所述淀粉上裂解所述杂合多肽(可以包括裂解位点),并可以回收纯化蛋白。另外,还可以原始形式使用存在于淀粉中的所述被囊化有效负荷多肽,将蛋白输送到食用动物消化道的各个部分(“动物”包括哺乳动物、禽类和鱼)例如,如果将所述材料被囊化在其中的淀粉能承受消化,则蛋白会缓慢释入动物的肠中,从而避免有价值的蛋白在其胃中降解。可将诸如甲硫氨酸和赖氨酸的氨基酸被囊化,直接掺入饲养动物的谷物中,从而消除了以其它形式向其饲料中添加这些氨基酸的必要性。
发明还可以把激素、酶、蛋白、蛋白类养分、和蛋白类药物输送到动物消化道的特定消化区。蛋白通常是在上消化道被消化,将其被囊化在淀粉中能使其以非消化形式通过胃,并由其肠道完全或部分吸收。如果能通过其肠壁,即可将所述有效负荷多肽用于治疗动物,或提供诸如生长因子的激素,如促生长素,以便对动物进行免疫接种,或加强给动物的营养。
如果所述淀粉在胃中不能承受消化作用(例如,含糖的2型淀粉是高度可消化的),则可以让所添加的蛋白由所述动物的上消化道定向吸收。这就需要对产生所述改性淀粉的宿主进行突变或转化,使其能产生含糖的2型淀粉。本发明包括将能产生改性淀粉的突变型生物用作宿主。此类突变型宿主的某些例子包括具有含糖的1型、含糖的2型、脆型、皱缩型、蜡质型、直链淀粉延长型、暗色型、不透明型、和粉质型突变等的稻和玉米等。所述突变型淀粉和源于不同植物的淀粉具有不同程度的可消化性。因此,通过选择用于表达所述DNA的宿主和喂饲所述改性淀粉的动物,可以在指定部位消化所述杂合多肽。不同的蛋白能被身体的不同部位最有效的吸收。通过将所述蛋白被囊化于具有特定可消化性的淀粉中,可以在消化过程的特定时间,将所述蛋白输送到其消化道的任何部位。
本发明的另一个优点是,抑制或表达不同水平所期望的多肽的糖基化作用的能力。所述被囊化过程能使所述蛋白在淀粉粒中表达时的糖基化状态不同于由其它DNA分子表达时的糖基化状态。所述糖基化作用取决于被囊化量、所采用的宿主以及所述多肽的序列。
通过对已知其具有其它有利性状点的植物进行遗传操作,可以产生具有上述特征的改进作物。通过操作一种淀粉合成酶基因的核苷酸序列,可以改变在一种植物中所产生的重要氨基酸、蛋白或肽的量。可以通过有性杂交或转化将一个或几个遗传工程基因构建体整合到植物基因组中,所述基因构建体可源于植物、真菌、细菌或动物。工程基因可以含有额外拷贝的野生型基因,或者,可以编码具有新的特性的修饰的或等位的或改型的酶。所述基因结构的整合,根据所引入(沿有义或反义方向)基因的数量和类型而具有不同作用。它可以加强植物产生特殊蛋白、肽的能力或改善氨基酸平衡。
克隆参与淀粉生物合成的酶
可以用已知的克隆技术提供本发明的DNA结构。特定形式SSTS、GBSTS、BE、糖原合成酶(GS)、支链淀粉、或本发明所采用的其它基因的来源可以是能产生淀粉或糖原的任何生物。要对潜在的供体生物进行筛选和鉴定。然后可以采用以下两种方法:(a)在本文所披露的方法之后采用酶纯化和抗体/序列生成方法;(b)用SSTS、GBSTS、BE、糖原合成酶(GS)、支链淀粉、或其它cDNAs作为异源探针,在源于相关生物的文库中鉴定编码SSTS、GBSTS、BE、糖原合成酶(GS)、支链淀粉、或其它淀粉被囊化酶的基因组DNAs。基因转化、植株再生和检测方法是本领域公知的。在这种场合,有必要使用于转化的基因结构含有调控序列,以确保其在淀粉生成期间表达。这种调控序列存在于小型谷物和块茎及根中。例如,这种调控序列常见于玉米胚乳的编码颗粒结合淀粉合成酶(GBSTS)、可溶性淀粉合成酶(SSTS)或分支酶(BE)或其它玉米胚乳淀粉合成途径酶的DNA上。这些源于所述胚乳的调控序列可确保蛋白在正确的发育时间表达(例如,ADPG焦磷酸酶)。
在该方法中,我们在有适当浓度的诸如糖原或支链淀粉的糖类的存在下,用天然蛋白电泳的方法测定淀粉结合蛋白的淀粉结合常数。用本领域技术人员熟知的定点突变和其他遗传工程方法,能说明淀粉囊化区。可以用本文所披露的方法评价带有新的肽或氨基酸组合的新型遗传工程蛋白。
实施例
例1:
鉴定淀粉被囊化蛋白的方法
淀粉-颗粒蛋白分离:
在Waring搅拌器中用25ml提取缓冲液(50mMTris乙酸,pH7.5,1mMEDTA,1mMDTT)对12.5g谷物进行3×20秒的匀浆,每次匀浆间隔1分钟。将样品保持在冰上。通过纱布过滤,并在6,000rpm下离心30分钟。弃上清液,并刮去覆盖白色淀粉沉淀的脱色固体。将沉淀再悬浮于25m1缓冲液中,并再次离心。再重复洗涤两次。将洗涤过的沉淀再悬浮于-20℃丙酮中,让沉淀在-20℃下沉降。重复。在空气流下干燥淀粉。在-20℃下贮存。
蛋白提取
将50mg淀粉与1ml2%SDS混合于微量离心管中。涡旋混合,在4℃下以18,000rpm的速度离心5分钟。倒掉上清夜。重复2次。加入1ml样品缓冲液(4ml蒸馏水,1ml 0.5MTris-HCL,pH6.8,0.8ml甘油,1.6ml10%SDS,0.4mlB-巯基乙醇,O.2ml0.5%溴酚蓝)。将微量离心管煮沸10分钟,在盖上开孔。冷却,10,000rpm下离心10分钟。将上清液倾入新的微量离心管中。用标准方法煮沸4分钟。冷却。
SDS-PAGE凝胶:(未变性)
10%分离胶 4%堆积胶
Acryl/Bis40%母液 2.5ml 1.0ml
1.5M Tris pH8.8 2.5ml -
0.5M Tris pH8.8 - 2.5ml
10%SDS 100μl 100μl
水 4.845ml 6.34ml
脱气15分钟,加入
新的10%过硫酸铵 50μl 50μl
TEMED 5μl 10μl
Mini-ProteanII Dual Slab小室;每块凝胶用3.5ml分离缓冲液。将4%的堆积胶倾在上部。在200V的恒定电压下对所述凝胶进行电泳。电泳是在10x电泳缓冲液(250mMTris,1.92M甘油,1%SDS,pH8.3)中进行的。
测定淀粉被囊化区的方法:
溶液:
提取缓冲液: 50mMTris乙酸,pH7.5,10mMEDTA,10%蔗
糖,2.5mM新配制的DTT。
堆积缓冲液: 0.5MTris-HCL,pH6.8
分离缓冲液: 1.5MTris-HCL,pH6.8
10X下电极缓冲液: 30.3gTris+144g甘氨酸,将体积调至1L(pH
为大约8.3,不作调整)。稀释后使用。
上电极缓冲液: 同下电极缓冲液
蔗糖溶液: 18.66g蔗糖+100ml蒸馏水
30%Acryl/Bis母液 146g丙烯酰胺+4gbis+350ml蒸馏水
(2.67%C): 调至500ml。过滤,并在4℃下于黑暗中保
存达1个月。
15%Acryl/Bis母液 6g丙烯酰胺+1.5gbis+25ml蒸馏水
(20%C): 调至50ml。过滤,并在4℃下于黑暗中保存
达1个月。
核黄素溶液: 1.4g核黄素+100ml蒸馏水。于黑暗中保
存达1个月。
SS测定混合物: 25mM柠檬酸钠,25mMBicine-NaOH(pH8.O),
2mMEDTA,1mM新配制的DTT,1mM新配制
的腺苷5’二磷酸葡萄糖,10mg/ml新配制
的III型兔肝糖原。
碘溶液: 2g碘+20gKI,加0.1N HCl至1L。
提取:
· 4ml提取缓冲液+12g胚乳。匀浆。
· 通过米拉布或4层纱布过滤,20,000g(14,500rpm,SM-24
转子)离心,
20分钟,4℃。
· 用玻璃移液管除去上清液。
· 0.85ml提取液+0.1ml甘油+0.05ml0.5%溴酚蓝。
· 涡旋并全速离心5分钟。直接使用或冷冻于液氮中并在-80
℃下保存长达2周。
浇灌凝胶:
用透明胶带将Gel Bond PAG膜(FMC Industries,Rockland,ME)连接在外玻板(的内侧)上,使亲水一侧向上。调节所述胶带和膜,使其尽可能紧密和均匀地接触所述玻板的底部。所述膜略小于玻板。将水注入所述膜和玻板之间,以吸附该膜。用织物挤出多余的水。用常用方法组装玻板,然后用粘性粘合剂密封所述玻板的底部。如果已将格雷氏橡胶(gray rubber)从所述浇灌架上取出,即可将所述组装件放入该浇灌架中。所述凝胶与所述膜聚合,并在随后的所有操作中保持接触。
浇灌4.5%T分离小凝胶(O.75mm):
2.25ml蒸馏水
+3.75ml蔗糖溶液
+2.5ml分离缓冲液
+1.5m130%Aeryl/Bis母液
+每种凝胶加不同量的糖原(即:O-1.0%)
脱气15分钟
+50μl 10%APS
+5μlTEMED
聚合30分钟或过夜
浇灌3.125%T母液:
1.59ml蒸馏水
+3.75ml蔗糖溶液
+2.5ml堆积缓冲液
+2.083ml 15%Aeryl/Bis母液
不进行脱气
15μl10%APS
+35μl核黄素溶液
+30μlTEMED
靠近一盏灯泡聚合2.5小时
在拔出梳子之前于4℃下冷却。也可以不用梳子,仅浇灌1厘米厚的堆积胶。
上述方法:· 可以在不同温度下进行;预温育凝胶和溶液。· 在200V电压下预电泳15分钟。· 凝胶加样:每孔7μl,如果无梳子的话每孔115μl。· 在140V电压下电泳,直到染料前部接近底部。通过将整个凝胶装
置放在水浴中可以达到各种电泳温度。可以偶尔终止电泳以便将
温度探测头插入凝胶。· 酶测定:切除染料前部的凝胶。在SS中温育。在室温下轻摇测定
混合物过夜。用水漂洗凝胶。用I2/KI溶液冲洗。· 在灯箱上给所述凝胶照相,并测定照片。Rm=从凝胶上部到活性带
处的距离mm/从凝胶上部到其被切割部位(染料前部所在处)底
部的距离。用%糖原量对1/Rm作图。其曲线与x轴的交点是-K(此
时y=0)。
SER片段长度的测定和评价方法:
在上述方法之后,为了选择SER片段需要4个基本步骤。首先,必须选择编码一种具有淀粉被囊化区的蛋白的DNA。例如,可以从诸如淀粉酶基因的已知淀粉合成基因或淀粉结合基因中选择所述DNA。必须提取所述蛋白。有多种蛋白提取技术为本领域所熟知。可以用蛋白酶处理所述蛋白,以生成不同长度的蛋白片段。优选的片段主要是缺失了该蛋白的N-末端片段。与N-末端相比,SER区更靠近C-末端。让所述蛋白在上述凝胶上电泳,并评价对该凝胶基质的亲和力。更高的亲和力表示上述蛋白的所述区域对该基质有优先选择性。该方法可以对不同蛋白进行比较,以鉴定天然或合成蛋白中的淀粉被囊化区。
实施例2:
SER融合载体:
以下的融合载体适用于大肠杆菌中。与该载体上的可检测SER结合的融合基因编码绿色荧光蛋白(GFP)。可将任意数量的编码蛋白和多肽的不同基因连接到该载体上。所构建的一种融合载体具有与第二个基因或基因片段融合的蜡质玉米的SER,在这里,所述第二个基因是GFP。
pEXS114(见图1a):用引物EXS73(5’-GACTAGTCATATG GTG AGCAAG GGC GAG GAG-3’)[序列1]和EXS74(5’-CTAGATCTTCATATG CTTGTA CAG CTC GTC CAT GCC-3’)[序列2],由质粒HBT-SGFP(由分子生物学系的Jen Sheen提供;Wellman 11,MGH;波斯登,MA02114)PCR扩增合成的GFP(SGFP)。用T DNA聚合酶将PCR产物末端补齐,以产生平端;然后用SpeI消化该PCR产物。将该SGFP片段亚克隆到pBSK(Stratagene,11011North Torrey Pines路,La Jolla,加拿大)的EcoRV-Spe I位点,以产生pEXS114。
pEXS115(见图1b):用引物EXS73(同上)和EXS75(5’-CTAGATCTTGGCCATGGC CTT GTA CAG CTC GTC CAT GCC-3’)[序列3],由质粒HBT-SGFP(由Jen Sheen提供)PCR扩增合成的GFP(SGFP)。用T DNA聚合酶将PCR产物末端补齐,以产生平端;然后用SpeI消化该PCR产物。将该SGFP片段亚克隆到pBSK(Stratagene)的EcoRV-SpeI位点,以产生pEXS115。
pEXSWX(见图2a):将玉米WX的NdeI-NotI片段亚克隆到pET-21a上(见图2b)。基因组DNA序列及可由其产生mRNA序列的有关氨基酸示于下面的表1a和1b中,另外,还可以使用列于下表中所列的DNA。
表1a
玉米waxy基因的DNA序列和推定的氨基酸序列
[序列4和序列5]基因座 ZMWAXY 4800bp DNA PLN定义 Zea mays waxy (wx+) locus for UDP-glucose starch glycosyl
transferase.编号 X03935 M24258关键词 葡糖基转移酶 转运肽UDP萄糖淀粉糖基转移酶
waxy基因座来源 玉米生物体 Zea mays
Eukaryota;Plantae;Embryobionta;Magnoliophyta;Liliopsida;
Commelinidae;Cyperales;Poaceae.参考文献 1(bases 1 to 4800)
作者 Kloesgen,R.B.,Gierl,A.,Schwarz-Sommer,Z.and Saedler,H.
题目 Molecular analysis of the waxy locus of Zea mays
杂志 Mol.Gen.Genet.203,237-244(1986)
标准 full automatic注释 NCBI gi:22509特征 Location/Qualifiers
来源 1..4800
/organism="Zea mays"
重复区 283..287
/note="direct repeat 1"
重复区 288..292
/note="direct repeat 1"
重复区
293..297
/note="direct repeat 1"
重复区
298..302
/note="direct repeat 1"
misc feature 372..385
/note="GC stretch(pot.regulatory factor bindingsite)"
misc feature 442..468
/note="GC stretch(pot.regulatory factor bindingsite)"
misc feature 768..782
/note="GC stretch(pot.regulatory factor bindingsire)"
misc feature 810..822
/note="GC stretch (pot.regulatory factor bindingsite)"
misc_feature 821..828
/note="target duplication s
CAAT signal 821..828
TATA signal 867..873
misc feature 887..900
/note="GC stretch(pot.regulatory factor bindingsite)"
misc feature 901
/note="transcriptional start site"
exon 901..1080
/number=1
内含子 1081..1219
/number=1
外显子 1220..1553
/number2
transit peptide 1233..1448
CDS join(1449..1553,1685..1765,1860..1958,2055..2144,2226..2289,2413..2513,2651..2760,2858..3101,3212..3394,
3490..3681,3793..3879,3977..4105,4227..4343)
/note="NCBI gi:22510"
/codon_start=1
/product="glucosyl transferase"/translation="ASAGMNVVFVGAEMAPWSKTGGLGDVLGGLPPAMAANGHRVMVVSPRYDQYKDAWDTSVVSEIKMGDGYETVRFFHCYKRGVDRVFVDHPLFLERVWGKTEEKIYGPVAGTDYRDNQLRFSLLCQAALEAPRILSLNNNPYFSGPYGEDVVFVCNDWHTGPLSCYLKSNYQSHGIYRDAKTAFCIHNISYQGRFAFSDYPELNLPERFKSSFDFIDGYEKPVEGRKINWMKAGILEADRVLTVSPYYAEELISGIARGCELDNIMRLTGITGIVNGMDVSEWDPSRDKYIAVKYDVSTAVEAKALNKEALQAEVGLPVDRNIPLVAFIGRLEEQKGPDVMAAAIPQLMEMVEDVQIVLLGTGKKKFERMLMSAEEKFPGKVRAVVKFNAALAHHIMAGADVLAVTSREEPCGLIQLQGMRYGTPCACASTGGLVDTIIEGKTGFHMGRLSVDCNVVEPADVKKVATTLQRAIKVVGTPAYEEMVRNCMIQDLSWKGPAKNWENVLLSL
GVAGGEPGVEGEEIAPLAKENVAAP"
内含子 1554..1684
/number=2
外显子 1685..1765
/number=3
内含子 1766..1859
/number=3
外显子 1860..1958
/number=4
内含子 1959..2054
/number=4
外显子 2055..2144
/number=5
内含子 2145..2225
/number=5
外显子 2226..2289
/number;6
内含子 2290..2412
/number=6
外显子 2413..2513
/number=7
内含子 2514..2650
/number=7
外显子 2651..2760
/number=8
内含子 2761..2857
/number=8
外显子 2858..3101
/number=9
内含子 3102..3211
/number=9
外显子 3212..3394
/number=10
misc_feature 3358..3365
/note="target duplication site (Ac9)"
内含子 3395..3489
/number=10
外显子 3490..3681
/number=11
misc_feature 3570..3572
/note="target duplication site(Spm 18)"
内含子 3682..3792
/number=11
外显子 3793..3879
/number=12
内含子 3880..3976
/number=12
外显子 3977..4105
/number=13
内含子 4106..4226
/number=13
外显子 4227..4595
/number=14
polyA_signal 4570..4575
polyA_signal 4593..4598
polyA_site 4595
polyA_signal 4597..4602
polyA_site 4618
polyA_site 4625BASE COUNT 935A 1413C 1447G 1005TORIGIN
1 CAGCGACCTA TTACACAGCC CGCTCGGGCC CGCGACGTCG GGACACATCT TCTTCCCCCT
61 TTTGGTGAAG CTCTGCTCGC AGCTGTCCGG CTCCTTGGAC GTTCGTGTGG CAGATTCATC
121 TGTTGTCTCG TCTCCTGTGC TTCCTGGGTA GCTTGTGTAG TGGAGCTGAC ATGGTCTGAG
181 CAGGCTTAAA ATTTGCTCGT AGACGAGGAG TACCAGCACA GCACGTTGCG GATTTCTCTG
241 CCTGTGAAGT GCAACGTCTA GGATTGTCAC ACGCCTTGGT CGCGTCGCGT CGCGTCGCGT
301 CGATGCGGTG GTGAGCAGAG CAGCAACAGC TGGGCGGCCC AACGTTGGCT TCCGTGTCTT
361 CGTCGTACGT ACGCGCGCGC CGGGGACACG CAGCAGAGAG CGGAGAGCGA GCCGTGCACG
421 GGGAGGTGGT GTGGAAGTGG AGCCGCGCGC CCGGCCGCCC GCGCCCGGTG GGCAACCCAA
481 AAGTACCCAC GACAAGCGAA GGCGCCAAAG CGATCCAAGC TCCGGAACGC AACAGCATGC
541 GTCGCGTCGG AGAGCCAGCC ACAAGCAGCC GAGAACCGAA CCGGTGGGCG ACGCGTCATG
601 GGACGGACGC GGGCGACGCT TCCAAACGGG CCACGTACGC CGGCGTGTGC GTGCGTGCAG
661 ACGACAAGCC AAGGCGAGGC AGCCCCCGAT CGGGAAAGCG TTTTGGGCGC GAGCGCTGGC
721 GTGCGGGTCA GTCGCTGGTG CGCAGTGCCG GGGGGAACGG GTATCGTGGG GGGCGCGGGC
781 GGAGGAGAGC GTGGCGAGGG CCGAGAGCAG CGCGCGGCCG GGTCACGCAA CGCGCCCCAC
841 GTACTGCCCT CCCCCTCCGC GCGCGCTAGA AATACCGAGG CCTGGACCGG GGGGGGGCCC
901 CGTCACATCC ATCCATCGAC CGATCGATCG CCACAGCCAA CACCACCCGC CGAGGCGACG
961 CGACAGCCGC CAGGAGGAAG GAATAAACTC ACTGCCAGCC AGTGAAGGGG GAGAAGTGTA
1021 CTGCTCCGTC GACCAGTGCG CGCACCGCCC GGCAGGGCTG CTCATCTCGT CGACGACCAG
1081 GTTCTGTTCC GTTCCGATCC GATCCGATCC TGTCCTTGAG TTTCGTCCAG ATCCTGGCGC
1141 GTATCTGCGT GTTTGATGAT CCAGGTTCTT CGAACCTAAA TCTGTCCGTG CACACGTCTT
1201 TTCTCTCTCT CCTACGCAGT GGATTAATCG GCATGGCGGC TCTGGCCACG TCGCAGCTCG
1261 TCGCAACGCG CGCCGGCCTG GGCGTCCCGG ACGCGTCCAC GTTCCGCCGC GGCGCCGCGC
1321 AGGGCCTGAG GGGGGCCCGG GCGTCGGCGG CGGCGGACAC GCTCAGCATG CGGACCAGCG
1381 CGCGCGCGGC GCCCAGGCAC CAGCAGCAGG CGCGCCGCGG GGGCAGGTTC CCGTCGCTCG
1441 TCGTGTGCGC CAGCGCCGGC ATGAACGTCG TCTTCGTCGG CGCCGAGATG GCGCCGTGGA
1501 GCAAGACCGG CGGCCTCGGG GACGTCCTCG GCGGCCTGCC GCCGGCCATG GCCGTAAGCG
1561 CGCGCACCGA GACATGCATC CGTTGGATCG CGTCTTCTTC GTGCTCTTGC CGCGTGCATG
1621 ATGCATGTGT TTCCTCCTGG CTTGTGTTCG TGTATGTGAC GTGTTTGTTC GGGCATGCAT
1681 GCAGGCGAAC GGGCACCGTG TCATGGTCGT CTCTCCCCGC TACGACCAGT ACAAGGACGC
1741 CTGGGACACC AGCGTCGTGT CCGAGGTACG GCCACCGAGA CCAGATTCAG ATCACAGTCA
1801 CACACACCGT CATATGAACC TTTCTCTGCT CTGATGCCTG CAACTGCAAA TGCATGCAGA
1861 TCAAGATGGG AGACGGGTAC GAGACGGTCA GGTTCTTCCA CTGCTACAAG CGCGGAGTGG
1921 ACCGCGTGTT CGTTGACCAC CCACTGTTCC TGGAGAGGGT GAGACGAGAT CTGATCACTC
1981 GATACGCAAT TACCACCCCA TTGTAAGCAG TTACAGTGAG CTTTTTTTCC CCCCGGCCTG
2041 GTCGCTGGTT TCAGGTTTGG GGAAAGACCG AGGAGAAGAT CTACGGGCCT GTCGCTGGAA
2101 CGGACTACAG GGACAACCAG CTGCGGTTCA GCCTGCTATG CCAGGTCAGG ATGGCTTGGT
2161 ACTACAACTT CATATCATCT GTATGCAGCA GTATACACTG ATGAGAAATG CATGCTGTTC
2221 TGCAGGCAGC ACTTGAAGCT CCAAGGATCC TGAGCCTCAA CAACAACCCA TACTTCTCCG
2281 GACCATACGG TAAGAGTTGC AGTCTTCGTA TATATATCTG TTGAGCTCGA GAATCTTCAC
2341 AGGAAGCGGC CCATCAGACG GACTGTCATT TTACACTGAC TACTGCTGCT GCTCTTCGTC
2401 CATCCATACA AGGGGAGGAC GTCGTGTTCG TCTGCAACGA CTGGCACACC GGCCCTCTCT
2461 CGTGCTACCT CAAGAGCAAC TACCAGTCCC ACGGCATCTA CAGGGACGCA AAGGTTGCCT
2521 TCTCTGAACT GAACAACGCC GTTTTCGTTC TCCATGCTCG TATATACCTC GTCTGGTAGT
2581 GGTGGTGCTT CTCTGAGAAA CTAACTGAAA CTGACTGCAT GTCTGTCTGA CCATCTTCAC
2641 GTACTACCAG ACCGCTTTCT GCATCCACAA CATCTCCTAC CAGGGCCGGT TCGCCTTCTC
2701 CGACTACCCG GAGCTGAACC TCCCGGAGAG ATTCAAGTCG TCCTTCGATT TCATCGACGG
2761 GTCTGTTTTC CTGCGTGCAT GTGAACATTC ATGAATGGTA ACCCACAACT GTTCGCGTCC
2821 TGCTGGTTCA TTATCTGACC TGATTGCATT ATTGCAGCTA CGAGAAGCCC GTGGAAGGCC
2881 GGAAGATCAA CTGGATGAAG GCCGGGATCC TCGAGGCCGA CAGGGTCCTC ACCGTCAGCC
2941 CCTACTACGC CGAGGAGCTC ATCTCCGGCA TCGCCAGGGG CTGCGAGCTC GACAACATCA
3001 TGCGCCTCAC CGGCATCACC GGCATCGTCA ACGGCATGGA CGTCAGCGAG TGGGACCCCA
3061 GCAGGGACAA GTACATCGCC GTGAAGTACG ACGTGTCGAC GGTGAGCTGG CTAGCTCTGA
3121 TTCTGCTGCC TGGTCCTCCT GCTCATCATG CTGGTTCGGT ACTGACGCGG CAAGTGTACG
3181 TACGTGCGTG CGACGGTGGT GTCCGGTTCA GGCCGTGGAG GCCAAGGCGC TGAACAAGGA
3241 GGCGCTGCAG GCGGAGGTCG GGCTCCCGGT GGACCGGAAC ATCCCGCTGG TGGCGTTCAT
3301 CGGCAGGCTG GAAGAGCAGA AGGGCCCCGA CGTCATGGCG GCCGCCATCC CGCAGCTCAT
3361 GGAGATGGTG GAGGACGTGC AGATCGTTCT GCTGGTACGT GTGCGCCGGC CGCCACCCGG
3421 CTACTACATG CGTGTATCGT TCGTTCTACT GGAACATGCG TGTGAGCAAC GCGATGGATA
3481 ATGCTGCAGG GCACGGGCAA GAAGAAGTTC GAGCGCATGC TCATGAGCGC CGAGGAGAAG
3541 TTCCCAGGCA AGGTGCGCGC CGTGGTCAAG TTCAACGCGG CGCTGGCGCA CCACATCATG
3601 GCCGGCGCCG ACGTGCTCGC CGTCACCAGC CGCTTCGAGC CCTGCGGCCT CATCCAGCTG
3661 CAGGGGATGC GATACGGAAC GGTACGAGAG AAAAAAAAAA TCCTGAATCC TGACGAGAGG
3721 GACAGAGACA GATTATGAAT GCTTCATCGA TTTGAATTGA TTGATCGATG TCTCCCGCTG
3781 CGACTCTTGC AGCCCTGCGC CTGCGCGTCC ACCGGTGGAC TCGTCGACAC CATCATCGAA
3841 GGCAAGACCG GGTTCCACAT GGGCCGCCTC AGCGTCGACG TAAGCCTAGC TCTGCCATGT
3901 TCTTTCTTCT TTCTTTCTGT ATGTATGTAT GAATCAGCAC CGCCGTTCTT GTTTCGTCGT
3961 CGTCCTCTCT TCCCAGTGTA ACGTCGTGGA GCCGGCGGAC GTCAAGAAGG TGGCCACCAC
4021 ATTGCAGCGC GCCATCAAGG TGGTCGGCAC GCCGGCGTAC GAGGAGATGG TGAGGAACTG
4081 CATGATCCAG GATCTCTCCT GGAAGGTACG TACGCCCGCC CCGCCCCGCC CCGCCAGAGC
4141 AGAGCGCCAA GATCGACCGA TCGACCGACC ACACGTACGC GCCTCGCTCC TGTCGCTGAC
4201 CGTGGTTTAA TTTGCGAAAT GCGCAGGGCC CTGCCAAGAA CTGGGAGAAC GTGCTGCTCA
4261 GCCTCGGGGT CGCCGGCGGC GAGCCAGGGG TCGAAGGCGA GGAGATCGCG CCGCTCGCCA
4321 AGGAGAACGT GGCCGCGCCC TGAAGAGTTC GGCCTGCAGG GCCCCTGATC TCGCGCGTGG
4381 TGCAAAGATG TTGGGACATC TTCTTATATA TGCTGTTTCG TTTATGTGAT ATGGACAAGT
4441 ATGTGTAGCT GCTTGCTTGT GCTAGTGTAA TGTAGTGTAG TGGTGGCCAG TGGCACAACC
4501 TAATAAGCGC ATGAACTAAT TGCTTGCGTG TGTAGTTAAG TACCGATCGG TAATTTTATA
4561 TTGCGAGTAA ATAAATGGAC CTGTAGTGGT GGAGTAAATA ATCCCTGCTG TTCGGTGTTC
4621 TTATCGCTCC TCGTATAGAT ATTATATAGA GTACATTTTT CTCTCTCTGA ATCCTACGTT
4681 TGTGAAATTT CTATATCATT ACTGTAAAAT TTCTGCGTTC CAAAAGAGAC CATAGCCTAT
4741 CTTTGGCCCT GTTTGTTTCG GCTTCTGGCA GCTTCTGGCC ACCAAAAGCT GCTGCGGACT
表1b
稻waxv基因的DNA序列和推定的氨基酸序列
[序列6和序列7]基因座 QSWX 2542bp RNA PLN定义 O.aativa Waxy mRNA.编号 X62134 S39554关键词 葡糖基转移酶;淀粉生物合成;蜡质基因来源 稻生物体 Eukaryota;Plantae; Embryobionta;Magnoliophyta;Liliopsida;
Commelinidae;Cyperales;Poaceae.参考文献 1 (bases 1 to 2542)
作者 Okayaki,R.J.
题目 Direct Submission
杂志 Submitted(12-SEP-1991)to the EMBL/GanBank/DDBJ databaees.R.J.
Okayaki,University of Florida,Dep of Vegetable Crops,1255
Fifield Hall,514 IFAS,Gainesville,Florida 32611-0514,USASTANDARD full automatic参考文献 2 (bases 1 to 2542)
作者 okagaki,R.J.
题目 Nucleotide sequence of a long cDNA from the rice waxy gene
杂志 Plant Mol.Biol.19,513-516(1992)
标准 full automatic注释 NCBI gi:20402特征 Locatlon/Qualifiers
来源 1..2542
/organism="Oryza sativa"
/dev stage="immature seed"
/tissue type="seed"
CDS 453..2282
/gene="Wx"
/standard_name="Waxy gene"
/EC_number="2.4.1.21"
/note="NCBI gi:20403"
/codon start=1
/functon="starch biosynthesis"
/product="starch(bacterial glycogen) synthase"/translation="MSALTTSQLATSATGFGIADRSAPSSLLRHGFQGLKPRSPAGGDATSLSVTTSARATPKQQRSVQRGSRRFPSVVVYATGAGMNVVFVGAEMAPWSKTGGLGDVLGGLPPAMAANGHRVMVISPRYDQYKDAWDTSVVAEIKVADRYERVRFFHCYKRGVDRVFIDHPSFLEKVWGKTGEKIYGPDTGVDYKDNQMRFSLLCQAALEAPRILNLNNNPYFKGTYGFDVVFVCNDWHTGPLASYLKNNYQPNGIYRNAKVAFCIHNISYQGRFAFEDYPELNLSERFRSSFDFIDGYDTPVEGRKINWMKAGILEADRVLTVSPYYAEELISGIARGCELDNIMRLTGITGIVNGMDVSEWDPSKDKYITAKYDATTAIEAKALNKEALQAEAGLPVDRKIPLIAFIGRLEEQKGPDVMAAAIPELMQEDVQIVLLGTGKKKFEKLLKSMEEKYPGKVRAVVKFNAPLAHLIMAGADVLAVPSRFEPCGLIQLQGMRYGTPCACASTGGLVDTVIEGKTGFHMGRLSVDCKVVEPSDVKKVAATLKRAIKVVGTPAYEEMVRNCMNQ
DLSWKGPAKNWENVLLGLGVAGSAPGIEGDEIAPLAKENVAAP"
3’UTR 2283..2535
polyA_site 2535BASE COUNT 610A 665C 693G 574TORIGIN
1 GAATTCAGTG TGAAGGAATA GATTCTCTTC AAAACAATTT AATCATTCAT CTGATCTGCT
61 CAAAGCTCTG TGCATCTCCG GGTGCAACGG CCAGGATATT TATTGTGCAG TAAAAAAATG
121 TCATATCCCC TAGCCACCCA AGAAACTGCT CCTTAAGTCC TTATAAGCAC ATATGGCATT
181 GTAATATATA TGTTTGAGTT TTAGCGACAA TTTTTTTAAA AACTTTTGGT CCTTTTTATG
241 AACGTTTTAA GTTTCACTGT CTTTTTTTTT CGAATTTTAA ATGTAGCTTC AAATTCTAAT
301 CCCCAATCCA AATTGTAATA AACTTCAATT CTCCTAATTA ACATCTTAAT TCATTTATTT
361 GAAAACCAGT TCAAATTCTT TTTAGGCTCA CCAAACCTTA AACAATTCAA TTCAGTGCAG
421 AGATCTTCCA CAGCAACAGC TAGACAACCA CCATGTCGGC TCTCACCACG TCCCAGCTCG
481 CCACCTCGGC CACCGGCTTC GGCATCGCCG ACAGGTCGGC GCCGTCGTCG CTGCTCCGCC
541 ACGGGTTCCA GGGCCTCAAG CCCCGCAGCC CCGCCGGCGG CGACGCGACG TCGCTCAGCG
601 TGACGACCAG CGCGCGCGCG ACGCCCAAGC AGCAGCGGTC GGTGCAGCGT GGCAGCCGGA
661 GGTTCCCCTC CGTCGTCGTG TACGCCACCG GCGCCGGCAT GAACGTCGTG TTCGTCGGCG
721 CCGAGATGGC CCCCTGGAGC AAGACCGGCG GCCTCGGTGA CGTCCTCGGT GGCCTCCCCC
781 CTGCCATGGC TGCGAATGGC CACAGGGTCA TGGTGATCTC TCCTCGGTAC GACCAGTACA
841 AGGACGCTTG GGATACCAGC GTTGTGGCTG AGATCAAGGT TGCAGACAGG TACGAGAGGG
901 TGAGGTTTTT CCATTGCTAC AAGCGTGGAG TCGACCGTGT GTTCATCGAC CATCCGTCAT
961 TCCTGGAGAA GGTTTGGGGA AAGACCGGTG AGAAGATCTA CGGACCTGAC ACTGGAGTTG
1021 ATTACAAAGA CAACCAGATG CGTTTCAGCC TTCTTTGCCA GGCAGCACTC GAGGCTCCTA
1081 GGATCCTAAA CCTCAACAAC AACCCATACT TCAAAGGAAC TTATGGTGAG GATGTTGTGT
1141 TCGTCTGCAA CGACTGGCAC ACTGGCCCAC TGGCGAGCTA CCTGAAGAAC AACTACCAGC
1201 CCAATGGCAT CTACAGGAAT GCAAAGGTTG CTTTCTGCAT CCACAACATC TCCTACCAGG
1261 GCCGTTTCGC TTTCGAGGAT TACCCTGAGC TGAACCTCTC CGAGAGGTTC AGGTCATCCT
1321 TCGATTTCAT CGACGGGTAT GACACGCCGG TGGAGGGCAG GAAGATCAAC TGGATGAAGG
1381 CCGGAATCCT GGAAGCCGAC AGGGTGCTCA CCGTGAGCCC GTACTACGCC GAGGAGCTCA
1441 TCTCCGGCAT CGCCAGGGGA TGCGAGCTCG ACAACATCAT GCGGCTCACC GGCATCACCG
1501 GCATCGTCAA CGGCATGGAC GTCAGCGAGT GGGATCCTAG CAAGGACAAG TACATCACCG
1561 CCAAGTACGA CGCAACCACG GCAATCGAGG CGAAGGCGCT GAACAAGGAG GCGTTGCAGG
1621 CGGAGGCGGG TCTTCCGGTC GACAGGAAAA TCCCACTGAT CGCGTTCATC GGCAGGCTGG
1681 AGGAACAGAA GGGCCCTGAC GTCATGGCCG CCGCCATCCC GGAGCTCATG CAGGAGGACG
1741 TCCAGATCGT TCTTCTGGGT ACTGGAAAGA AGAAGTTCGA GAAGCTGCTC AAGAGCATGG
1801 AGGAGAAGTA TCCGGGCAAG GTGAGGGCGG TGGTGAAGTT CAACGCGCCG CTTGCTCATC
1861 TCATCATGGC CGGAGCCGAC GTGCTCGCCG TCCCCAGCCG CTTCGAGCCC TGTGGACTCA
1921 TCCAGCTGCA GGGGATGAGA TACGGAACGC CCTGTGCTTG CGCGTCCACC GGTGGGCTCG
1981 TGGACACGGT CATCGAAGGC AAGACTGGTT TCCACATGGG CCGTCTCAGC GTCGACTGCA
2041 AGGTGGTGGA GCCAAGCGAC GTGAAGAAGG TGGCGGCCAC CCTGAAGCGC GCCATCAAGG2101 TCGTCGGCAC GCCGGCGTAC GAGGAGATGG TCAGGAACTG CATGAACCAG GACCTCTCCT2161 GGAAGGGGCC TGCGAAGAAC TGGGAGAATG TGCTCCTGGG CCTGGGCGTC GCCGGCAGCG2221 CGCCGGGGAT CGAAGGCGAC GAGATCGCGC CGCTCGCCAA GGAGAACGTG GCTGCTCCTT2281 GAAGAGCCTG AGATCTACAT ATGGAGTGAT TAATTAATAT AGCAGTATAT GGATGAGAGA2341 CGAATGAACC AGTGGTTTGT TTGTTGTAGT GAATTTGTAG CTATAGCCAA TTATATAGGC2401 TAATAAGTTT GATGTTGTAC TCTTCTGGGT GTGCTTAAGT ATCTTATCGG ACCCTGAATT2461 TATGTGTGTG GCTTATTGCC AATAATATTA AGTAATAAAG GGTTTATTAT ATTATTATAT2521 ATGTTATATT ATACTAAAAA AA
表2玉米可溶性淀粉合成酶IIa基因的DNA序列和推定的氨基酸序列
[序列8和序列9]文件名:MSS2C.SEQ 序列:正常2007BP密码子表:UNIV.TCN序列范围:1-2007翻译范围:1-2007★★★DNA翻译★★★1 GCT GAG GCT GAG GCC GGG GGC AAG GAC GCG CCG CCG GAG AGG AGC GGC 481 A E A E A G G K D A P P E R S G 1649 GAC GCC GCC AGG TTG CCC CGC GCT CGG CGC AAT GCG GTC TCC AAA CGG 9617 D A A R L P R A R R N A V S K R 3297 AGG GAT CCT CTT CAG CCG GTC GGC CGG TAC GGC TCC GCG ACG GGA AAC 14433 R D P L Q P V G R Y G S A T G N 48145 ACG GCC AGG ACC GGC GCC GCG TCC TGC CAG AAC GCC GCA TTG GCG GAC 19249 T A R T G A A S C Q N A A L A D 64193 GTT GAG ATC GTT GAG ATC AAG TCC ATC GTC GCC GCG CCG CCG ACG AGC 24065 V E I V E I K S I V A A P P T S 80241 ATA GTG AAG TTC CCA GGG CGC GGG CTA CAG GAT GAT CCT TCC CTC TGG 28881 I V K F P G R G L Q D D P S L W 96289 GAC ATA GCA CCG GAG ACT GTC CTC CCA GCC CCG AAG CCA CTG CAT GAA 33697 D I A P E T V L P A P K P L H E 112337 TCG CCT GCG GTT GAC GGA GAT TCA AAT GGA ATT GCA CCT CCT ACA GTT 384113 S P A V D G D S N G I A P P T V 128385 GAG CCA TTA GTA CAG GAG GCC ACT TGG GAT TTC AAG AAA TAC ATC GGT 432129 E P L V Q E A T W D F K K Y I G 144433 TTT GAC GAG CCT GAC GAA GCG AAG GAT GAT TCC AGG GTT GGT GCA GAT 480 145 F D E P D E A K D D S R V G A D 160481 GAT GCT GGT TCT TTT GAA CAT TAT GGG ACA ATG ATT CTG GGC CTT TGT 528161 D A G S F E H Y G T M I L G L C 176529 GGG GAG AAT GTT ATG AAC GTG ATC GTG GTG GCT GCT GAA TGT TCT CCA 576177 G E N V M N V I V V A A E C S P 192577 TGG TGC AAA ACA GGT GGT CTT GGA GAT GTT GTG GGA GCT TTA CCC AAG 624193 W C K T G G L G D V V G A L P K 208625 GCT TTA GCG AGA AGA GGA CAT CGT GTT ATG GTT GTG GTA CCA AGG TAT 672209 A L A R R G H R V M V V V P R Y 224673 GGG GAC TAT GTG GAA GCC TTT GAT ATG GGA ATC CGG AAA TAC TAC AAA 720225 G D Y V E A F D M G I R K Y Y K 240721 GCT GCA GGA CAG GAC CTA GAA GTG AAC TAT TTC CAT GCA TTT ATT GAT 768241 A A G Q D L E V N Y F H A F I D 256769 GGA GTC GAC TTT GTG TTC ATT GAT GCC TCT TTC CGG CAC CGT CAA GAT 816257 G V D F V F I D A S F R H R Q D 272817 GAC ATA TAT GGG GGA AGT AGG CAG GAA ATC ATG AAG CGC ATG ATT TTG 864273 D I Y G G S R Q E I M K R M I L 288865 TTT TGC AAG GTT GCT GTT GAG GTT CCT TGG CAC GTT CCA TGC GGT GGT 912289 F C K V A V E V P W H V P C G G 304913 GTG TGC TAC GGA GAT GGA AAT TTG GTG TTC ATT GCC ATG AAT TGG CAC 960305 V C Y G D G N L V F I A M N W H 320961 ACT GCA CTC CTG CCT GTT TAT CTG AAG GCA TAT TAC AGA GAC CAT GGG 1008321 T A L L P V Y L K A Y Y R D H G 3361009 TTA ATG CAG TAC ACT CGC TCC GTC CTC GTC ATA CAT AAC ATC GGC CAC 1056337 L M Q Y T R S V L V I H N I G H 3521057 CAG GGC CGT GGT CCT GTA CAT GAA TTC CCG TAC ATG GAC TTG CTG AAC 1104353 Q G R G P V H E F P Y M D L L N 3681105 ACT AAC CTT CAA CAT TTC GAG CTG TAC GAT CCC GTC GGT GGC GAG CAC 1152369 T N L Q H F E L Y D P V G G E H 3841153 GCC AAC ATC TTT GCC GCG TGT GTT CTG AAG ATG GCA GAC CGG GTG GTG 1200385 A N I F A A C V L K M A D R V V 4001201 ACT GTC AGC CGC GGC TAC CTG TGG GAG CTG AAG ACA GTG GAA GGC GGC 1248401 T V S R G Y L W E L K T V E G G 4161249 TGG GGC CTC CAC GAC ATC ATC CGT TCT AAC GAC TGG AAG ATC AAT GGC 1296417 W G L H D I I R S N D W K I N G 4321297 ATT CGT GAA CGC ATC GAC CAC CAG GAG TGG AAC CCC AAG GTG GAC GTG 1344433 I R E R I D H Q E W N P K V D V 4481345 CAC CTG CGG TCG GAC GGC TAC ACC AAC TAC TCC CTC GAG ACA CTC GAC 1392449 H L R S D G Y T N Y S L E T L D 4641393 GCT GGA AAG CGG CAG TGC AAG GCG GCC CTG CAG CGG GAC GTG GGC CTG 1440465 A G K R Q C K A A L Q R D V G L 4801441 GAA GTG CGC GAC GAC GTG CCG CTG CTC GGC TTC ATC GGG CGT CTG GAT 1488481 E V R D D V P L L G F I G R L D 4961489 GGA CAG AAG GGC GTG GAC ATC ATC GGG GAC GCG ATG CCG TGG ATC GCG 1536497 G Q K G V D I I G D A M P W I A 5121537 GGG CAG GAC GTG CAG CTG GTG ATG CTG GGC ACC GGC CCA CCT GAC CTG 1584513 G Q D V Q L V M L G T G P P D L 5281585 GAA CGA ATG CTG CAG CAC TTG GAG CGG GAG CAT CCC AAC AAG GTG CGC 1632529 E R M L Q H L E R E H P N K V R 5441633 GGG TGG GTC GGG TTC TCG GTC CTA ATG GTG CAT CGC ATC ACG CCG GGC 1680545 G W V G F S V L M V H R I T P G 5601681 GCC AGC GTG CTG GTG ATG CCC TCC CGC TTC GCC GGC GGG CTG AAC CAG 1728561 A S V L V M P S R F A G G L N Q 5761729 CTC TAC GCG ATG GCA TAC GGC ACC GTC CCT GTG GTG CAC GCC GTG GGC 1776577 L Y A M A Y G T V P V V H A V G 5921777 GGG CTC AGG GAC ACC GTG GCG CCG TTC GAC CCG TTC GGC GAC GCC GGG 1824593 G L R D T V A P F D P F G D A G 6081825 CTC GGG TGG ACT TTT GAC CGC GCC GAG GCC AAC AAG CTG ATC GAG GTG 1872609 L G W T F D R A E A N K L I E V 6241873 CTC AGC CAC TGC CTC GAC ACG TAC CGA AAC TAC GAG GAG AGC TGG AAG 1920625 L S H C L D T Y R N Y E E S W K 6401921 AGT CTC CAG GCG CGC GGC ATG TCG CAG AAC CTC AGC TGG GAC CAC GCG 1968641 S L Q A R G M S Q N L S W D H A 6561969 GCT GAG CTC TAC GAG GAC GTC CTT GTC AAG TAC CAG TGG 2007657 A E L Y E D V L V K Y Q W 669
表3玉米可溶性淀粉合成酶IIb基因的DNA序列和推定的氨基酸序列
[序列10和序列11]文件名:MSS3FULL.DNA 序列:正常2097BP密码子表:UNIV.TCN序列范围:1-2097翻译范围:1-2097★★★DNA翻译★★★1 ATG CCG GGG GCA ATC TCT TCC TCG TCG TCG GCT TTT CTC CTC CCC GTC 481 M P G A I S S S S S A F L L P V 1649 GCG TCC TCC TCG CCG CGG CGC AGG CGG GGC AGT GTG GGT GCT GCT CTG 9617 A S S S P R R R R G S V G A A L 3297 CGC TCG TAC GGC TAC AGC GGC GCG GAG CTG CGG TTG CAT TGG GCG CGG 14433 R S Y G Y S G A E L R L H W A R 48145 CGG GGC CCG CCT CAG GAT GGA GCG GCG TCG GTA CGC GCC GCA GCG GCA 19249 R G P P Q D G A A S V R A A A A 64193 CCG GCC GGG GGC GAA AGC GAG GAG GCA GCG AAG AGC TCC TCC TCG TCC 24065 P A G G E S E E A A K S S S S S 80241 CAG GCG GGC GCT GTT CAG GGC AGC ACG GCC AAG GCT GTG GAT TCT GCT 288 81 Q A G A V Q G S T A K A V D S A 96289 TCA CCT CCC AAT CCT TTG ACA TCT GCT CCG AAG CAA AGT CAG AGC GCT 33697 S P P N P L T S A P K Q S Q S A 112337 GCA ATG CAA AAC GGA ACG AGT GGG GCC AGC AGC GCG AGC ACC GCC GCG 384113 A M Q N G T S G G S S A S T A A 128385 CCG GTG TCC GGA CCC AAA GCT GAT CAT CCA TCA GCT CCT GTC ACC AAG 432129 P V S G P K A D H P S A P V T K 144433 AGA GAA ATC GAT GCC AGT GCG GTG AAG CCA GAG CCC GCA GGT GAT GAT 480145 R E I D A S A V K P E P A G D D 160481 GCT AGA CCG GTG GAA AGC ATA GGC ATC GCT GAA CCG GTG GAT GCT AAG 528161 A R P V E S I G I A E P V D A K 176529 GCT GAT GCA GCT CCG GCT ACA GAT GCG GCG GCG AGT GCT CCT TAT GAC 576177 A D A A P A T D A A A S A P Y D 192577 AGG GAG GAT AAT GAA CCT GGC CCT TTG GCT GGG CCT AAT GTG ATG AAC 624193 R E D N E P G P L A G P N V M N 208625 GTC GTC GTG GTG GCT TCT GAA TGT GCT CCT TTC TGC AAG ACA GGT GGC 672209 V V V V A S E C A P F C K T G G 224673 CTT GGA GAT GTC GTG GGT GCT TTG CCT AAG GCT CTG GCG AGG AGA GGA 720225 L G D V V G A L P K A L A R R G 240721 CAC CGT GTT ATG GTC GTG ATA GCA AGA TAT GGA GAG TAT GCC GAA GCC 768241 H R V M V V I P R Y G E Y A E A 256769 CGG GAT TTA GGT GTA AGG AGA CGT TAC AAG GTA GCT GGA CAG GAT TCA 816257 R D L G V R R R Y K V A G Q D S 272817 GAA GTT ACT TAT TTT CAC TCT TAC ATT GAT GGA GTT GAT TTT GTA TTC 864273 E V T Y F H S Y I D G V D F V F 288865 GTA GAA GCC CCT CCC TTC CGG CAC CGG CAC AAT AAT ATT TAT GGG GGA 912289 V E A P P F R H R H N N I Y G G 304913 GAA AGA TTG GAT ATT TTG AAG CGC ATG ATT TTG TTC TGC AAG GCC GCT 960305 E R L D I L K R M I L F C K A A 320961 GTT GAG GTT CCA TGG TAT GCT CCA TGT GGC GGT ACT GTC TAT GGT GAT 1008321 V E V P W Y A P C G G T V Y G D 3361009 GGC AAC TTA GTT TTC ATT GCT AAT GAT TGG CAT ACC GCA CTT CTG CCT 1056337 G N L V F I A N D W H T A L L P 3521057 GTC TAT CTA AAG GCC TAT TAC CGG GAC AAT GGT TTG ATG CAG TAT GCT 1104353 V Y L K A Y Y R D N G L M Q Y A 3681105 CGC TCT GTG CTT GTG ATA CAC AAC ATT GCT CAT CAG GGT CGT GGC CCT 1152369 R S V L V I H N I A H Q G R G P 3841153 GTA GAC GAC TTC GTC AAT TTT GAC TTG CCT GAA CAC TAC ATC GAC CAC 1200385 V D D F V N F D L P E H Y I D H 4001201 TTC AAA CTG TAT GAC AAC ATT GGT GGG GAT CAC AGC AAC GTT TTT GCT 1248401 F K L Y D N I G G D H S N V F A 4161249 GCG GGG CTG AAG ACG GCA GAC CGG GTG GTG ACC GTT AGC AAT GGC TAC 1296417 A G L K T A D R V V T V S N G Y 4321297 ATG TGG GAG CTG AAG ACT TCG GAA GGC GGG TGG GGC CTC CAC GAC ATC 1344433 M W E L K T S E G G W G L H D I 4481345 ATA AAC CAG AAC GAC TGG AAG CTG CAG GGC ATC GTG AAC GGC ATC GAC 1392449 I N Q N D W K L Q G I V N G I D 4641393 ATG AGC GAG TGG AAC CCC GCT GTG GAC GTG CAC CTC CAC TCC GAC GAC 1440465 M S E W N P A V D V H L H S D D 4801441 TAC ACC AAC TAC ACG TTC GAG ACG CTG GAC ACC GGC AAG CGG CAG TGC 1488481 Y T N Y T F E T L D T G K R Q C 4961489 AAG GCC GCC CTG CAG CGG CAG CTG GGC CTG CAG GTC CGC GAC GAC GTG 1536497 K A A L Q R Q L G L Q V R D D V 5121537 CCA CTG ATC GGG TTC ATC GGG CGG CTG GAC CAC CAG AAG GGC GTG GAC 1584513 P L I G F I G R L D H Q K G V D 5281585 ATC ATC GCC GAC GCG ATC CAC TGG ATC GCG GGG CAG GAC GTG CAG CTC 632529 I I A D A I H W I A G Q D V Q L 5441633 GTG ATG CTG GGC ACC GGG CGG GCC GAC CTG GAG GAC ATG CTG CGG CGG 1680545 V M L G T G R A D L E D M L R R 5601681 TTC GAG TCG GAG CAC AGC GAC AAG GTG CGC GCG TGG GTG GGG TTC TCG 1728561 F E S E H S D K V R A W V G F S 5761729 GTG CCC CTG GCG CAC CGC ATC ACG GCG GGC GCG GAC ATC CTG CTG ATG 1776577 V P L A H R I T A G A D I L L M 5921777 CCG TCG CGG TTC GAG CCG TGC GGG CTG AAC CAG CTC TAC GCC ATG GCG 1824593 P S R F E P C G L N Q L Y A M A 6081825 TAC GGG ACC GTG CCC GTG GTG CAC GCC GTG GGG GGG CTC CGG GAC ACG 1872609 Y G T V P V V H A V G G L R D T 6241873 GTG GCG CCG TTC GAC CCG TTC AAC GAC ACC GGG CTC GGG TGG ACG TTC 1920625 V A P F D P F N D T G L G W T F 6401921 GAC CGC GCG GAG GCG AAC CGG ATG ATC GAC GCG CTC TCG CAC TGC CTC 1968641 D R A E A N R M I D A L S H C L 6561969 ACC ACG TAC CGG AAC TAC AAG GAG AGC TGG CGC GCC TGC AGG GCG CGC 2016657 T T Y R N Y K E S W R A C R A R 6722017 GGC ATG GCC GAG GAC CTC AGC TGG GAC CAC GCC GCC GTG CTG TAT GAG 2064673 G M A E D L S W D H A A V L Y E 6882065 GAC GTG CTC GTC AAG GCG AAG TAC CAG TGG TGA 2097689 D V L V K A K Y Q W ★ 699
表4玉米可溶性淀粉合成酶I基因的DNA序列和推定的氨基酸序列
[序列12和序列13]文件名:MSS3FULL.DNA 序列:正常1752BP密码子表:UNIV.TCN序列范围:1-1752翻译范围:1-1752★★★DNA翻译★★★TGC GTC GCG GAG CTG AGC AGG GAG GGG CCC GCG CCG CGC CCG CTG CCA 48Cys Val Ala Glu Leu Ser Arg Glu Gly Pro Ala Pro Arg Pro Leu Pro700 705 710 715CCC GCG CTG CTG GCG CCC CCG CTC GTG CCC GGC TTC CTC GCG CCG CCG 96Pro Ala Leu Leu Ala Pro Pro Leu Val Pro Gly Phe Leu Ala Pro Pro
720 725 730GCC GAG CCC ACG GGT GAG CCG GCA TCG ACG CCG CCG CCC GTG CCC GAC 144Ala Glu Pro Thr Gly Glu Pro Ala Ser Thr Pro Pro Pro Val Pro Asp
735 740 745GCC GGC CTG GGG GAC CTC GGT CTC GAA CCT GAA GGG ATT GCT GAA GGT 192Ala Gly Leu Gly Asp Leu GIy Leu Glu Pro Glu Gly Ile Ala Glu Gly
750 755 760TCC ATC GAT AAC ACA GTA GTT GTG GCA AGT GAG CAA GAT TCT GAG ATT 240Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln Asp Ser Glu Ile
765 770 775GTG GTT GGA AAG GAG CAA GCT CGA GCT AAA GTA ACA CAA AGC ATT GTC 288Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr Gln Ser Ile Val780 785 790 795TTT GTA ACC GGC GAA GCT TCT CCT TAT GCA AAG TCT GGG GGT CTA GGA 336Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser Gly Gly Leu Gly
800 805 810GAT GTT TGT GGT TCA TTG CCA GTT GCT CTT GCT GCT CGT GGT CAC CGT 384Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala Arg Gly His Arg
815 820 825GTG ATG GTT GTA ATG CCC AGA TAT TTA AAT GGT ACC TCC GAT AAG AAT 432Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr Ser Asp Lys Asn
830 835 840TAT GCA AAT GCA TTT TAC ACA GAA AAACAC ATT CGG ATT CCA TGC TTT 480Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg Iie Pro Cys Phe
845 850 855GGC GGT GAA CAT GAA GTT ACC TTC TTC CAT GAG TAT AGA GAT TCA GTT 528Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr Arg Asp Ser Val860 865 870 875GAC TGG GTG TTT GTT GAT CAT CCC TCA TAT CAC AGA CCT GGA AAT TTA 576Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg Pro Gly Asn Leu
880 885 890TAT GGA GAT AAG TTT GGT GCT TTT GGT GAT AAT CAG TTC AGA TAC ACA 624Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln Phe Arg Tyr Thr
895 900 905CTC CTT TGC TAT GCT GCA TGT GAG GCT CCT TTG ATC CTT GAA TTG GGA 672Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile Leu Glu Leu Gly
910 915 920GGA TAT ATT TAT GGA CAG AAT TGC ATG TTT GTT GTC AAT GAT TGG CAT 720Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val Asn Asp Trp His
925 930 935GCC AGT CTA GTG CCA GTC CTT CTT GCT GCA AAA TAT AGA CCA TAT GGT 768Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr Arg Pro Tyr Gly940 945 950 955GTT TAT AAA GAC TCC CGC AGC ATT CTT GTA ATA CAT AAT TTA GCA CAT 816Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile His Asn Leu Ala His
960 965 970CAG GGT GTA GAG CCT GCA AGC ACA TAT CCT GAC CTT GGG TTG CCA CCT 864Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu Gly Leu Pro Pro
975 980 985GAA TGG TAT GGA GCT CTG GAG TGG GTA TTC CCT GAA TGG GCG AGG AGG 912Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu Trp Ala Arg Arg
990 995 1000CAT GCC CTT GAC AAG GGT GAG GCA GTT AAT TTT TTG AAA GGT GCA GTT 960His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu Lys Gly Ala Val
1005 1010 1015GTG ACA GCA GAT CGA ATC GTG ACT GTC AGT AAG GGT TAT TCG TGG GAG 1008Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly Tyr Ser Trp Glu1020 1025 1030 1035GTC ACA ACT GCT GAA GGT GGA CAG GGC CTC AAT GAG CTC TTA AGC TCC 1056Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu Leu Leu Ser Ser
1040 1045 1050AGA AAG AGT GTA TTA AAC GGA ATT GTA AAT GGA ATT GAC ATT AAT GAT 1104Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile Asp Ile Asn Asp
1055 1060 1065TGG AAC CCT GCC ACA GAC AAA TGT ATC CCC TGT CAT TAT TCT GTT GAT 1152Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His Tyr Ser Val Asp
1070 1075 1080GAC CTC TCT GGA AAG GCC AAA TGT AAA GGT GCA TTG CAG AAG GAG CTG 1200Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu Gln Lys Glu Leu
1085 1090 1095GGT TTA CCT ATA AGG CCT GAT GTT CCT CTG ATT GGC TTT ATT GGA AGG 1248Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly Phe Ile Gly Arg1100 1105 1110 1115TTG GAT TAT CAG AAA GGC ATT GAT CTC ATT CAA CTT ATC ATA CCA GAT 1296Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu Ile Ile Pro Asp
1120 1125 1130CTC ATG CGG GAA GAT GTT CAA TTT GTC ATG CTT GGA TCT GGT GAC CCA 1344Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly Ser Gly Asp Pro
1135 1140 1145GAG CTT GAA GAT TGG ATG AGA TCT ACA GAG TCG ATC TTC AAG GAT AAA 1392Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile Phe Lys Asp Lys
1150 1155 1160TTT CGT GGA TGG GTT GGA TTT AGT GTT CCA GTT TCC CAC CGA ATA ACT 1440Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser His Arg Ile Thr
1165 1170 1175GCC GGC TGC GAT ATA TTG TTA ATG CCA TCC AGA TTC GAA CCT TGT GGT 1488Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe Glu Pro Cys Gly1180 1185 1190 1195CTC AAT CAG CTA TAT GCT ATG CAG TAT GGC ACA GTT CCT GTT GTC CAT 1536Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val Pro Val Val His
1200 1205 1210GCA ACT GGG GGC CTT AGA GAT ACC GTG GAG AAC TTC AAC CCT TTC GGT 1584Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe Asn Pro Phe Gly
1215 1220 1225GAG AAT GGA GAG CAG GGT ACA GGG TGG GCA TTC GCA CCC CTA ACC ACA 1632Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala Pro Leu Thr Thr
1230 1235 1240GAA AAC ATG TTT GTG GAC ATT GCG AAC TGC AAT ATC TAC ATA CAG GGA 1680Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile Tyr Ile Gln Gly
1245 1250 1255ACA CAA GTC CTC CTG GGA AGG GCT AAT GAA GCG AGG CAT GTC AAA AGA 1728Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg His Val Lys Arg1260 1265 1270 1275CTT CAC GTG GGA CCA TGC CGC TGA 1752Leu His Val Gly Pro Cys Arg ★
1280(2)序列13资料:
(i)序列特征
(A)长度:584个氨基酸
(B)类型:氨基酸
(D)拓扑结构:线性
(ii)分子类型:蛋白
(xi)序列描述:序列13Cys Val Ala Glu Leu Ser Arg Glu Gly Pro Ala Pro Arg Pro Leu Pro1 5 10 15Pro Ala Leu Leu Ala Pro Pro Leu Val Pro Gly Phe Leu Ala Pro Pro
20 25 30Ala Glu Pro Thr Gly Glu Pro Ala Ser Thr Pro Pro Pro Val Pro Asp
35 40 45Ala Gly Leu Gly Asp Leu Gly Leu Glu Pro Glu Gly Ile Ala Glu Gly
50 55 60Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln Asp Ser Glu Ile65 70 75 80Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr Gln Ser Ile Val
85 90 95Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser Gly Gly Leu Gly
100 105 110Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala Arg Gly His Arg
115 120 125Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr Ser Asp Lys Asn
130 135 140Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg Ile Pro Cys Phe145 150 155 160Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr Arg Asp Ser Val
165 170 175Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg Pro Gly Asn Leu
180 185 190Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln Phe Arg Tyr Thr
195 200 205Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile Leu Glu Leu Gly
210 215 220Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val Asn Asp Trp His225 230 235 240Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr Arg Pro Tyr Gly
245 250 255Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile His Asn Leu Ala His
260 265 270Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu Gly Leu Pro Pro
275 280 285Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu Trp Ala Arg Arg
290 295 300His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu Lys Gly Ala Val305 310 315 320Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly Tyr Ser Trp Glu
325 330 335Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu Leu Leu Ser Ser
340 345 350Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile Asp Ile Asn Asp
355 360 365Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His Tyr Ser Val Asp
370 375 380Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu Gln Lys Glu Leu385 390 395 400Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly Phe Ile Gly Arg
405 410 415Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu Ile Ile Pro Asp
420 425 430Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly Ser Gly Asp Pro
435 440 445Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile Phe Lys Asp Lys
450 455 460Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser His Arg Ile Thr465 470 475 480Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe Glu Pro Cys Gly
485 490 495Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val Pro Val Val His
500 505 510Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe Asn Pro Phe Gly
515 520 525Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala Pro Leu Thr Thr
530 535 540Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile Tyr Ile Gln Gly545 550 555 560Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg His Val Lys Arg
565 570 575Leu His Val Gly Pro Cys Arg ★
580
表5玉米分支酶II基因和转运肽的mRNA序列和推定的氨基酸序列
[序列14和序列15]LOCUS MZEGLUCTRN 2725bp ss-mRNA PLN定义 Corn starch branching enzyme II mRNA,complete cds.编号 L08065关键词 1,4-alpha-glucan branching enzyme;amylo-transglycosylase;
glucanotransferase;starch branching enzyme II.来源 Zea mays cDNA to mRNA.生物体 Zea mays
Eukaryota;Plantae;Embryobionta;Magnoliophyta;Liliopsida;
Commelinidae;Cyperales;Poaceae.参考文献 1 (bases 1 to 2725)
作者 Fisher,D.K.,Boyer,C.D.and Hannah,L.C.
题目 Starch branching enzyme II from maize endosperm
杂志 Plant physiol.102,1045-1046(1993)
标准 full automatic注释 NCBI gi:168482特征 Location/Qualifiers
来源 1..2725
/Cultivar="W64A×182E"
/dev_stage="29 days post pollenation"
/tissue_type="endosperm"
/organism="zea mays"
sig_peptide 91..264
/Codon start=1
CDS 91..2490
/EC_number="2.4.1.18"
/note="NC8I gi:168483"
/codon start=1
/product="starch branching enzyme II"/translation="MAFRVSGAVLGGAVRAPRLTGGGEGSLVFRHTGLFLTRGARVGCSGTHGAMRAAAAARKAVMVPEGENDGLASRADSAQFQSDELEVPDISEETTCGAGVADAQALNRVRVVPPPSDGQKIFQIDPMLQGYKYHLEYRYSLYRRIRSDIDEHEGGLEAFSRSYEKFGFNASAEGITYREWAPGAFSAALVGDVNNWDPNADRMSKNEFGVWEIFLPNNADGTSPIPHGSRVKVRMDTPSGIKDSIPAWIKYSVQAPGEIPYDGIYYDPPEEVKYVFRHAQPKRPKSLRIYETHVGMSSPEPKINTYVNFRDEVLPRIKKLGYNAVQIMAIQEHSYYGSFGYHVTNFFAPSSRFGTPEDLKSLIDRAHELGLLVLMDVVHSHASSNTLDGLNGFDGTDTHYFHSGPRGHHWMWDSRLFNYGNWEVLRFLLSNARWWLEEYKFDGFRFDGVTSMMYTHHGLQVTFTGNFNEYFGFATDVDAVVYLMLVNDLIHGLYPEAVTIGEDVSGMPTFALPVHDGGVGFDYRMHMAVADKWIDLLKQSDETWKMGDIVHTLTNRRWLEKCVTYAESHDQALVGDKTIAFWLMDKDMYDFMALDRPSTPTIDRGIALHKMIRLITMGLGGEGYLNFMGNEFGHPEWIDFPRGPQRLPSGKFIPGNNNSYDKCRRRFDLGDADYLRYHGMQEFDQAMQHLEQKYEFMTSDHQYISRKHEEDKVIVFEKGDLVFVFNFHCNNSYFDYRIGCRKPGVYKVVLDSDAGLFGGFSRIHHAAEHFTADCSHDNRPYSFSVYTPSRTCVVYAPV
E"
mat_peptide 265..2487
/codon_start=1
/product="starch branching enzyme II"BA5E COUNT 727A 534 C 715 G 749 TORIGIN
1 GGCCCAGAGC AGACCCGGAT TTCGCTCTTG CGGTCGCTGG GGTTTTAGCA TTGGCTGATC
61 AGTTCGATCC GATCCGGCTG CGAAGGCGAG ATGGCGTTCC GGGTTTCTGG GGCGGTGCTC
121 GGTGGGGCCG TAAGGGCTCC CCGACTCACC GGCGGCGGGG AGGGTAGTCT AGTCTTCCGG
181 CACACCGGCC TCTTCTTAAC TCGGGGTGCT CGAGTTGGAT GTTCGGGGAC GCACGGGGCC
241 ATGCGCGCGG CGGCCGCGGC CAGGAAGGCG GTCATCGTTC CTGAGGGCGA GAATGATGGC
301 CTCGCATCAA GGGCTGACTC GGCTCAATTC CAGTCGGATG AACTGGAGGT ACCAGACATT
361 TCTGAAGAGA CAACGTGCGG TGCTGGTGTG GCTGATGCTC AAGCCTTGAA CAGAGTTCGA
421 GTGGTCCCCC CACCAAGCGA TGGACAAAAA ATATTCCAGA TTGACCCCAT GTTGCAAGGC
481 TATAAGTACC ATCTTGAGTA TCGGTACAGC CTCTATAGAA GAATCCGTTC AGACATTGAT
541 GAACATGAAG GAGGCTTGGA AGCCTTCTCC CGTAGTTATG AGAAGTTTGG ATTTAATGCC
601 AGCGCGGAAG GTATCACATA TCGAGAATGG GCTCCTGGAG CATTTTCTGC AGCATTGGTG
661 GGTGACGTCA ACAACTGGGA TCCAAATGCA GATCGTATGA GCAAAAATGA GTTTGGTGTT
721 TGGGAAATTT TTCTGCCTAA CAATTCAGAT GGTACATCAC CTATTCCTCA TGGATCTCGT
781 GTAAAGGTGA GAATGGATAC TCCATCAGGG ATAAAGGATT CAATTCCAGC CTGGATCAAG
841 TACTCAGTGC AGGCCCCAGG AGAAATACCA TATGATGGGA TTTATTATGA TCCTCCTGAA
901 GAGGTAAAGT ATGTGTTCAG GCATGCGCAA CCTAAACGAC CAAAATCATT GCGGATATAT
961 GAAACACATG TCGGAATGAG TAGCCCGGAA CCGAAGATAA ACACATATGT AAACTTTAGG
1021 GATGAAGTCC TCCCAAGAAT AAAAAAACTT GGATACAATG CAGTGCAAAT AATGGCAATC
1081 CAAGAGCACT CATATTATGG AAGCTTTGGA TACCATGTAA CTAATTTTTT TGCGCCAAGT
1141 AGTCGTTTTG GTACCCCAGA AGATTTGAAG TCTTTGATTG ATAGAGCACA TGAGCTTGGT
1201 TTGCTAGTTC TCATGGATGT GGTTCATAGT CATGCGTCAA GTAATACTCT GGATGGGTTG
1261 AATGGTTTTG ATGGTACAGA TACACATTAC TTTCACAGTG GTCCACGTGG CCATCACTGG
1321 ATGTGGGATT CTCGCCTATT TAACTATGGG AACTGGGAAG TTTTAAGATT TCTTCTCTCC
1381 AATGCTAGAT GGTGGCTCGA GGAATATAAG TTTGATGGTT TCCGTTTTGA TGGTGTGACC
1441 TCCATGATGT ACACTCACCA CGGATTACAA GTAACATTTA CGGGGAACTT CAATGAGTAT
1501 TTTGGCTTTG CCACCGATGT AGATGCAGTG GTTTACTTGA TGCTGGTAAA TGATCTAATT
1561 CATGGACTTT ATCCTGAGGC TGTAACCATT GGTGAAGATG TTAGTGGAAT GCCTACATTT
1621 GCCCTTCCTG TTCACGATGG TGGGGTAGGT TTTGACTATC GGATGCATAT GGCTGTGGCT
1681 GACAAATGGA TTGACCTTCT CAAGCAAAGT GATGAAACTT GGAAGATGGG TGATATTGTG
1741 CACACACTGA CAAATAGGAG GTGGTTAGAG AAGTGTGTAA CTTATGCTGA AAGTCATGAT
1801 CAAGCATTAG TCGGCGACAA GACTATTGCG TTTTGGTTGA TGGACAAGGA TATGTATGAT
1861 TTCATGGCCC TCGATAGACC TTCAACTCCT ACCATTGATC GTGGGATAGC ATTACATAAG
1921 ATGATTAGAC TTATCACAAT GGGTTTAGGA GGAGAGGGCT ATCTTAATTT CATGGGAAAT
1981 GAGTTTGGAC ATCCTGAATG GATAGATTTT CCAAGAGGTC CGCAAAGACT TCCAAGTGGT
2041 AAGTTTATTC CAGGGAATAA CAACAGTTAT GACAAATGTC GTCGAAGATT TGACCTGGGT
2101 GATGCAGACT ATCTTAGGTA TCATGGTATG CAAGAGTTTG ATCAGGCAAT GCAACATCTT
2161 GAGCAAAAAT ATGAATTCAT GACATCTGAT CACCAGTATA TTTCCCGGAA ACATGAGGAG
2221 GATAAGGTGA TTGTGTTCGA AAAGGGAGAT TTGGTATTTG TGTTCAACTT CCACTGCAAC
2281 AACAGCTATT TTGACTACCG TATTGGTTGT CGAAAGCCTG GGGTGTATAA GGTGGTCTTG
2341 GACTCCGACG CTGGACTATT TGGTGGATTT AGCAGGATCC ATCACGCAGC CGAGCACTTC
2401 ACCGCCGACT GTTCGCATGA TAATAGGCCA TATTCATTCT CGGTTTATAC ACCAAGCAGA
2461 ACATGTGTCG TCTATGCTCC AGTGGAGTGA TAGCGGGGTA CTCGTTGCTG CGCGGCATGT
2521 GTGGGGCTGT CGATGTGAGG AAAAACCTTC TTCCAAAACC GGCAGATGCA TGCATGCATG
2581 CTACAATAAG GTTCTGATAC TTTAATCGAT GCTGGAAAGC CCATGCATCT CGCTGCGTTG
2641 TCCTCTCTAT ATATATAAGA CCTTCAAGGT GTCAATTAAA CATAGAGTTT TCGTTTTTCG
2701 CTTTCCTAAA AAAAAAAAAA AAAAA
表6玉米分支酶I和转运肽的mRNA序列和推定的氨基酸序列
[序列16和序列17]
LOCUS MZEBEI 2763 bp ss-mRNA PLN
DEFINITION Mai2e mRNA for branching enzyme-I (BE-I).
ACCESSION D11081
KEYWORDS branching enzyme-I.
SOURCE Zea mays L.(inbred oh43),cDNA to mRNA.
ORGANISM Zea mays
Eukaryota;Plantae;Embryobionta;Magnoliophyta;Liliopsida;
Commelinidae;Liliopsida.
参考文献 1 (bases 1 to 2763)
作者 Baba,T.,Kimura,K.,Mizuno,K.,Etoh,H.,Ishida,Y.,Shida,O.and
Arai,Y.
题目 Sequence conservation of the catalytic regions of Amylolytic
enzymes in maize branching enzyme-I
杂志 Biochem.Biophys.Res.Commun.181,87-94(1991)
标准 full automatic注释 Submitted (30-APR-1992) to DDBJ by:Tadashi Baba
Institute of Applied Biochemistry
University of Tsukuba
Tsukuba,Ibaraki 305
Japan
Phone: 0298-53-6632
Fax: 0298-53-6632.
NCBI gi: 211959特征 Location/Qualifiers
来源 1..2763
/organism="Zea mays"
CDS <1..2470
/note="NCBI gi:217960"
/codon_start=2
/product="branching enzyme-I precursor"/translation="LCLVSPSSSPTPLPPPRRSRSHADRAAPPGIAGGGNVRLSVLSVQCKARRSGVRKVKSKFATAATVQEDKTMATAKGDVDHLPIYDLDPKLEIFKDHFRYRMKRFLEQKGSIEENEGSLESFSKGYLKFGINTNEDGTVYREWAPAAQEAELIGDFNDWNGANHKMEKDKFGVWSIKIDHVKGKPAIPHNSKVKFRFLHGGVWVDRIPALIRYATVDASKFGAPYDGVHWDPPASERYTFKHPRPSKPAAPRIYEAHVGMSGEKPAVSTYREFADNVLPRIRANNYNTVQLMAVMEHSYYASFGYHVTNFFAVSSRSGTPEDLKYLVDKAHSLGLRVLMDVVHSHASNNVTDGLNGYDVGQSTQESYFHAGDRGYHKLWDSRLFNYANWEVLRFLLSNLRYWLDEFMFDGFRFDGVTSMLYHHHGINVGFTGNYQEYFSLDTAVDAVVYMMLANHLMHKLLPEATVVAEDVSGMPVLCRPVDEGGVGFDYRLAMAIPDRWIDYLKNKDDSEWSMGEIAHTLTNRRYTEKCIAYAESHDQSIVGDKTIAFLLMDKEMYTGMSDLQPASPTIDRGIALQKMIHFITMALGGDGYLNFMGNEFGHPEWIDFPREGNNWSYDKCRRQWSLVDTDHLRYKYMNAFDQAMNALDERFSFLSSSKQIVSDMNDEEKVIVFERGDLVFVFNFHPKKTYEGYKVGCDLPGKYRVALDSDALVFGGHGRVGHDVDHFTSPEGVPGVPETNFNNRPNSFKVLSPPRTCVAYYRVDEAGAGRRLHAKAETGKTSPAESIDVKASRASSKE
DKEATAGGKKGWKFARQPSDQDTK"
transit_peptide 2..190
mat_peptide 191..2467
/EC_number="2.4.1.18"
/codon start=1
/product="branching enzyme-I precursor"
polyA_signal 2734..2739BASE COUNT 719A 585C 737G 722TORIGIN
1 GCTGTGCCTC GTGTCGCCCT CTTCCTCGCC GACTCCGCTT CCGCCGCCGC GGCGCTCTCG
61 CTCGCATGCT GATCGGGCGG CACCGCCGGG GATCGCGGGT GGCGGCAATG TGCGCCTGAG
121 TGTGTTGTCT GTCCAGTGCA AGGCTCGCCG GTCAGGGGTG CGGAAGGTCA AGAGCAAATT
181 CGCCACTGCA GCTACTGTGC AAGAAGATAA AACTATGGCA ACTGCCAAAG GCGATGTCGA
241 CCATCTCCCC ATATACGACC TGGACCCCAA GCTGGAGATA TTCAAGGACC ATTTCAGGTA
301 CCGGATGAAA AGATTCCTAG AGCAGAAAGG ATCAATTGAA GAAAATGAGG GAAGTCTTGA
361 ATCTTTTTCT AAAGGCTATT TGAAATTTGG GATTAATACA AATGAGGATG GAACTGTATA
421 TCGTGAATGG GCACCTGCTG CGCAGGAGGC AGAGCTTATT GGTGACTTCA ATGACTGGAA
481 TGGTGCAAAC CATAAGATGG AGAAGGATAA ATTTGGTGTT TGGTCGATCA AAATTGACCA
541 TGTCAAAGGG AAACCTGCCA TCCCTCACAA TTCCAAGGTT AAATTTCGCT TTCTACATGG
601 TGGAGTATGG GTTGATCGTA TTCCAGCATT GATTCGTTAT GCGACTGTTG ATGCCTCTAA 661 ATTTGGAGCT CCCTATGATG GTGTTCATTG GGATCCTCCT GCTTCTGAAA GGTACACATT721 TAAGCATCCT CGGCCTTCAA AGCCTGCTGC TCCACGTATC TATGAAGCCC ATGTAGGTAT781 GAGTGGTGAA AAGCCAGCAG TAAGCACATA TAGGGAATTT GCAGACAATG TGTTGCCACG841 CATACGAGCA AATAACTACA ACACAGTTCA GTTGATGGCA GTTATGGAGC ATTCGTACTA901 TGCTTCTTTC GGGTACCATG TGACAAATTT CTTTGCGGTT AGCAGCAGAT CAGGCACACC961 AGAGGACCTC AAATATCTTG TTGATAAGGC ACACAGTTTG GGTTTGCGAG TTCTGATGGA1021 TGTTGTCCAT AGCCATGCAA GTAATAATGT CACAGATGGT TTAAATGGCT ATGATGTTGG1081 ACAAAGCACC CAAGAGTCCT ATTTTCATGC GGGAGATAGA GGTTATCATA AACTTTGGGA1141 TAGTCGGCTG TTCAACTATG CTAACTGGGA GGTATTAAGG TTTCTTCTTT CTAACCTGAG1201 ATATTGGTTG GATGAATTCA TGTTTGATGG CTTCCGATTT GATGGAGTTA CATCAATGCT1261 GTATCATCAC CATGGTATCA ATGTGGGGTT TACTGGAAAC TACCAGGAAT ATTTCAGTTT1321 GGACACAGCT GTGGATGCAG TTGTTTACAT GATGCTTGCA AACCATTTAA TGCACAAACT1381 CTTGCCAGAA GCAACTGTTG TTGCTGAAGA TGTTTCAGGC ATGCCGGTCC TTTGCCGGCC1441 AGTTGATGAA GGTGGGGTTG GGTTTGACTA TCGCCTGGCA ATGGCTATCC CTGATAGATG1501 GATTGACTAC CTGAAGAATA AAGATGACTC TGAGTGGTCG ATGGGTGAAA TAGCGCATAC1561 TTTGACTAAC AGGAGATATA CTGAAAAATG CATCGCATAT GCTGAGAGCC ATGATCAGTC1621 TATTGTTGGC GACAAAACTA TTGCATTTCT CCTGATGGAC AAGGAAATGT ACACTGGCAT1681 GTCAGACTTG CAGCCTGCTT CACCTACAAT TGATCGAGGG ATTGCACTCC AAAAGATGAT1741 TCACTTCATC ACAATGGCCC TTGGAGGTGA TGGCTACTTG AATTTTATGG GAAATGAGTT1801 TGGTCACCCA GAATGGATTG ACTTTCCAAG AGAAGGGAAC AACTGGAGCT ATGATAAATG1861 CAGACGACAG TGGAGCCTTG TGGACACTGA TCACTTGCGG TACAAGTACA TGAATGCGTT1921 TGACCAAGCG ATGAATGCGC TCGATGAGAG ATTTTCCTTC CTTTCGTCGT CAAAGCAGAT1981 CGTCAGCGAC ATGAACGATG AGGAAAAGGT TATTGTCTTT GAACGTGGAG ATTTAGTTTT2041 TGTTTTCAAT TTCCATCCCA AGAAAACTTA CGAGGGCTAC AAAGTGGGAT GCGATTTGCC1101 TGGGAAATAC AGAGTAGCCC TGGACTCTGA TGCTCTGGTC TTCGGTGGAC ATGGAAGAGT2161 TGGCCACGAC GTGGATCACT TCACGTCGCC TGAAGGGGTG CCAGGGGTGC CCGAAACGAA2221 CTTCAACAAC CGGCCGAACT CGTTCAAAGT CCTTTCTCCG CCCCGCACCT GTGTGGCTTA2281 TTACCGTGTA GACGAAGCAG GGGCTGGACG ACGTCTTCAC GCGAAAGCAG AGACAGGAAA2341 GACGTCTCCA GCAGAGAGCA TCGACGTCAA AGCTTCCAGA GCTAGTAGCA AAGAAGACAA2401 GGAGGCAACG GCTGGTGGCA AGAAGGGATG GAAGTTTGCG CGGCAGCCAT CCGATCAAGA2461 TACCAAATGA AGCCACGAGT CCTTGGTGAG GACTGGACTG GCTGCCGGCG CCCTGTTAGT2521 AGTCCTGCTC TACTGGACTA GCCGCCGCTG GCGCCCTTGG AACGGTCCTT TCCTGTAGCT2581 TGCAGGCGAC TGGTGTCTCA TCACCGAGCA GGCAGGCACT GCTTGTATAG CTTTTCTAGA2641 ATAATAATCA GGGATGGATG GATGGTGTGT ATTGGCTATC TGGCTAGACG TGCATGTGCC2701 CAGTTTGTAT GTACAGGAGC AGTTCCCGTC CAGAATAAAA AAAAACTTGT TGGGGGGTTT2761 TTC
表7玉米可溶性淀粉合成酶I基因(153bp)的转运肽
片段的编码序列和推定的氨基酸序列
[序列18和序列19]文件名:MSS1TRPT.DNA 序列:正常153BP密码子表:UNIV.TCN序列范围:1-153翻译范围:1-153★★★DNA翻译★★★1 ATG GCG ACG CCC TCG GCC GTG GGC GCC GCG TGC CTC CTC CTC GCG CGG 481 M A T P S A V G A A C L L L A R 1649 GCC GCC TGG CCG GCC GCC GTC GGC GAC CGG GCG CGC CCG CGG AGG CTC 9617 A A W P A A V G D R A R P R R L 3297 CAG CGC GTG CTG CGC CGC CGG TGC GTC GCG GAG CTG AGC AGG GAG GGG 14433 Q R V L R R R C V A E L S R E G 48145 CCC CAT ATG 15349 P H M 51GFP构建体:
1. GFP仅存在于pET-21a上:
用NdeI和XhoI消化pEXS115,并将含有SGFP编码序列的740bp的片段亚克隆到pET-(Novagen601 Science Dr.Madison WI)21a的NdeI和XhoI位点上。(见图2b,GFP-21a图谱)
2.将GFP亚克隆到读框内的全长成熟WX的5’-末端:
将源于pEXS114的含有SGFP的740bp NdeI片段亚克隆到pEXSWX的NdeI位点。(见图3a GFP-FLWX图谱)
3.将GFP亚克隆到读框内的N-末端截短了的WX的5’-末端:
将WX的N-末端截去700bp。
将源于pEXSWX的编码WX的C-末端的1kbBamHI片段亚克隆到pEXS115的BglII位点。然后将整个SGFP-截短的WX片段以NdeI-HindIII片段形式亚克隆到pET21a上。(见图3b,GFP-Bam HI WX图谱)
4.将GFP亚克隆到读框内的截短了的WX的5’-末端:将WX的N-末端截去100bp。
将源于pEXS115的含有SGFP的740bp NdeI-NcoI片段亚克隆到pEXSWX的NdeI-NcoI位点。(见图4 GFP-NcoWX图谱)
实施例3:
将质粒转化到细菌中:
制备大肠杆菌感受态细胞:
1.接种所需的大肠杆菌菌株的单菌落的2.5mlLB培养基:所选择的菌株是获自Stratagene的XLIBLUE DL21DE3;含有合适的抗生素。在37℃下,以250rpm的速度摇动,生长过夜。
2.用所述过夜培养物的1∶50的稀释液接种100ml含有合适的抗生素的LB培养基。在37℃下,以250rpm的速度摇动,生长到OD600=0.3-0.5。
3.将培养物转移到无菌离心管中,并放在冰上冷却15分钟。
4. 3,000xg(4℃)离心5分钟。
5.将沉淀重新悬浮于8ml冰冷过的转化缓冲液中。在冰上温育15分钟。
6. 3,000xg(4℃)离心5分钟。
7.将沉淀重新悬浮于8ml冰冷过的转化缓冲液2中。等分试样,在液氮中快速冷冻,并于-70℃下保存。
转化缓冲液1 转化缓冲液2
RbCl 1.2g MOPS(10mM) 0.209g
Mncl24H2O 0.99g RbCl 0.12g
乙酸钾 0.294g Cacl2H2O 1.1g
Cacl22H2O 0.15g 甘油 15g
甘油 15g 蒸馏水 100ml
蒸馏水 100ml 用NaOH调pH至6.8
用0.2M乙酸调pH至5.8 过滤消毒
过滤消毒
通过RbCl热休克法转化大肠杆菌:Hanahan,D.(1985)见DNA克隆:操作指南(Glover,D.M.著),pp.109-135,IRL出版社。
1.在冰上将1-5μl DNA与150μl大肠杆菌感受态细胞一起温育30分钟。
2.在42℃下热休克45秒。
3.紧接着在冰上放置2分钟。
4.加入600μlLB,并在37℃下温育1小时。
5.铺平板于含有合适抗生素的LB琼脂上。
该质粒将在所述细菌中表达含有绿色荧光蛋白的所述杂合多肽。
实施例4:
在大肠杆菌中表达构建体:
1.用含有感兴趣的质粒的大肠杆菌接种3mlLB。含有合适的抗生素。在37℃下,以250rpm的速度摇动,生长过夜。
2.用2ml过夜培养物接种100mlLB。含有合适的抗生素。在37℃下,以250rpm的速度摇动,生长。
3.在OD600大约为0.4-0.5的条件下于室温下放置,以200rpm的速度摇动。
4.在OD600大约为0.6-0.8的条件下用100μl 1M IPTG诱导。IPTG的终浓度为1mM。
5.在室温下,以200rpm的速度摇动,生长4-5小时。
6.通过离心收集细胞。
7.在液氮中快速冷冻,并于-70℃下保存待用。
可将细胞重新悬浮于蒸馏水中,并在UV光下(λmax=395nm)观察内源荧光。另外,可以对细胞进行超声处理,通过SDS-PAGE分离细胞提取物的等分试样,并在UV光下观察,检测GFP荧光。如果所采用的蛋白是绿色荧光蛋白,则可以在灯箱中在395nm的UV光下检测该蛋白在所述裂解材料中的存在,并可以见到标记性绿色光泽。
实施例5:
从细菌中提取质粒:
以下是可用于实施本发明的多种常用的碱性裂解质粒纯化方法之一。1.用由所述质粒之一转化过的大肠杆菌单菌落接种100-200mlLB培养基。含有合适的抗生素。在37℃下,以250rpm的速度摇动,生长过夜。2. 5,000xg(4℃)离心10分钟。3.将细胞重新悬浮于10ml水中,转移到15ml离心管中,并重复离心。4.将沉淀重新悬浮于5ml0.1MNaOH,0.5%SDS中。在冰上温育10分钟。5.加入2.5ml 3M乙酸钠(pH5.2),温和颠倒,并在冰上温育10分钟。6. 15,000-20,000xg(4℃)离心5分钟。7.用等体积的苯酚∶氯仿∶异戊醇(25∶24∶1)提取上清液。8. 6,000-10,000xg(4℃)离心10分钟。9.将水相转移到干净的离心管中,并用1倍体积的异丙醇沉淀。10. 12,000xg(4℃)离心15分钟。11.将沉淀溶解于0.5mlTE中,加入20μl10mg/mlRnase,并在37℃下温育1小时。12.用苯酚∶氯仿∶异戊醇(25∶24∶1)提取2次。13.用氯仿提取1次。14.用1倍体积的异丙醇和0.1倍体积的3M乙酸钠沉淀水相。15.用75%的乙醇洗涤沉淀1次。16.在Speed Vac中干燥沉淀,并将沉淀重新悬浮于TE中。
然后可将该质粒插入其它宿主中。
表8
源于pEXS52淀粉合成酶编码区的DNA序列和推定的氨基酸序列
[序列20和序列21]文件名:MSS1DELN.DNA 序列:正常1626BP密码子表:UNIV.TCN序列范围:1-1626翻译范围:1-1626(xi)序列描述:序列20:TGC GTC GCG GAG CTG AGC AGG GAG GAC CTC GGT CTC GAA CCT GAA GGG 48Cys Val Ala Glu Leu Ser Arg Glu Asp Leu Gly Leu Glu Pro Glu Gly
55 60 65ATT GCT GAA GGT TCC ATC GAT AAC ACA GTA GTT GTG GCA AGT GAG CAA 96Ile Ala Glu Gly Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln
70 75 80GAT TCT GAG ATT GTG GTT GGA AAG GAG CAA GCT CGA GCT AAA GTA ACA 144Asp Ser Glu Ile Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr
85 90 95CAA AGC ATT GTC TTT GTA ACC GGC GAA GCT TCT CCT TAT GCA AAG TCT 192Gln Ser Ile Val Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser100 105 110 115GGG GGT CTA GGA GAT GTT TGT GGT TCA TTG CCA GTT GCT CTT GCT GCT 240Gly Gly Leu Gly Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala
120 125 130CGT GGT CAC CGT GTG ATG GTT GTA ATG CCC AGA TAT TTA AAT GGT ACC 288Arg Gly His Arg Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr
135 140 145TCC GAT AAG AAT TAT GCA AAT GCA TTT TAC ACA GAA AAA CAC ATT CGG 336Ser Asp Lys Asn Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg
150 155 160ATT CCA TGC TTT GGC GGT GAA CAT GAA GTT ACC TTC TTC CAT GAG TAT 384Ile Pro Cys Phe Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr
165 170 175AGA GAT TCA GTT GAC TGG GTG TTT GTT GAT CAT CCC TCA TAT CAC AGA 432Arg Asp Ser Val Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg180 185 190 195CCT GGA AAT TTA TAT GGA GAT AAG TTT GGT GCT TTT GGT GAT AAT CAG 480Pro Gly Asn Leu Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln
200 205 210TTC AGA TAC ACA CTC CTT TGC TAT GCT GCA TGT GAG GCT CCT TTG ATC 528Phe Arg Tyr Thr Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile
215 220 225CTT GAA TTG GGA GGA TAT ATT TAT GGA CAG AAT TGC ATG TTT GTT GTC 576Leu Glu Leu Gly Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val
230 235 240AAT GAT TGG CAT GCC AGT CTA GTG CCA GTC CTT CTT GCT GCA AAA TAT 624Asn Asp Trp His Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr
245 250 255AGA CCA TAT GGT GTT TAT AAA GAC TCC CGC AGC ATT CTT GTA ATA CAT 672Arg Pro Tyr Gly Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile Hisu 260 265 270 275AAT TTA GCA CAT CAG GGT GTA GAG CCT GCA AGC ACA TAT CCT GAC CTT 720Asn Leu Ala His Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu
280 285 290GGG TTG CCA CCT GAA TGG TAT GGA GCT CTG GAG TGG GTA TTC CCT GAA 768Gly Leu Pro Pro Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu
295 300 305TGG GCG AGG AGG CAT GCC CTT GAC AAG GGT GAG GCA GTT AAT TTT TTG 816Trp Ala Arg Arg His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu
310 315 320AAA GGT GCA GTT GTG ACA GCA GAT CGA ATC GTG ACT GTC AGT AAG GGT 864Lys Gly Ala Val Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly
325 330 335TAT TCG TGG GAG GTC ACA ACT GCT GAA GGT GGA CAG GGC CTC AAT GAG 912Tyr Ser Trp Glu Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu340 345 350 355CTC TTA AGC TCC AGA AAG AGT GTA TTA AAC GGA ATT GTA AAT GGA ATT 960Leu Leu Ser Ser Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile
360 365 370GAC ATT AAT GAT TGG AAC CCT GCC ACA GAC AAA TGT ATC CCC TGT CAT 1008Asp Ile Asn Asp Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His
375 380 385TAT TCT GTT GAT GAC CTC TCT GGA AAG GCC AAA TGT AAA GGT GCA TTG 1056Tyr Ser Val Asp Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu
390 395 400CAG AAG GAG CTG GGT TTA CCT ATA AGG CCT GAT GTT CCT CTG ATT GGC 1104Gln Lys Glu Leu Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly
405 410 415TTT ATT GGA AGG TTG GAT TAT CAG AAA GGC ATT GAT CTC ATT CAA CTT 1152Phe Ile Gly Arg Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu420 425 430 435ATC ATA CCA GAT CTC ATG CGG GAA GAT GTT CAA TTT GTC ATG CTT GGA 1200Ile Ile Pro Asp Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly
440 445 450TCT GGT GAC CCA GAG CTT GAA GAT TGG ATG AGA TCT ACA GAG TCG ATC 1248Ser Gly Asp Pro Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile
455 460 465TTC AAG GAT AAA TTT CGT GGA TGG GTT GGA TTT AGT GTT CCA GTT TCC 1296Phe Lys Asp Lys Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser
470 475 480CAC CGA ATA ACT GCC GGC TGC GAT ATA TTG TTA ATG CCA TCC AGA TTC 1344His Arg Ile Thr Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe
485 490 495GAA CCT TGT GGT CTC AAT CAG CTA TAT GCT ATG CAG TAT GGC ACA GTT 1392Glu Pro Cys Gly Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val500 505 510 515CCT GTT GTC CAT GCA ACT GGG GGC CTT AGA GAT ACC GTG GAG AAC TTC 1440Pro Val Val His Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe
520 525 530AAC CCT TTC GGT GAG AAT GGA GAG CAG GGT ACA GGG TGG GCA TTC GCA 1488Asn Pro Phe Gly Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala
535 540 545CCC CTA ACC ACA GAA AAC ATG TTT GTG GAC ATT GCG AAC TGC AAT ATC 1536Pro Leu Thr Thr Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile
550 555 560TAC ATA CAG GGA ACA CAA GTC CTC CTG GGA AGG GCT AAT GAA GCG AGG 1584Tyr Ile Gln Gly Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg
565 570 575CAT GTC AAA AGA CTT CAC GTG GGA CCA TGC CGC TGA 1620His Val Lys Arg Leu His Val Gly Pro Cys Arg ★580 585 590
(2)序列21资料:
(i)序列特征:
(A)长度:540个氨基酸
(B)类型:氨基酸
(D)拓扑结构:线性
(ii)分子类型:蛋白
(xi)序列描述:序列21:Cys Val Ala Glu Leu Ser Arg Glu Asp Leu Gly Leu Glu Pro Glu Gly1 5 10 15Ile Ala Glu Gly Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln
20 25 30Asp Ser Glu Ile Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr
35 40 45Gln Ser Ile Val Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser
50 55 60Gly Gly Leu Gly Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala65 70 75 80Arg Gly His Arg Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr
85 90 95Ser Asp Lys Asn Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg
100 105 110Ile Pro Cys Phe Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr
115 120 125Arg Asp Ser Val Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg
130 135 140Pro Gly Asn Leu Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln145 150 155 160Phe Arg Tyr Thr Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile
165 170 175Leu Glu Leu Gly Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val
180 185 190Asn Asp Trp His Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr
195 200 205Arg Pro Tyr Gly Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile His
210 215 220Asn Leu Ala His Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu225 230 235 240Gly Leu Pro Pro Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu
245 250 255Trp Ala Arg Arg His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu
260 265 270Lys Gly Ala Val Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly
275 280 285Tyr Ser Trp Glu Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu
290 295 300Leu Leu Ser Ser Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile305 310 315 320Asp Ile Asn Asp Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His
325 330 335Tyr Ser Val Asp Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu
340 345 350Gln Lys Glu Leu Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly
355 360 365Phe Ile Gly Arg Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu
370 375 380Ile Ile Pro Asp Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly385 390 395 400Ser Gly Asp Pro Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile
405 410 415Phe Lys Asp Lys Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser
420 425 430His Arg Ile Thr Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe
435 440 445Glu Pro Cys Gly Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val
450 455 460Pro Val Val His Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe465 470 475 480Asn Pro Phe Gly Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala
485 490 495Pro Leu Thr Thr Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile
500 505 510Tyr Ile Gln Gly Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg
515 520 525His Val Lys Arg Leu His Val Gly Pro Cys Arg ★
530 535 540
实施例6:
该实施例采用了一种质粒,该质粒具有一个玉米启动子、一种玉米转运肽、一种源于淀粉合成酶基因的淀粉被囊化区、以及一个与之结合的基因片段。在图6中所示的该质粒含有表8中所示的DNA序列。
按照以下方法构建质粒pEXS52:用于构建转基因质粒的材料如下:质粒pBluescriptSK-质粒pMF6(含有nos3’终止子)质粒pHKH1(含有玉米adh1内含子)。质粒MstsI(6-4)(含有玉米stsI转运肽,用作PCTstsI转运肽产生的模板)存在于pBluescriptSK-中的质粒Mst sIII引物EXS29(GTGGATCCATGGCGACGCCCTCGGCCGTGG)[序列22]
EXS35(CTGAATTCCATATGGGGCCCCTCCCTGCTCAGCTC)[序列23]以上两种引物均被用于PCTstsI转运肽引物EXS31(CTCTGAGCTCAAGCTTGCTACTTTCTTTCCTTAATG)[序列24]EXS32(GTCTCCGCGGTGGTGTCCTTGCTTCCTAG)[序列25]两种引物均可用于PCR玉米10KD玉米醇溶蛋白启动子(杂志:基因71(359-370[1988])玉米A632基因组DNA(用作PCR玉米10KD玉米醇溶蛋白启动子的模板)。
步骤1:将玉米10KD玉米醇溶蛋白启动子克隆在pBluescriptSK-中(命名为pEXS10zp)。
1. PCR1.1Kb玉米10KD玉米醇溶蛋白启动子
引物:EXS31,EXS32
模板:玉米A632基因组DNA
2.克隆:将1.1Kb玉米10KD玉米醇溶蛋白启动子PCR产物克隆在pBluescriptSK-质粒的SacI和SacII位点上(见图7)。
步骤2:缺失pEXS10zp上的NdeI位点(被命名为pEXS10zp-NdeI)。
通过填平和平端连接pBluescriptSK中的玉米10KD玉米醇溶蛋白启动子除去NdeI。
步骤3:将玉米adh1内含子克隆到pBluescriptSK-(命名为pEXSadh1)。
玉米adh1内含子是从质粒pHKH1上的XbaI和BgmHI位点上释出的。将玉米adh1内含子(XbaI/BamHI片段)克隆到pBluescriptSK-上的XbaI和BamHI位点上(见图7)。
步骤4:将玉米10KD玉米醇溶蛋白启动子和玉米adh1内含子克隆到 pBluescriptSK-上(命名为pEXS10zp-adh1)。
玉米10KD醇溶蛋白启动子是从质粒pEXS10zp-NdeI上SaCI和SacII位点上释出的。将玉米10KD玉米醇溶蛋白启动子(SaCI/SacII片段)克隆到质粒pEXSadh1(含有玉米adh1内含子)的SacI和SacII位点上(见图7)。
步骤5:将玉米nos3’终止子克隆到质粒pEXSadh1上(命名为pEXSadh1-nos3’)。
玉米nos3’终止子是从质粒pMF6上的EcoRI和HindIII位点上释出的。将玉米nos3’终止子(EcoRI/HindIII片段)克隆到质粒pEXSadh1上的EcoRI和HindIII上(见图7)。
步骤6:将玉米nos3’终止子克隆到质粒pEXS10zp-adh1上(命名为pEXS10zp-adh1-nos3’)。
玉米nos3’终止子是从质粒pEXSadh1-nos3’上的EcoRI和ApeI位点上释出的。将玉米nos3’终止子(EcoRI/ApaI片段)克隆到质粒pEXS10zp-adh1上的EcoRI和ApaI位点上(见图7)。
步骤7:将玉米STSI转运肽克隆到质粒pEXS10zp-adh1-nos3’上(命名为pEXS33)。
1. PCR150bp玉米STSI转运肽引物:EXS29,EXS35]
模板:MSTSI(6-4)质粒
2.将150bp玉米STSI转运肽PCR产物克隆到质粒pEXS10zp-adh1-nos3’上的EcoRI和BamHI位点上(见图7)。
步骤8:对pEXS33上的玉米STSI转运肽进行定点诱变(命名为pEXS33(m))。
在质粒pEXS33上的玉米STSI转运肽上存在一个突变(终止密码子)。进行定点诱变是为了将终止密码子改变成非终止密码子。新的质粒(含有玉米10KD玉米醇溶蛋白启动子、玉米STSI转运肽、玉米adh1内含子、玉米nos3’终止子)被命名为pEXS33(m)。
步骤9:使pEXS33(m)上的NotI位点缺失(命名为pEXS50)。
通过NotI补平和平端连接除去pEXS33上的NotI位点,以形成pEXS50(见图8)。
步骤10:使pEXS33(m)上的玉米adh1内含子缺失(命名为pEXS60上)。
通过NotI/BamHI消化,用Klenow片段补平和平端连接除去玉米adh1内含子,以生成pEXS60(见图9)。
步骤11:将玉米STSIII克隆到pEXS50、pEXS60上。
玉米STSIII是从质粒pBluescriptSK-上的NdeI和EcoRI位点上释出的。将玉米STSIII(NdeI-EcoRI片段分别克隆到pEXS50、pEXS60上,命名为pEXS51、pEXS61(分别参见图8和图9) 。
步骤12:将表8所示基因克隆到pEXS51的NdeI/NotI位点上,以生成pEXS52。可以通过将其它基因(STSI、II、WX、glgA、glgB、glgC、BEI、BEII等)克隆到质粒pEXS51、pEXS61的NdeI/NotI位点上制备其它类似的质粒。将质粒pEXS52转入稻中。对用pEXS52转化过的再生稻植株进行标记,并将其放入品红箱中。
从所述品红箱中选择每个系的两个姊妹株,并将其转入装有土壤混合物(表土与泥炭-蛭石50/50混合物)的2.5英寸的盆中。将所述盆放在装有半英寸高水的水族箱中(鱼箱)。盖住该箱的上部以保持高湿度(在其上面设置一些孔,以便散热)。用一个温度计监测其温度。将该鱼箱放在荧光灯下。在第一周不对所述植株施肥。光照周期为上午6点-下午8点,最少有14小时的光照。夜间温度最低为68°F,日间温度为80-90°F。在所述鱼箱下面使用一个加热垫,以便在需要时帮助根系生长。让所述植物在上述条件下保持一周。(注:所述姊妹株应为低光照强度而长高)。
在上述第一周之后,打开所述水族箱的顶部,并将稻转化体转移到生长箱中,在高湿度和高光照强度下生长三周。
另外,可以在温室中用水混合物保持高湿度,让植株生长三周。然后将该植株转移到装有土壤混合物(表土与泥炭-蛭石50/50混合物)的6英寸的盆中(最小为5英寸盆)。将所述盆放入装有半英寸水的托盘中。用15-16-17(N-K-P)对所述植株施肥(250ppm),每周一次,或根据植株的需要通过观察其外观进行施肥。让所述植株保持14小时光照(最低),上午6点-下午8点的高光照强度,温度为85-90°F/70°F日间/夜间。
让所述植株产生稻粒,并收获这些稻粒。从所收获的种子中提取淀粉,并分析这些种子的淀粉中连接氨基酸C,V,A,E,L,S,R[序列27]的存在。
实施例7:
用于植物的SER载体:
图6所示质粒适用于单子叶植物,即玉米。质粒pEXS52(图6)具有一个启动子,一个转运肽(源于玉米),和一个编码氨基酸序列CVAELSRE[序列27]的连接基因片段(TGC GTC GCG GAGCTG AGC AGG GAG)[序列26]。
该基因片段天然存在于接近玉米可溶性淀粉合成酶(MSTSI)基因的N-末端处。如表8所示,源于淀粉合成酶的SER始于氨基酸292位左右。优选将该载体转入玉米宿主中。该转运多肽适用于玉米,因此玉米是优选的宿主。很显然,如果必要的话,所述转运肽和启动子可加以改变,以适应所希望的宿主植物。在通过“whiskers”技术(美国专利US5,302,523和US5,464,765)转化以后,通过本领域已知方法再生转化的宿主细胞,对转化株进行授粉,并收集所得到的种子,分析所述肽在淀粉和淀粉粒中的存在。
可将以下优选的基因用于玉米中以改善其种子:肌醇六磷酸酶基因,促生长素基因,以下链状氨基酸:AUG AUG AUG AUG AUG AUGAUG AUG[序列 28],和/或AAG AAG AAG AAG AAG AAG AAG AAG AAGAAG AAG AAG[序列29];和/或AAA AAA AAA AAA AAA AAA[序列30];或编码一种链状赖氨酸的密码子的组合或编码赖氨酸密码子和甲硫氨酸密码子的组合或任意两种或三种此类氨基酸的组合。所述链的长度不宜过长,不过,链的长度似乎并不重要。这样,所述氨基酸就被被囊化于淀粉粒中或结合于淀粉粒中,形成所述植物宿主的含有淀粉的部分。
该质粒可以被转入诸如稻、小麦、大麦、燕麦、高粱、或粟的其它禾本科植物中,只需对该质粒作很小的改进或不作改进。所述启动子可以是waxy基因启动子,其序列已被公开,或本领域已知的其它玉米醇溶蛋白启动子。
另外,不用做太多的试验即可将上述质粒转入诸如马铃薯、甘薯、芋、大薯、荷、木薯、花生、豌豆、大豆、菜豆、或鹰嘴豆的双子叶植物中。可以对启动子进行选择,以便将特定双子叶植物或块茎的淀粉储存部位定位,例如可将patatin启动子用于马铃薯块茎。
转化单子叶植物和双子叶植物的各种方法在本领域中是公知的,而且转化基因的方法对本发明来说并不重要。可以用An等的冻-融法将所述质粒导入根癌农杆菌中(Agrobacteriumtumefaciens)(1988,二元载体,见植物分子生物学手册A3,S.D.Gelvin R.A.Schilperoot著,Dordrecht,荷兰:Kluwer学术出版社,pp.1-19)。在Edwards等的著述中披露了制备含有所述构建体的农杆菌接种物的方法,并接种植物材料,再生芽,以及使芽长根(“源于马铃薯的新型淀粉合成酶的生物化学和分子鉴定”,植物杂志8,283-294(1995))。
在多种不同的基因中存在着多种被囊化区。尽管优选将所述蛋白被囊化于淀粉粒中(粒状被囊化作用),但本发明所述的“被囊化”一词的含义还包括被囊化于非粒状淀粉中。以下类型的基因可用于这一目的。糖原合成酶的淀粉被囊化区的用途:
大肠杆菌糖原合成酶不是一种大的蛋白:其结构基因的长度为1431bp,编码一种具有477个氨基酸的蛋白,其估计分子量为49,000。众所周知的是,在将细菌基因插入植物基因组时会出现密码子使用问题,但是,这一问题对于大肠杆菌基因来说不像源于诸如芽孢杆菌的其它细菌的基因那么严重。源于大肠杆菌的糖原合成酶的密码子使用特征与玉米基因具有更多的不同之处,不过,优选用已知方法对位于其翻译起始点的序列加以改变,以便与植物共有序列更加一致:
glgA GATAATGCAG[序列31]
共有序列AACAATGGCT[序列32]可溶性淀粉合成酶的淀粉被囊化区的用途:
在上文的背景技术部分披露了植物可溶性淀粉合成酶的cDNA克隆,可将这些克隆用于本发明中。根据本发明,编码上述任何SSTS蛋白的基因均可用于构建体中。分支酶的淀粉被囊化区的用途:
可将在上文的背景技术部分所披露的植物、细菌和动物分支酶的cDAN克隆用于本发明中。能将直链淀粉转化成支链淀粉(将一段1,4D葡聚糖链转移到一个类似的葡聚糖链的伯位羟基上)的分支酶[1,4D萄聚糖:1,4D葡聚糖6D(1,4D葡聚糖)转移酶(E.C.2.4.1.18)]有时被称为Q-酶。
Baba等研究了玉米分支酶I的序列,(1991,BBRC.181:87-94)。Fisher等研究了源于玉米胚乳的淀粉分支酶II(1993,植物生理学,102:1045-1046)。所述BE基因构建体可能需要由一种造粉体转运肽来确保其在造粉体中的正确定位。根据本发明,编码上述GBSTS蛋白的分支酶的任一种的基因均可用构建体中。
颗粒结合淀粉合成酶的淀粉结合域的用途:植物颗粒结合淀粉合成酶的cDNA的用途披露于下列文献中:Shure等(1983)细胞35:225-233,Visser等(1989)植物科学64(2):185-192。Visser等还披露了用反义构建体在马铃薯中抑制编码颗粒结合淀粉合成酶的基因的表达(1991,分子基因遗传学225(2):289-296;1994,植物细胞6:43-52)。Shimada等在稻中证实了反义现象(1993,理论应用遗传学86:665-672)。Van der Leij等证实,在用野生型waxy马铃薯基因转化以后可在低支链淀粉马铃薯中恢复直链淀粉的合成(1991,理论应用遗传学82:289-295) 。
源于诸如玉米、稻、小麦、马铃薯、木薯、豌豆、或大麦的颗粒淀粉合成酶的氨基酸序列和核苷酸序列是众所周知的。根据本发明,编码上述任一种GBSTS蛋白的基因均可用于构建体中。构建植物转化载体:
用于本发明方法中的植物转化载体可以用标准技术构建。转运肽序列的用途:
某些基因构建体需要由造粉体转运肽来确保在造粉体中的正确定位。据信,叶绿体转运肽具有相似的序列(Heijne等披露了叶绿体转运肽的数据,见:1991植物分子生物学报导,9(2:104-126)。可用于本发明的其它转运肽为ADPG焦磷酸化酶(1991植物分子生物学报导,9:104-126),小亚基RUBISCO,乙酰乳酸合成酶、甘油醛三磷酸脱氢酶和亚硝酸还原酶。
源于许多基因型的小亚基RUBISCO转运肽的共有序列的序列如下:MASSMLSSAAVATRTNPAQASMVAPFTGLKSAAFPVSRKQNLDITSIASNGGRVQC[序列33]玉米小亚基RUBISCO具有以下序列:MAPTVMMASSATATRTNPAQASAVAPFQGLKSTASLPVARRSSRSLGNVASNGGRIRC[序列34]
源于玉米叶片的三磷酸甘油醛脱氢酶的转动肽具有如下序列:MAQILAPSTQWQMRITKTSPCATPITSKMWSSLVMKQTKKVAHSAKFRVMAVNSENGT[序列35]。
玉米胚乳结合淀粉合成酶的转运肽序列具有以下序列:MAALATSQLVATRAGHGVPDASTFRRGAAQGLRGARASAAADTLSMRTSARAAPRHQQQARRGGRFPFPSLVVC[序列36]
玉米胚乳可溶性淀粉合成酶的转运肽序列具有以下序列:MATPSAVGAACLLLARXAWPAAVGDRARPRRLQRVLRRR[序列37]
通过工程方法将新的氨基酸或肽插入淀粉被囊化蛋白:
可以用本领域技术人员公知的方法插入新的氨基酸组合用于本发明所述的淀粉结合蛋白的修饰。例如,可以对淀粉结合蛋白的序列进行修饰,以表达高于正常水平的赖氨酸、甲硫氨酸或色氨酸。这种水平高于天然水平是有用的,而且这种蛋白可使诸如禾本科植物的作物的营养得到加强。
除了改变氨基酸的平衡以外,还可以对淀粉结合蛋白进行工程操作,以便将有价值的肽插入所述淀粉结合蛋白中。将所述有效负荷多肽结合在淀粉结合蛋白的N-末端,这是一种将肽片段添加上去但仍能维持其淀粉结合能力的公知方法。还可以通过将特定的蛋白酶裂解位点插入所述有效负荷多肽与所述淀粉被囊化区的结合位点对其作进一步的改进。本领域技术人员熟知的是,蛋白酶对于不同的氨基酸键具有优选的专一性。这种专一性可被用于提供一种载体,以便将有价值的肽输送到动物和人的消化道的不同部位。
在本发明的另一种实施方案中,在对所述淀粉粒进行纯化和加工以后,可以释放出所述有效负荷多肽。众所周知,通过解淀粉和/或凝胶化方法可以释放出与淀粉粒结合的蛋白或使其变得可用于蛋白水解作用。因此,有可能从淀粉粒中回收到有商业价值量的蛋白和肽。
在本发明的又一种实施方案中,可以用多种不同方法处理所述淀粉粒,以提供改变该淀粉的可消化性的方法。采用这种方法,有可能改变结合在淀粉粒中的蛋白、肽或氨基酸的生物可利用性。虽然为清楚和理解起见已通过说明和举例的方式对上述发明作了详细说明,但对本领域普通技术人员来说,显而易见的是,通过阅读本发明可以在不超出所附权利要求的构思或范围的前提下对本发明作出某些改变和改进。
序列表(1)一般资料:
(i)申请人:Keeling,Peter
Guan,Hanping
(ii)发明名称:淀粉被囊化
(iii)序列数:37
(iv)通信地址:
(A)收件人:Greenlee,Winner和Sullivan,P.C.
(B)街道:5370 Manhattan Circle
(C)城市:Boulder
(D)州:CO
(E)国家:美国
(F)邮编:80303
(v)计算机可读形式:
(A)媒体类型:软盘
(B)计算机:IBM PC兼容
(C)操作系统:C-DOS/MS-DOS
(D)软件:PatentIn Release#1.0,Version#1.30
(vi)当前申请数据:
(A)申请号:US
(B)申请日:30-9-1997
(C)分类号:
(vii)在先申请数据:
(A)申请号:US 60/026,855
(B)申请日:30-9-1996
(viii)律师/代理人资料:
(A)姓名:Winner,Ellen P
(B)注册号:28,547
(C)资料/文件号:89-97
(ix)通信资料:
(A)电话:(303)499-8080
(B)传真:(303)499-8089(2)序列1资料:
(i)序列特征:
(A)长度:31个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:线形
(ii)分子类型:其它核酸
(A)说明:/desc=“寡核苷酸”
(iii)假说:无
(xi)序列描述:序列1:GACTAGTCAT ATGGTGAGCA AGGGCGAGGA G(2)序列2资料:
(i)序列特征:
(A)长度:36个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:线形
(ii)分子类型:其它核酸
(A)说明:/desc=“寡核苷酸”
(iii)假说:无
(xi)序列描述:序列2:CTAGATCTTC ATATGCTTGT ACAGCTCGTC CATGCC(2)序列3资料:
(i)序列特征:
(A)长度:39个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:线形
(ii)分子类型:其它核酸
(A)说明:/desc=“寡核苷酸”
(iii)假说:无
(xi)序列描述:序列3:CTAGATCTTG GCCATGGCCT GGTACAGCTC GTCCATGCC(2)序列4资料:
(i)序列特征:
(A)长度:4800个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:DNA(基因组)
(iii)假说:无
(vi)来源:
(A)生物:玉米
(iv)特征:
(A)名称/键:CDS
(B)位置:结合(1449..1553,1685..1765,1860..1958,2055..2144,2226..2289,2413..2513,2651..2760,2858..3101,3212..3394,3490..3681,3793..3879,3977..4105,4227..4343)
(xi)序列描述:序列4:CAGCGACCTA TTACACAGCC CGCTCGGGCC CGCGACGTCG GGACACATCT TCTTCCCCCT 60TTTGGTGAAG CTCTGCTCGC AGCTGTCCGG CTCCTTGGAC GTTCGTGTGG CAGATTCATC 120TGTTGTCTCG TCTCCTGTGC TTCCTGGGTA GCTTGTGTAG TGGAGCTGAC ATGGTCTGAG 180CAGGCTTAAA ATTTGCTCGT AGACGAGGAG TACCAGCACA GCACGTTGCG GATTTCTCTG 240CCTGTGAAGT GCAACGTCTA GGATTGTCAC ACGCCTTGGT CGCGTCGCGT CGCGTCGCGT 300CGATGCGGTG GTGAGCAGAG CAGCAACAGC TGGGCGGCCC AACGTTGGCT TCCGTGTCTT 360CGTCGTACGT ACGCGCGCGC CGGGGACACG CAGCAGAGAG CGGAGAGCGA GCCGTGCACG 420GGGAGGTGGT GTGGAAGTGG AGCCGCGCGC CCGGCCGCCC GCGCCCGGTG GGCAACCCAA 480AAGTACCCAC GACAAGCGAA GGCGCCAAAG CGATCCAAGC TCCGGAACGC AACAGCATGC 540GTCGCGTCGG AGAGCCAGCC ACAAGCAGCC GAGAACCGAA CCGGTGGGCG ACGCGTCATG 600GGACGGACGC GGGCGACGCT TCCAAACGGG CCACGTACGC CGGCGTGTGC GTGCGTGCAG 660ACGACAAGCC AAGGCGAGGC AGCCCCCGAT CGGGAAAGCG TTTTGGGCGC GAGCGCTGGC 720GTGCGGGTCA GTCGCTGGTG CGCAGTGCCG GGGGGAACGG GTATCGTGGG GGGCGCGGGC 780GGAGGAGAGC GTGGCGAGGG CCGAGAGCAG CGCGCGGCCG GGTCACGCAA CGCGCCCCAC 840GTACTGCCCT CCCCCTCCGC GCGCGCTAGA AATACCGAGG CCTGGACCGG GGGGGGGCCC 900CGTCACATCC ATCCATCGAC CGATCGATCG CCACAGCCAA CACCACCCGC CGAGGCGACG 960CGACAGCCGC CAGGAGGAAG GAATAAACTC ACTGCCAGCC AGTGAAGGGG GAGAAGTGTA 1020CTGCTCCGTC GACCAGTGCG CGCACCGCCC GGCAGGGCTG CTCATCTCGT CGACGACCAG 1080GTTCTGTTCC GTTCCGATCC GATCCGATCC TGTCCTTGAG TTTCGTCCAG ATCCTGGCGC 1140GTATCTGCGT GTTTGATGAT CCAGGTTCTT CGAACCTAAA TCTGTCCGTG CACACGTCTT 1200TTCTCTCTCT CCTACGCAGT GGATTAATCG GCATGGCGGC TCTGGCCACG TCGCAGCTCG 1260TCGCAACGCG CGCCGGCCTG GGCGTCCCGG ACGCGTCCAC GTTCCGCCGC GGCGCCGCGC 1320AGGGCCTGAG GGGGGCCCGG GCGTCGGCGG CGGCGGACAC GCTCAGCATG CGGACCAGCG 1380CGCGCGCGGC GCCCAGGCAC CAGCAGCAGG CGCGCCGCGG GGGCAGGTTC CCGTCGCTCG 1440TCGTGTGC GCC AGC GCC GGC ATG AAC GTC GTC TTC GTC GGC GCC GAG ATG 1490
Ala Ser Ala Gly Met Asn Val Val Phe Val Gly Ala Glu Met
1 5 10GCG CCG TGG AGC AAG ACC GGC GGC CTC GGC GAC GTC CTC GGC GGC CTG 1538Ala Pro Trp Ser Lys Thr Gly Gly Leu Gly Asp Val Leu Gly Gly Leu15 20 25 30CCG CCG GCC ATG GCC GTAAGCGCGC GCACCGAGAC ATGCATCCGT TGGATCGC GT 1593Pro Pro Ala Met Ala
35CTTCTTCGTG CTCTTGCCGC GTGCATGATG CATGTGTTTC CTCCTGGCTT GTGTTCGTGT 1653ATGTGACGTG TTTGTTCGGG CATGCATGCA G GCG AAC GGG CAC CGT GTC ATG 1705
Ala Asn Gly His Arg Val Met
40GTC GTC TCT CCC CGC TAC GAC CAG TAC AAG GAC GCC TGG GAC ACC AGC 1753Val Val Ser Pro Arg Tyr Asp Gln Tyr Lys Asp Ala Trp Asp Thr Ser
45 50 55GTC GTG TCC GAG GTACGGCCAC CGAGACCAGA TTCAGATCAC AGTCACACAC 1805Val Val Ser Glu
60ACCGTCATAT GAACCTTTCT CTGCTCTGAT GCCTGCAACT GCAAATGCAT GCAG ATC 1862
IleAAG ATG GGA GAC GGG TAC GAG ACG GTC AGG TTC TTC CAC TGC TAC AAG 1910Lys Met Gly Asp Gly Tyr Glu Thr Val Arg Phe Phe His Cys Tyr Lys
65 70 75CGC GGA GTG GAC CGC GTG TTC GTT GAC CAC CCA CTG TTC CTG GAG AGG 1958Arg Gly Val Asp Arg Val Phe Val Asp His Pro Leu Phe Leu Glu Arg80 85 90 95GTGAGACGAG ATCTGATCAC TCGATACGCA ATTACCACCC CATTGTAAGC AGTTACAGTG 2018AGCTTTTTTT CCCCCCGGCC TGGTCGCTGG TTTCAG GTT TGG GGA AAG ACC GAG 2072
Val Trp Gly Lys Thr Glu
100GAG AAG ATC TAC GGG CCT GTC GCT GGA ACG GAC TAC AGG GAC AAC CAG 2120Glu Lys Ile Tyr Gly Pro Val Ala Gly Thr Asp Tyr Arg Asp Asn Gln
105 110 115CTG CGG TTC AGC CTG CTA TGC CAG GTCAGGATGG CTTGGTACTA CAACTTCATA 2174Leu Arg Phe Ser Leu Leu Cys Gln
120 125TCATCTGTAT GCAGCAGTAT ACACTGATGA GAAATGCATG CTGTTCTGCA G GCA GCA 2231
Ala AlaCTT GAA GCT CCA AGG ATC CTG AGC CTC AAC AAC AAC CCA TAC TTC TCC 2279Leu Glu Ala Pro Arg Ile Leu Ser Leu Asn Asn Asn Pro Tyr Phe Ser
130 135 140GGA CCA TAC G GTAAGAGTTG CAGTCTTCGT ATATATATCT GTTGAGCTCG 2329Gly Pro Tyr
145AGAATCTTCA CAGGAAGCGG CCCATCAGAC GGACTGTCAT TTTACACTGA CTACTGCTGC 2389TGCTCTTCGT CCATCCATAC AAG GG GAG GAC GTC GTG TTC GTC TGC AAC 2438
Gly Glu Asp Val Val Phe Val Cys Asn
150 155GAC TGG CAC ACC GGC CCT CTC TCG TGC TAC CTC AAG AGC AAC TAC CAG 2486Asp Trp His Thr Gly Pro Leu Ser Cys Tyr Leu Lys Ser Asn Tyr Gln
160 165 170TCC CAC GGC ATC TAC AGG GAC GCA AAG GTTGCCTTCT CTGAACTGAA 2533Ser His Gly Ile Tyr Arg Asp Ala Lys
175 180CAACGCCGTT TTCGTTCTCC ATGCTCGTAT ATACCTCGTC TGGTAGTGGT GGTGCTTCTC 2593TGAGAAACTA ACTGAAACTG ACTGCATGTC TGTCTGACCA TCTTCACGTA CTACCAG 2650ACC GCT TTC TGC ATC CAC AAC ATC TCC TAC CAG GGC CGG TTC GCC TTC 2698Thr Ala Phe Cys Ile His Asn Ile Ser Tyr Gln Gly Arg Phe Ala Phe
185 190 195TCC GAC TAC CCG GAG CTG AAC CTC CCG GAG AGA TTC AAG TCG TCC TTC 2746Ser Asp Tyr Pro Glu Leu Asn Leu Pro Glu Arg Phe Lys Ser Ser Phe
200 205 210GAT TTC ATC GAC GG GTCTGTTTTC CTGCGTGCAT GTGAACATTC ATGAATGGTA 2800Asp Phe Ile Asp Gly
215ACCCACAACT GTTCGCGTCC TGCTGGTTCA TTATCTGACC TGATTGCATT ATTGCAG C 2858TAC GAG AAG CCC GTG GAA GGC CGG AAG ATC AAC TGG ATG AAG GCC GGG 2906Tyr Glu Lys Pro Val Glu Gly Arg Lys Ile Asn Trp Met Lys Ala Gly
220 225 230ATC CTC GAG GCC GAC AGG GTC CTC ACC GTC AGC CCC TAC TAC GCC GAG 2954Ile Leu Glu Ala Asp Arg Val Leu Thr Val Ser Pro Tyr Tyr Ala Glu
235 240 245GAG CTC ATC TCC GGC ATC GCC AGG GGC TGC GAG CTC GAC AAC ATC ATG 3002Glu Leu Ile Ser Gly Ile Ala Arg Gly Cys Glu Leu Asp Asn Ile Met250 255 260 265CGC CTC ACC GGC ATC ACC GGC ATC GTC AAC GGC ATG GAC GTC AGC GAG 3050Arg Leu Thr Gly Ile Thr Gly Ile Val Asn Gly Met Asp Val Ser Glu
270 275 280TGG GAC CCC AGC AGG GAC AAG TAC ATC GCC GTG AAG TAC GAC GTG TCG 3098Trp Asp Pro Ser Arg Asp Lys Tyr Ile Ala Val Lys Tyr Asp Val Ser
285 290 295ACG GTGAGCTGGC TAGCTCTGAT TCTGCTGCCT GGTCCTCCTG CTCATCATGC 3151ThrTGGTTCGGTA CTGACGCGGC AAGTGTACGT ACGTGCGTGC GACGGTGGTG TCCGGTTCAG 3211GCC GTG GAG GCC AAG GCG CTG AAC AAG GAG GCG CTG CAG GCG GAG GTC 3259Ala Val Glu Ala Lys Ala Leu Asn Lys Glu Ala Leu Gln Ala Glu Val
300 305 310GGG CTC CCG GTG GAC CGG AAC ATC CCG CTG GTG GCG TTC ATC GGC AGG 3307Gly Leu Pro Val Asp Arg Asn Ile Pro Leu Val Ala Phe Ile Gly Arg315 320 325 330CTG GAA GAG CAG AAG GGC CCC GAC GTC ATG GCG GCC GCC ATC CCG CAG 3355Leu Glu Glu Gln Lys Gly Pro Asp Val Met Ala Ala Ala Ile Pro Gln
335 340 345CTC ATG GAG ATG GTG GAG GAC GTG CAG ATC GTT CTG CTG GTACGTGTGC 3404Leu Met Glu Met Val Glu Asp Val Gln Ile Val Leu Leu
350 355GCCGGCCGCC ACCCGGCTAC TACATGCGTG TATCGTTCGT TCTACTGGAA CATGCGTGTG 3464AGCAACGCGA TGGATAATGC TGCAG GGC ACG GGC AAG AAG AAG TTC GAG CGC 3516
Gly Thr Gly Lys Lys Lys Phe Glu Arg
360 365ATG CTC ATG AGC GCC GAG GAG AAG TTC CCA GGC AAG GTG CCC GCC GTG 3564Met Leu Met Ser Ala Glu Glu Lys Phe Pro Gly Lys Val Arg Ala Val
370 375 380GTC AAG TTC AAC GCG GCG CTG GCG CAC CAC ATC ATG GCC GGC GCC GAC 3612Val Lys Phe Asn Ala Ala Leu Ala His His Ile Met Ala Gly Ala Asp385 390 395 400GTG CTC GCC GTC ACC AGC CGC TTC GAG CCC TGC GGC CTC ATC CAG CTG 3660Val Leu Ala Val Thr Ser Arg Phe Glu Pro Cys Gly Leu Ile Gln Leu
405 410 415CAG GGG ATG CGA TAC GGA ACG GTACGAGAGA AAAAAAAAAT CCTGAATCCT 3711Gln Gly Met Arg Tyr Gly Thr
420GACGAGAGGG ACAGAGACAG ATTATGAATG CTTCATCGAT TTGAATTGAT TGATCGATGT 3771CTCCCGCTGC GACTCTTGCA G CCC TGC GCC TGC GCG TCC ACC GGT GGA CTC 3822
Pro Cys Ala Cys Ala Ser Thr Gly Gly Leu
425 430GTC GAC ACC ATC ATC GAA GGC AAG ACC GGG TTC CAC ATG GGC CGC CTC 3870Val Asp Thr Ile Ile Glu Gly Lys Thr Gly Phe His Met Gly Arg Leu
435 440 445AGC GTC GAC GTAAGCCTAG CTCTGCCATG TTCTTTCTTC TTTCTTTCTG 3919Ser Val Asp450TATGTATGTA TGAATCAGCA CCGCCGTTCT TGTTTCGTCG TCGTCCTCTC TTCCCAG 3976TGT AAC GTC GTG GAG CCG GCG GAC GTC AAG AAG GTG GCC ACC ACA TTG 4024Cys Asn Val Val Glu Pro Ala Asp Val Lys Lys Val Ala Thr Thr Leu
455 460 465CAG CGC GCC ATC AAG GTG GTC GGC ACG CCG GCG TAC GAG GAG ATG GTG 4072Gln Arg Ala Ile Lys Val Val Gly Thr Pro Ala Tyr Glu Glu Met Val
470 475 480AGG AAC TGC ATG ATC CAG GAT CTC TCC TGG AAG GTACGTACGC CCGCCCCGCC 4125Arg Asn Cys Met Ile Gln Asp Leu Ser Trp Lys485 490 495CCGCCCCGCC AGAGCAGAGC GCCAAGATCG ACCGATCGAC CGACCACACG TACGCGCCTC 4185GCTCCTGTCG CTGACCGTGG TTTAATTTGC GAAATGCGCA G GGC CCT GCC AAG 4238
Gly Pro Ala LysAAC TGG GAG AAC GTG CTG CTC AGC CTC GGG GTC GCC GGC GGC GAG CCA 4286Asn Trp Glu Asn Val Leu Leu Ser Leu Gly Val Ala Gly Gly Glu Pro500 505 510 515GGG GTC GAA GGC GAG GAG ATC GCG CCG CTC GCC AAG GAG AAC GTG GCC 4334Gly Val Glu Gly Glu Glu Ile Ala Pro Leu Ala Lys Glu Asn Val Ala
520 525 530GCG CCC TGA AGAGTTCGGC CTGCAGGGCC CCTGATCTCG CGCGTGGTGC 4383Ala Pro ★AAAGATGTTG GGACATCTTC TTATATATGC TGTTTCGTTT ATGTGATATG GACAAGTATG 4443TGTAGCTGCT TGCTTGTGCT AGTGTAATGT AGTGTAGTGG TCGCCAGTGG CACAACCTAA 4503TAAGCGCATG AACTAATTGC TTGCGTGTGT AGTTAAGTAC CGATCGGTAA TTTTATATTG 4563CGAGTAAATA AATGGACCTG TAGTGGTGGA GTAAATAATC CCTGCTGTTC GGTGTTCTTA 4623TCGCTCCTCG TATAGATATT ATATAGAGTA CATTTTTCTC TCTCTGAATC CTACGTTTGT 4683GAAATTTCTA TATCATTACT GTAAAATTTC TGCGTTCCAA AAGAGACCAT AGCCTATCTT 4743TGGCCCTGTT TGTTTCGGCT TCTGGCAGCT TCTGGCCACC AAAAGCTGCT GCGGACT 4800(2)序列5资料:
(i)序列特征:
(A)长度:534氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列5:Ala Ser Ala Gly Met Asn Va1 Val Phe Val Gly Ala Glu Met Ala Pro1 5 10 15Trp Ser Lys Thr Gly Gly Leu Gly Asp Val Leu Gly Gly Leu Pro Pro
20 25 30Ala Met Ala Ala Asn Gly His Arg Val Met Val Val Ser Pro Arg Tyr
35 40 45Asp Gln Tyr Lys Asp Ala Trp Asp Thr Ser Val Val Ser Glu Ile Lys
50 55 60Met Gly Asp Gly Tyr Glu Thr Val Arg Phe Phe His Cys Tyr Lys Arg65 70 75 80Gly Val Asp Arg Val Phe Val Asp His Pro Leu Phe Leu Glu Arg Val
85 90 95Trp Gly Lys Thr Glu Glu Lys Ile Tyr Gly Pro Val Ala Gly Thr Asp
100 105 110Tyr Arg Asp Asn Gln Leu Arg Phe Ser Leu Leu Cys Gln Ala Ala Leu
115 120 125Glu Ala Pro Arg Ile Leu Ser Leu Asn Asn Asn Pro Tyr Phe Ser Gly
130 135 140Pro Tyr Gly Glu Asp Val Val Phe Val Cys Asn Asp Trp His Thr Gly145 150 155 160Pro Leu Ser Cys Tyr Leu Lys Ser Asn Tyr Gln Ser His Gly Ile Tyr
165 170 175Arg Asp Ala Lys Thr Ala Phe Cys Ile His Asn Ile Ser Tyr Gln Gly
180 185 190Arg Phe Ala Phe Ser Asp Tyr Pro Glu Leu Asn Leu Pro Glu Arg Phe
195 200 205Lys Ser Ser Phe Asp Phe Ile Asp Gly Tyr Glu Lys Pro Val Glu Gly
210 215 220Arg Lys Ile Asn Trp Met Lys Ala Gly Ile Leu Glu Ala Asp Arg Val225 230 235 240Leu Thr Val Ser Pro Tyr Tyr Ala Glu Glu Leu Ile Ser Gly Ile Ala
245 250 255Arg Gly Cys Glu Leu Asp Asn Ile Met Arg Leu Thr Gly Ile Thr Gly
260 265 270Ile Val Asn Gly Met Asp Val Ser Glu Trp Asp Pro Ser Arg Asp Lys
275 280 285Tyr Ile Ala Val Lys Tyr Asp Val Ser Thr Ala Val Glu Ala Lys Ala
290 295 300Leu Asn Lys Glu Ala Leu Gln Ala Glu Val Gly Leu Pro Val Asp Arg305 310 315 320Asn Ile Pro Leu Val Ala Phe Ile Gly Arg Leu Glu Glu Gln Lys Gly
325 330 335Pro Asp Val Met Ala Ala Ala Ile Pro Gln Leu Met Glu Met Val Glu
340 345 350Asp Val Gln Ile Val Leu Leu Gly Thr Gly Lys Lys Lys Phe Glu Arg
355 360 365Met Leu Met Ser Ala Glu Glu Lys Phe Pro Gly Lys Val Arg Ala Val
370 375 380Val Lys Phe Asn Ala Ala Leu Ala His is Ile Met Ala Gly Ala Asp385 390 395 400Val Leu Ala Val Thr Ser Arg Phe Glu Pro Cys Gly Leu Ile Gln Leu
405 410 415Gln Gly Met Arg Tyr Gly Thr Pro Cys Ala Cys Ala Ser Thr Gly Gly
420 425 430Leu Val Asp Thr Ile Ile Glu Gly Lys Thr Gly Phe His Met Gly Arg
435 440 445Leu Ser Val Asp Cys Asn Val Val Glu Pro Ala Asp Val Lys Lys Val
450 455 460Ala Thr Thr Leu Gln Arg Ala Ile Lys Val Val Gly Thr Pro Ala Tyr465 470 475 480Glu Glu Met Val Arg Asn Cys Met Ile Gln Asp Leu Ser Trp Lys Gly
485 490 495Pro Ala Lys Asn Trp Glu Asn Val Leu Leu Ser Leu Gly Val Ala Gly
500 505 510Gly Glu Pro Gly Val Glu Gly Glu Glu Ile Ala Pro Leu Ala Lys Glu
515 520 525Asn Val Ala Ala Pro ★
530(2)序列6资料:
(i)序列特征:
(A)长度:2542个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(vi)来源:
(A)生物:稻
(ix)特征:
(A)名称/键:CDS
(B)位置:453..2282
(xi)序列描述:序列6:GAATTCAGTG TGAAGGAATA GATTCTCTTC AAAACAATTT AATCATTCAT CTGATCTGCT 60CAAAGCTCTG TGCATCTCCG GGTGCAACGG CCAGGATATT TATTGTGCAG TAAAAAAATG 120TCATATCCCC TAGCCACCCA AGAAACTGCT CCTTAAGTCC TTATAAGCAC ATATGGCATT 180GTAATATATA TGTTTGAGTT TTAGCGACAA TTTTTTTAAA AACTTTTGGT CCTTTTTATG 240AACGTTTTAA GTTTCACTGT CTTTTTTTTT CGAATTTTAA ATGTAGCTTC AAATTCTAAT 300CCCCAATCCA AATTGTAATA AACTTCAATT CTCCTAATTA ACATCTTAAT TCATTTATTT 360GAAAACCAGT TCAAATTCTT TTTAGGCTCA CCAAACCTTA AACAATTCAA TTCAGTGCAG 420AGATCTTCCA CAGCAACAGC TAGACAACCA CC ATG TCG GCT CTC ACC ACG TCC 473
Met Ser Ala Leu Thr Thr Ser
535 540CAG CTC GCC ACC TCG GCC ACC GGC TTC GGC ATC GCC GAC AGG TCG GCG 521Gln Leu Ala Thr Ser Ala Thr Gly Phe Gly Ile Ala Asp Arg Ser Ala
545 550 555CCG TCG TCG CTG CTC CGC CAC GGG TTC CAG GGC CTC AAG CCC CGC AGC 569Pro Ser Ser Leu Leu Arg His Gly Phe Gln Gly Leu Lys Pro Arg Ser
560 565 570CCC GCC GGC GGC GAC GCG ACG TCG CTC AGC GTG ACG ACC AGC GCG CGC 617Pro Ala Gly Gly Asp Ala Thr Ser Leu Ser Val Thr Thr Ser Ala Arg
575 580 585GCG ACG CCC AAG CAG CAG CGG TCG GTG CAG CGT GGC AGC CGG AGG TTC 665Ala Thr Pro Lys Gln Gln Arg Ser Val Gln Arg G1y Ser Arg Arg Phe590 595 600 605CCC TCC GTC GTC GTG TAC GCC ACC GGC GCC GGC ATG AAC GTC GTG TTC 713Pro Ser Val Val Val Tyr Ala Thr Gly Ala Gly Met Asn Val Val Phe
610 615 620GTC GGC GCC GAG ATG GCC CCC TGG AGC AAG ACC GGC GGC CTC GGT GAC 761Val Gly Ala Glu Met Ala Pro Trp Ser Lys Thr Gly Gly Leu Gly Asp
625 630 635GTC CTC GGT GGC CTC CCC CCT GCC ATG GCT GCG AAT GGC CAC AGG GTC 809Val Leu Gly Gly Leu Pro Pro Ala Met Ala Ala Asn Gly is Arg Val
640 645 650ATG GTG ATC TCT CCT CGG TAC GAC CAG TAC AAG GAC GCT TGG GAT ACC 857Met Val Ile Ser Pro Arg Tyr Asp Gln Tyr Lys Asp Ala Trp Asp Thr
655 660 665AGC GTT GTG GCT GAG ATC AAG GTT GCA GAC AGG TAC GAG AGG GTG AGG 905Ser Val Val Ala Glu Ile Lys Val Ala Asp Arg Tyr Glu Arg Val Arg670 675 680 685TTT TTC CAT TGC TAC AAG CGT GGA GTC GAC CGT GTG TTC ATC GAC CAT 953Phe Phe His Cys Tyr Lys Arg Gly Val Asp Arg Val Phe Ile Asp His
690 695 700CCG TCA TTC CTG GAG AAG GTT TGG GGA AAG ACC GGT GAG AAG ATC TAC 1001Pro Ser Phe Leu Glu Lys Val Trp Gly Lys Thr Gly Glu Lys Ile Tyr
705 710 715GGA CCT GAC ACT GGA GTT GAT TAC AAA GAC AAC CAG ATG CGT TTC AGC 1049Gly Pro Asp Thr Gly Val Asp Tyr Lys Asp Asn Gln Met Arg Phe Ser
720 725 730CTT CTT TGC CAG GCA GCA CTC GAG GCT CCT AGG ATC CTA AAC CTC AAC 1097Leu Leu Cys Gln Ala Ala Leu Glu Ala Pro Arg Ile Leu Asn Leu Asn
735 740 745AAC AAC CCA TAC TTC AAA GGA ACT TAT GGT GAG GAT GTT GTG TTC GTC 1145Asn Asn Pro Tyr Phe Lys Gly Thr Tyr Gly Glu Asp Val Val Phe Val750 755 760 765TGC AAC GAC TGG CAC ACT GGC CCA CTG GCG AGC TAC CTG AAG AAC AAC 1193Cys Asn Asp Trp His Thr Gly Pro Leu Ala Ser Tyr Leu Lys Asn Asn
770 775 780TAC CAG CCC AAT GGC ATC TAC AGG AAT GCA AAG GTT GCT TTC TGC ATC 1241Tyr Gln Pro Asn Gly Ile Tyr Arg Asn Ala Lys Val Ala Phe Cys Ile
785 790 795CAC AAC ATC TCC TAC CAG GGC CGT TTC GCT TTC GAG GAT TAC CCT GAG 1289His Asn Ile Ser Tyr Gln Gly Arg Phe Ala Phe Glu Asp Tyr Pro Glu
800 805 810CTG AAC CTC TCC GAG AGG TTC AGG TCA TCC TTC GAT TTC ATC GAC GGG 1337Leu Asn Leu Ser Glu Arg Phe Arg Ser Ser Phe Asp Phe Ile Asp Gly
815 820 825TAT GAC ACG CCG GTG GAG GGC AGG AAG ATC AAC TGG ATG AAG GCC GGA 1385Tyr Asp Thr Pro Val Glu Gly Arg Lys Ile Asn Trp Met Lys Ala Gly830 835 840 845ATC CTG GAA GCC GAC AGG GTG CTC ACC GTG AGC CCG TAC TAC GCC GAG 1433Ile Leu Glu Ala Asp Arg Val Leu Thr Val Ser Pro Tyr Tyr Ala Glu
850 855 860GAG CTC ATC TCC GGC ATC GCC AGG GGA TGC GAG CTC GAC AAC ATC ATG 1481Glu Leu Ile Ser Gly Ile Ala Arg Gly Cys Glu Leu Asp Asn Ile Met
865 870 875CGG CTC ACC GGC ATC ACC GGC ATC GTC AAC GGC ATG GAC GTC AGC GAG 1529Arg Leu Thr G1y Ile Thr Gly Ile Val Asn Gly Met Asp Val Ser Glu
880 885 890TGG GAT CCT AGC AAG GAC AAG TAC ATC ACC GCC AAG TAC GAC GCA ACC 1577Trp Asp Pro Ser Lys Asp Lys Tyr Ile Thr Ala Lys Tyr Asp Ala Thr
895 900 906ACG GCA ATC GAG GCG AAG GCG CTG AAC AAG GAG GCG TTG CAG GCG GAG 1625Thr Ala Ile Glu Ala Lys Ala Leu Asn Lys Glu Ala Leu Gln Ala Glu910 915 920 925GCG GGT CTT CCG GTC GAC AGG AAA ATC CCA CTG ATC GCG TTC ATC GGC 1673Ala Gly Leu Pro Val Asp Arg Lys Ile Pro Leu Ile Ala Phe Ile Gly
930 935 940AGG CTG GAG GAA CAG AAG GGC CCT GAC GTC ATG GCC GCC GCC ATC CCG 1721Arg Leu Glu Glu Gln Lys Gly Pro Asp Val Met Ala Ala Ala Ile Pro
945 950 955GAG CTC ATG CAG GAG GAC GTC CAG ATC GTT CTT CTG GGT ACT GGA AAG 1769Glu Leu Met Gln Glu Asp Val Gln Ile Val Leu Leu Gly Thr Gly Lys
960 965 970AAG AAG TTC GAG AAG CTG CTC AAG AGC ATG GAG GAG AAG TAT CCG GGC 1817Lys Lys Phe Glu Lys Leu Leu Lys Ser Met Glu Glu Lys Tyr Pro Gly
975 980 985AAG GTG AGG GCG GTG GTG AAG TTC AAC GCG CCG CTT GCT CAT CTC ATC 1865Lys Val Arg Ala Val Val Lys Phe Asn Ala Pro Leu Ala His Leu Ile990 995 1000 1005ATG GCC GGA GCC GAC GTG CTC GCC GTC CCC AGC CGC TTC GAG CCC TGT 1913Met Ala Gly Ala Asp Val Leu Ala Val Pro Ser Arg Phe Glu Pro Cys
1010 1015 1020GGA CTC ATC CAG CTG CAG GGG ATG AGA TAC GGA ACG CCC TGT GCT TGC 1961Gly Leu Ile Gln Leu Gln Gly Met Arg Tyr Gly Thr Pro Cys Ala Cys
1025 1030 1035GCG TCC ACC GGT GGG CTC GTG GAC ACG GTC ATC GAA GGC AAG ACT GGT 2009Ala Ser Thr Gly Gly Leu Val Asp Thr Val Ile Glu Gly Lys Thr Gly
1040 1045 1050TTC CAC ATG GGC CGT CTC AGC GTC GAC TGC AAG GTG GTG GAG CCA AGC 2057Phe His Met Gly Arg Leu Ser Val Asp Cys Lys Val Val Glu Pro Ser
1055 1060 1065GAC GTG AAG AAG GTG GCG GCC ACC CTG AAG CGC GCC ATC AAG GTC GTC 2105Asp Val Lys Lys Val Ala Ala Thr Leu Lys Arg Ala Ile Lys Val Val1070 1075 1080 1085GGC ACG CCG GCG TAC GAG GAG ATG GTC AGG AAC TGC ATG AAC CAG GAC 2153Gly Thr Pro Ala Tyr Glu Glu Met Val Arg Asn Cys Met Asn Gln Asp
1090 1095 1100CTC TCC TGG AAG GGG CCT GCG AAG AAC TGG GAG AAT GTG CTC CTG GGC 2201Leu Ser Trp Lys Gly Pro Ala Lys Asn Trp Glu Asn Val Leu Leu Gly
1105 1110 1115CTG GGC GTC GCC GGC AGC GCG CCG GGG ATC GAA GGC GAC GAG ATC GCG 2249Leu Gly Val Ala Gly Ser Ala Pro Gly Ile Glu Gly Asp Glu Ile Ala
1120 1125 1130CCG CTC GCC AAG GAG AAC GTG GCT GCT CCT TGA AGAGCCTGAG ATCTACATAT 2302Pro Leu Ala Lys Glu Asn Val Ala Ala Pro ★
1135 1140GGAGTGATTA ATTAATATAG CAGTATATGG ATGAGAGACG AATGAACCAG TGGTTTGTTT 2362GTTGTAGTGA ATTTGTAGCT ATAGCCAATT ATATAGGCTA ATAAGTTTGA TGTTGTACTC 2422TTCTGGGTGT GCTTAAGTAT CTTATCGGAC CCTGAATTTA TGTGTGTGGC TTATTGCCAA 2482TAATATTAAG TAATAAAGGG TTTATTATAT TATTATATAT GTTATATTAT ACTAAAAAAA 2542(2)序列7资料:
(i)序列特征:
(A)长度:6l0氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列7:Met Ser Ala Leu Thr Thr Ser Gln Leu Ala Thr Ser Ala Thr Gly Phe1 5 10 15Gly Ile Ala Asp Arg Ser Ala Pro Ser Ser Leu Leu Arg His Gly Phe
20 25 30Gln Gly Leu Lys Pro Arg Ser Pro Ala Gly Gly Asp Ala Thr Ser Leu
35 40 45Ser Val Thr Thr Ser Ala Arg Ala Thr Pro Lys Gln Gln Arg Ser Val
50 55 60Gln Arg Gly Ser Arg Arg Phe Pro Ser Val Val Val Tyr Ala Thr Gly65 70 75 80Ala Gly Met Asn Val Val Phe Val Gly Ala Glu Met Ala Pro Trp Ser
85 90 95Lys Thr Gly Gly Leu Gly Asp Val Leu Gly Gly Leu Pro Pro Ala Met
100 105 110Ala Ala Asn Gly His Arg Val Met Val Ile Ser Pro Arg Tyr Asp Gln
115 120 125Tyr Lys Asp Ala Trp Asp Thr Ser Val Val Ala Glu Ile Lys Val Ala
130 135 140Asp Arg Tyr Glu Arg Val Arg Phe Phe His Cys Tyr Lys Arg Gly Val145 150 155 160Asp Arg Val Phe Ile Asp His Pro Ser Phe Leu Glu Lys Val Trp Gly
165 170 175Lys Thr Gly Glu Lys Ile Tyr Gly Pro Asp Thr Gly Val Asp Tyr Lys
180 185 190Asp Asn Gln Met Arg Phe Ser Leu Leu Cys Gln Ala Ala Leu Glu Ala
195 200 205Pro Arg Ile Leu Asn Leu Asn Asn Asn Pro Tyr Phe Lys Gly Thr Tyr
210 215 220Gly Glu Asp Val Val Phe Val Cys Asn Asp Trp His Thr Gly Pro Leu225 230 235 240Ala Ser Tyr Leu Lys Asn Asn Tyr Gln Pro Asn Gly Ile Tyr Arg Asn
245 250 255Ala Lys Val Ala Phe Cys Ile His Asn Ile Ser Tyr Gln Gly Arg Phe
260 265 270Ala Phe Glu Asp Tyr Pro Glu Leu Asn Leu Ser Glu Arg Phe Arg Ser
275 280 285Ser Phe Asp Phe Ile Asp Gly Tyr Asp Thr Pro Val Glu Gly Arg Lys
290 295 300Ile Asn Trp Met Lys Ala Gly Ile Leu Glu Ala Asp Arg Val Leu Thr305 310 315 320Val Ser Pro Tyr Tyr Ala Glu Glu Leu Ile Ser Gly Ile Ala Arg Gly
325 330 335Cys Glu Leu Asp Asn Ile Met Arg Leu Thr Gly Ile Thr Gly Ile Val
340 345 350Asn Gly Met Asp Val Ser Glu Trp Asp Pro Ser Lys Asp Lys Tyr Ile
355 360 365Thr Ala Lys Tyr Asp Ala Thr Thr Ala Ile Glu Ala Lys Ala Leu Asn
370 375 380Lys Glu Ala Leu Gln Ala Glu Ala Gly Leu Pro Val Asp Arg Lys Ile385 390 395 400Pro Leu Ile Ala Phe Ile Gly Arg Leu Glu Glu Gln Lys Gly Pro Asp
405 410 415Val Met Ala Ala Ala Ile Pro Glu Leu Met Gln Glu Asp Val Gln Ile
420 425 430Val Leu Leu Gly Thr Gly Lys Lys Lys Phe Glu Lys Leu Leu Lys Ser
435 440 445Met Glu Glu Lys Tyr Pro Gly Lys Val Arg Ala Val Val Lys Phe Asn
450 455 460Ala Pro Leu Ala His Leu Ile Met Ala Gly Ala Asp Val Leu Ala Val465 470 475 480Pro Ser Arg Phe Glu Pro Cys Gly Leu Ile Gln Leu Gln Gly Met Arg
485 490 495Tyr Gly Thr Pro Cys Ala Cys Ala Ser Thr Gly Gly Leu Val Asp Thr
500 505 510Val Ile Glu Gly Lys Thr Gly Phe His Met Gly Arg Leu Ser Val Asp
515 520 525Cys Lys Val Val Glu Pro Ser Asp Val Lys Lys Val Ala Ala Thr Leu
530 535 540Lys Arg Ala Ile Lys Val Val Gly Thr Pro Ala Tyr Glu Glu Met Val545 550 555 560Arg Asn Cys Met Asn Gln Asp Leu Ser Trp Lys Gly Pro Ala Lys Asn
565 570 575Trp Glu Asn Val Leu Leu Gly Leu Gly Val Ala Gly Ser Ala Pro Gly
580 585 550Ile Glu Gly Asp Glu Ile Ala Pro Leu Ala Lys Glu Asn Val Ala Ala
595 600 605Pro ★
610(2)序列8资料:
(i)序列特征:
(A)长度:2007个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(vi)来源:
(A)生物:玉米
(ix)特征:
(A)名称/键:CDS
(B)位置:1..2007
(xi)序列描述:序列8:GCT GAG GCT GAG GCC GGG GGC AAG GAC GCG CCG CCG GAG AGG AGC GGC 48Ala Glu Ala Glu Ala Gly Gly Lys Asp Ala Pro Pro Glu Arg Ser Gly
615 620 625GAC GCC GCC AGG TTG CCC CGC GCT CGG GGC AAT GCG GTC TCC AAA CGG 96Asp Ala Ala Arg Leu Pro Arg Ala Arg Arg Asn Ala Val Ser Lys Arg
630 635 640AGG GAT CCT CTT CAG CCG GTC GGC CGG TAC GGC TCC GCG ACG GGA AAC 144Arg Asp Pro Leu Gln Pro Val Gly Arg Tyr Gly Ser Ala Thr Gly Asn
645 650 655ACG GCC AGG ACC GGC GCC GCG TCC TGC CAG AAC GCC GCA TTG GCG GAC 192Thr Ala Arg Thr Gly Ala Ala Ser Cys Gln Asn Ala Ala Leu Ala Asp
660 665 670GTT GAG ATC GTT GAG ATC AAG TCC ATC GTC GCC GCG CCG CCG ACG AGC 240Val Glu Ile Val Glu Ile Lys Ser Ile Val Ala Ala Pro Pro Thr Ser675 680 685 690ATA GTG AAG TTC CCA GGG CGC GGG CTA CAG GAT GAT CCT TCC CTC TGG 288Ile Val Lys Phe Pro Gly Arg Gly Leu Gln Asp Asp Pro Ser Leu Trp
695 700 705GAC ATA GCA CCG GAG ACT GTC CTC CCA GCC CCG AAG CCA CTG CAT GAA 336Asp Ile Ala Pro Glu Thr Val Leu Pro Ala Pro Lys Pro Leu His Glu
710 715 720TCG CCT GCG GTT GAC GGA GAT TCA AAT GGA ATT GCA CCT CCT ACA GTT 384Ser Pro Ala Val Asp Gly Asp Ser Asn Gly Ile Ala Pro Pro Thr Val
725 730 735GAG CCA TTA GTA CAG GAG GCC ACT TGG GAT TTC AAG AAA TAC ATC GGT 432Glu Pro Leu Val Gln Glu Ala Thr Trp Asp Phe Lys Lys Tyr Ile Gly
740 745 750TTT GAC GAG CCT GAC GAA GCG AAG GAT GAT TCC AGG GTT GGT GCA GAT 480Phe Asp Glu Pro Asp Glu Ala Lys Asp Asp Ser Arg Val Gly Ala Asp755 760 765 770GAT GCT GGT TCT TTT GAA CAT TAT GGG ACA ATG ATT CTG GGC CTT TGT 528Asp Ala Gly Ser Phe Glu His Tyr Gly Thr Met Ile Leu Gly Leu Cys
775 780 785GGG GAG AAT GTT ATG AAC GTG ATC GTG GTG GCT GCT GAA TGT TCT CCA 576Gly Glu Asn Val Met Asn Val Ile Val Val Ala Ala Glu Cys Ser Pro
790 795 800TGG TGC AAA ACA GGT GGT CTT GGA GAT GTT GTG GGA GCT TTA CCC AAG 624Trp Cys Lys Thr Gly Gly Leu Gly Asp Val Val Gly Ala Leu Pro Lys
805 810 815GCT TTA GCG AGA AGA GGA CAT CGT GTT ATG GTT GTG GTA CCA AGG TAT 672Ala Leu Ala Arg Arg Gly His Arg Val Met Val Val Val Pro Arg Tyr
820 825 830GGG GAC TAT GTG GAA GCC TTT GAT ATG GGA ATC CGG AAA TAC TAC AAA 720Gly Asp Tyr Val Glu Ala Phe Asp Met Gly Ile Arg Lys Tyr Tyr Lys835 840 845 850GCT GCA GGA CAG GAC CTA GAA GTG AAC TAT TTC CAT GCA TTT ATT GAT 768Ala Ala Gly Gln Asp Leu Glu Val Asn Tyr Phe His Ala Phe Ile Asp
855 860 865GGA GTC GAC TTT GTG TTC ATT GAT GCC TCT TTC CGG CAC CGT CAA GAT 816Gly Val Asp Phe Val Phe Ile Asp Ala Ser Phe Arg His Arg Gln Asp
870 875 880GAC ATA TAT GGG GGA AGT AGG CAG GAA ATC ATG AAG CGC ATG ATT TTG 864Asp Ile Tyr Gly Gly Ser Arg Gln Glu Ile Met Lys Arg Met Ile Leu
885 890 895TTT TGC AAG GTT GCT GTT GAG GTT CCT TGG CAC GTT CCA TGC GGT GGT 912Phe Cys Lys Val Ala Val Glu Val Pro Trp His Val Pro Cys Gly Gly
900 905 910GTG TGC TAC GGA GAT GGA AAT TTG GTG TTC ATT GCC ATG AAT TGG CAC 960Val Cys Tyr Gly Asp Gly Asn Leu Val Phe Ile Ala Met Asn Trp His915 920 925 930ACT GCA CTC CTG CCT GTT TAT CTG AAG GCA TAT TAC AGA GAC CAT GGG 1008Thr Ala Leu Leu Pro Val Tyr Leu Lys Ala Tyr Tyr Arg Asp His Gly
935 940 945TTA ATG CAG TAC ACT CGC TCC GTC CTC GTC ATA CAT AAC ATC GGC CAC 1056Leu Met Gln Tyr Thr Arg Ser Val Leu Val Ile His Asn Ile Gly His
950 955 960CAG GGC CGT GGT CCT GTA CAT GAA TTC CCG TAC ATG GAC TTG CTG AAC 1104Gln Gly Arg Gly Pro Val His Glu Phe Pro Tyr Met Asp Leu Leu Asn
965 970 975ACT AAC CTT CAA CAT TTC GAG CTG TAC GAT CCC GTC GGT GGC GAG CAC 1152Thr Asn Leu Gln His Phe Glu Leu Tyr Asp Pro Val Gly Gly Glu His
980 985 990GCC AAC ATC TTT GCC GCG TGT GTT CTG AAG ATG GCA GAC CGG GTG GTG 1200Ala Asn Ile Phe Ala Ala Cys Val Leu Lys Met Ala Asp Arg Val Val995 1000 1005 1010ACT GTC AGC CGC GGC TAC CTG TGG GAG CTG AAG ACA GTG GAA GGC GGC 1248Thr Val Ser Arg Gly Tyr Leu Trp Glu Leu Lys Thr Val Glu Gly Gly
1015 1020 1025TGG GGC CTC CAC GAC ATC ATC CGT TCT AAC GAC TGG AAG ATC AAT GGC 1296Trp Gly Leu His Asp Ile Ile Arg Ser Asn Asp Trp Lys Ile Asn Gly
1030 1035 1040ATT CGT GAA CGC ATC GAC CAC CAG GAG TGG AAC CCC AAG GTG GAC GTG 1344Ile Arg Glu Arg Ile Asp His Gln Glu Trp Asn Pro Lys Val Asp Val
1045 1050 1055CAC CTG CGG TCG GAC GGC TAC ACC AAC TAC TCC CTC GAG ACA CTC GAC 1392His Leu Arg Ser Asp Gly Tyr Thr Asn Tyr Ser Leu Glu Thr Leu Asp
1060 1065 1070GCT GGA AAG CGG CAG TGC AAG GCG GCC CTG CAG CGG GAC GTG GGC CTG 1440Ala Gly Lys Arg Gln Cys Lys Ala Ala Leu Gln Arg Asp Val Gly Leu1075 1080 1085 1090GAA GTG CGC GAC GAC GTG CCG CTG CTC GGC TTC ATC GGG CGT CTG GAT 1488Glu Val Arg Asp Asp Val Pro Leu Leu Gly Phe Ile Gly Arg Leu Asp
1095 1100 1105GGA CAG AAG GGC GTG GAC ATC ATC GGG GAC GCG ATG CCG TGG ATC GCG 1536Gly Gln Lys Gly Val Asp Ile Ile Gly Asp Ala Met Pro Trp Ile Ala
1110 1115 1120GGG CAG GAC GTG CAG CTG GTG ATG CTG GGC ACC GGC CCA CCT GAC CTG 1584Gly Gln Asp Val Gln Leu Val Met Leu Gly Thr Gly Pro Pro Asp Leu
1125 1130 1135GAA CGA ATG CTG CAG CAC TTG GAG CGG GAG CAT CCC AAC AAG GTG CGC 1632Glu Arg Met Leu Gln His Leu Glu Arg Glu His Pro Asn Lys Val Arg
1140 1145 1150GGG TGG GTC GGG TTC TCG GTC CTA ATG GTG CAT CGC ATC ACG CCG GGC 1680Gly Trp Val Gly Phe Ser Val Leu Met Val His Arg Ile Thr Pro Gly1155 1160 1165 1170GCC AGC GTG CTG GTG ATG CCC TCC CGC TTC GCC GGC GGG CTG AAC CAG 1728Ala Ser Val Leu Val Met Pro Ser Arg Phe Ala Gly Gly Leu Asn Gln
1175 1180 1185CTC TAC GCG ATG GCA TAC GGC ACC GTC CCT GTG GTG CAC GCC GTG GGC 1776Leu Tyr Ala Met Ala Tyr Gly Thr Val Pro Val Val His Ala Val Gly
1190 1195 1200GGG CTC AGG GAC ACC GTG GCG CCG TTC GAC CCG TTC GGC GAC GCC GGG 1824Gly Leu Arg Asp Thr Val Ala Pro Phe Asp Pro Phe Gly Asp Ala Gly
1205 1210 1215CTC GGG TGG ACT TTT GAC CGC GCC GAG GCC AAC AAG CTG ATC GAG GTG 1872Leu Gly Trp Thr Phe Asp Arg Ala Glu Ala Asn Lys Leu Ile Glu Val
1220 1225 1230CTC AGC CAC TGC CTC GAC ACG TAC CGA AAC TAC GAG GAG AGC TGG AAG 1920Leu Ser His Cys Leu Asp Thr Tyr Arg Asn Tyr Glu Glu Ser Trp Lys1235 1240 1245 1250AGT CTC CAG GCG CGC GGC ATG TCG CAG AAC CTC AGC TGG GAC CAC GCG 1968Ser Leu Gln Ala Arg Gly Met Ser Gln Asn Leu Ser Trp Asp His Ala
1255 1260 1265GCT GAG CTC TAC GAG GAC GTC CTT GTC AAG TAC CAG TGG 2007Ala Glu Leu Tyr Glu Asp Val Leu Val Lys Tyr Gln Trp
1270 1275(2)序列9资料:
(i)序列特征:
(A)长度:669氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列9:Ala Glu Ala Glu Ala Gly Gly Lys Asp Ala Pro ProGlu Arg Ser Gly 1 5 10 15Asp Ala Ala Arg Leu Pro Arg Ala Arg Arg Asn Ala Val Ser Lys Arg
20 25 30Arg Asp Pro Leu Gln Pro Val Gly Arg Tyr Gly Ser Ala Thr Gly Asn
35 40 45Thr Ala Arg Thr Gly Ala Ala Ser Cys Gln Asn Ala Ala Leu Ala Asp
50 55 60Val Glu Ile Val Glu Ile Lys Ser Ile Val Ala Ala Pro Pro Thr Ser65 70 75 80Ile Val Lys Phe Pro Gly Arg Gly Leu Gln Asp Asp Pro Ser Leu Trp
85 90 95Asp Ile Ala Pro Glu Thr Val Leu Pro Ala Pro Lys Pro Leu His Glu
100 105 110Ser Pro Ala Val Asp Gly Asp Ser Asn Gly Ile Ala Pro Pro Thr Val
115 120 125Glu Pro Leu Val Gln Glu Ala Thr Trp Asp Phe Lys Lys Tyr Ile Gly
130 135 140Phe Asp Glu Pro Asp Glu Ala Lys Asp Asp Ser Arg Val Gly Ala Asp145 150 155 160Asp Ala Gly ser Phe Glu His Tyr Gly Thr Met Ile Leu Gly Leu Cys
165 170 175Gly Glu Asn Val Met Asn Val Ile Val Val Ala Ala Glu Cys Ser Pro
180 185 190Trp Cys Lys Thr Gly Gly Leu Gly Asp Val Val Gly Ala Leu Pro Lys
195 200 205Ala Leu Ala Arg Arg Gly His Arg Val Met Val Val Val Pro Arg Tyr
210 215 220Gly Asp Tyr Val Glu Ala Phe Asp Met Gly Ile Arg Lys Tyr Tyr Lys225 230 235 240Ala Ala Gly Gln Asp Leu Glu Val Asn Tyr Phe His Ala Phe Ile Asp
245 250 255Gly Val Asp Phe Val Phe Ile Asp Ala Ser Phe Arg His Arg Gln Asp
260 265 270Asp Ile Tyr Gly Gly Ser Arg Gln Glu Ile Met Lys Arg Met Ile Leu
275 280 285Phe Cys Lys Val Ala Val Glu Val Pro Trp His Val Pro Cys Gly Gly
290 295 300Val Cys Tyr Gly Asp Gly Asn Leu Val Phe Ile Ala Met Asn Trp His305 310 315 320Thr Ala Leu Leu Pro Val Tyr Leu Lys Ala Tyr Tyr Arg Asp His Gly
325 330 335Leu Met Gln Tyr Thr Arg Ser Val Leu Val Ile His Asn Ile Gly His
340 345 350Gln Gly Arg Gly Pro Val His Glu Phe Pro Tyr Met Asp Leu Leu Asn
355 360 365Thr Asn Leu Gln His Phe Glu Leu Tyr Asp Pro Val Gly Gly Glu His
370 375 380Ala Asn Ile Phe Ala Ala Cys Val Leu Lys Met Ala Asp Arg Val Val385 390 395 400Thr Val Ser Arg Gly Tyr Leu Trp Glu Leu Lys Thr Val Glu Gly Gly
405 410 415Trp Gly Leu His Asp Ile Ile Arg Ser Asn Asp Trp Lys Ile Asn Gly
420 425 430Ile Arg Glu Arg Ile Asp His Gln Glu Trp Asn Pro Lys Val Asp Val
435 440 445His Leu Arg Ser Asp Gly Tyr Thr Asn Tyr Ser Leu Glu Thr Leu Asp
450 455 460Ala Gly Lys Arg Gln Cys Lys Ala Ala Leu Gln Arg Asp Val Gly Leu465 470 475 480Glu Val Arg Asp Asp Val Pro Leu Leu Gly Phe Ile Gly Arg Leu Asp
485 490 495Gly Gln Lys Gly Val Asp Ile Ile Gly Asp Ala Met Pro Trp Ile Ala
500 505 510Gly Gln Asp Val Gln Leu Val Met Leu Gly Thr Gly Pro Pro Asp Leu
515 520 525Glu Arg Met Leu Gln His Leu Glu Arg Glu His Pro Asn Lys Val Arg
530 535 540Gly Trp Val Gly Phe Ser Val Leu Met Val His Arg Ile Thr Pro Gly545 550 555 560Ala Ser Val Leu Val Met Pro Ser Arg Phe Ala Gly Gly Leu Asn Gln
565 570 575Leu Tyr Ala Met Ala Tyr Gly Thr Val Pro Val Val His Ala Val Gly
580 585 590Gly Leu Arg Asp Thr Val Ala Pro Phe Asp Pro Phe Gly Asp Ala Gly
595 600 605Leu Gly Trp Thr Phe Asp Arg Ala Glu Ala Asn Lys Leu Ile Glu Val
610 615 620Leu Ser His Cys Leu Asp Thr Tyr Arg Asn Tyr Glu Glu Ser Trp Lys625 630 635 640Ser Leu Gln Ala Arg Gly Met Ser Gln Asn Leu Ser Trp Asp His Ala
645 650 655Ala Glu Leu Tyr Glu Asp Val Leu Val Lys Tyr Gln Trp
660 665(2)序列10资料:
(i)序列特征:
(A)长度:2097个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(vi)来源:
(A)生物:玉米
(ix)特征:
(A)名称/键:CDS
(B)位置:1..2097
(xi)序列描述:序列10:ATG CCG GGG GCA ATC TCT TCC TCG TCG TCG GCT TTT CTC CTC CCC GTC 48Met Pro Gly Ala Ile Ser Ser Ser Ser Ser Ala Phe Leu Leu Pro Val670 675 680 685GCG TCC TCC TCG CCG CGG CGC AGG CGG GGC AGT GTG GGT GCT GCT CTG 96Ala Ser Ser Ser Pro Arg Arg Arg Arg Gly Ser Val Gly Ala Ala Leu
690 695 700CGC TCG TAC GGC TAC AGC GGC GCG GAG CTG CGG TTG CAT TGG GCG CGG 144Arg Ser Tyr Gly Tyr Ser Gly Ala Glu Leu Arg Leu His Trp Ala Arg
705 710 715CGG GGC CCG CCT CAG GAT GGA GCG GCG TCG GTA CGC GCC GCA GCG GCA 192Arg Gly Pro Pro Gln Asp Gly Ala Ala Ser Val Arg Ala Ala Ala Ala
720 725 730CCG GCC GGG GGC GAA AGC GAG GAG GCA GCG AAG AGC TCC TCC TCG TCC 240Pro Ala Gly Gly Glu Ser Glu Glu Ala Ala Lys Ser Ser Ser Ser Ser
735 740 745CAG GCG GGC GCT GTT CAG GGC AGC ACG GCC AAG GCT GTG GAT TCT GCT 288Gln Ala Gly Ala Val Gln Gly Ser Thr Ala Lys Ala Val Asp Ser Ala750 755 760 765TCA CCT CCC AAT CCT TTG ACA TCT GCT CCG AAG CAA AGT CAG AGC GCT 336Ser Pro Pro Asn Pro Leu Thr Ser Ala Pro Lys Gln Ser Gln Ser Ala
770 775 780GCA ATG CAA AAC GGA ACG AGT GGG GGC AGC AGC GCG AGC ACC GCC GCG 384Ala Met Gln Asn Gly Thr Ser Gly Gly Ser Ser Ala Ser Thr Ala Ala
785 790 795CCG GTG TCC GGA CCC AAA GCT GAT CAT CCA TCA GCT CCT GTC ACC AAG 432Pro Val Ser Gly Pro Lys Ala Asp His Pro Ser Ala Pro Val Thr Lys
800 805 810AGA GAA ATC GAT GCC AGT GCG GTG AAG CCA GAG CCC GCA GGT GAT GAT 480Arg Glu Ile Asp Ala Ser Ala Val Lys Pro Glu Pro Ala Gly Asp Asp
815 820 825GCT AGA CCG GTG GAA AGC ATA GGC ATC GCT GAA CCG GTG GAT GCT AAG 528Ala Arg Pro Val Glu Ser Ile Gly Ile Ala Glu Pro Val Asp Ala Lys830 835 840 845GCT GAT GCA GCT CCG GCT ACA GAT GCG GCG GCG AGT GCT CCT TAT GAC 576Ala Asp Ala Ala Pro Ala Thr Asp Ala Ala Ala Ser Ala Pro Tyr Asp
850 855 860AGG GAG GAT AAT GAA CCT GGC CCT TTG GCT GGG CCT AAT GTG ATG AAC 624Arg Glu Asp Asn Glu Pro Gly Pro Leu Ala Gly Pro Asn Val Met Asn
865 870 875GTC GTC GTG GTG GCT TCT GAA TGT GCT CCT TTC TGC AAG ACA GGT GGC 672Val Val Val Val Ala Ser Glu Cys Ala Pro Phe Cys Lys Thr Gly Gly
880 885 890CTT GGA GAT GTC GTG GGT GCT TTG CCT AAG GCT CTG GCG AGG AGA GGA 720Leu Gly Asp Val Val Gly Ala Leu Pro Lys Ala Leu Ala Arg Arg Gly
895 900 905CAC CGT GTT ATG GTC GTG ATA CCA AGA TAT GGA GAG TAT GCC GAA GCC 768His Arg Val Met Val Val Ile Pro Arg Tyr Gly Glu Tyr Ala Glu Ala910 915 920 925CGG GAT TTA GGT GTA AGG AGA CGT TAC AAG GTA GCT GGA CAG GAT TCA 816Arg Asp Leu Gly Val Arg Arg Arg Tyr Lys Val Ala Gly Gln Asp Ser
930 935 940GAA GTT ACT TAT TTT CAC TCT TAC ATT GAT GGA GTT GAT TTT GTA TTC 864Glu Val Thr Tyr Phe His Ser Tyr Ile Asp Gly Val Asp Phe Val Phe
945 950 955GTA GAA GCC CCT CCC TTC CGG CAC CGG CAC AAT AAT ATT TAT GGG GGA 912Val Glu Ala Pro Pro Phe Arg His Arg His Asn Asn Ile Tyr Gly Gly
960 965 970GAA AGA TTG GAT ATT TTG AAG CGC ATG ATT TTG TTC TGC AAG GCC GCT 960Glu Arg Leu Asp Ile Leu Lys Arg Met Ile Leu Phe Cys Lys Ala Ala
975 980 985GTT GAG GTT CCA TGG TAT GCT CCA TGT GGC GGT ACT GTC TAT GGT GAT 1008Val Glu Val Pro Trp Tyr Ala Pro Cys Gly Gly Thr Val Tyr Gly Asp990 995 1000 1005GGC AAC TTA GTT TTC ATT GCT AAT GAT TGG CAT ACC GCA CTT CTG CCT 1056Gly Asn Leu Val Phe Ile Ala Asn Asp Trp His Thr Ala Leu Leu Pro
1010 1015 1020GTC TAT CTA AAG GCC TAT TAC CGG GAC AAT GGT TTG ATG CAG TAT GCT 1104Val Tyr Leu Lys Ala Tyr Tyr Arg Asp Asn Gly Leu Met Gln Tyr Ala
1025 1030 1035CGC TCT GTG CTT GTG ATA CAC AAC ATT GCT CAT CAG GGT CGT GGC CCT 1152Arg Ser Val Leu Val Ile His Asn Ile Ala His Gln Gly Arg Gly Pro
1040 1045 1050GTA GAC GAC TTC GTC AAT TTT GAC TTG CCT GAA CAC TAC ATC GAC CAC 1200Val Asp Asp Phe Val Asn Phe Asp Leu Pro Glu His Tyr Ile Asp His
1055 1060 1065TTC AAA CTG TAT GAC AAC ATT GGT GGG GAT CAC AGC AAC GTT TTT GCT 1248Phe Lys Leu Tyr Asp Asn Ile Gly Gly Asp His Ser Asn Val Phe Ala1070 1075 1080 1085GCG GGG CTG AAG ACG GCA GAC CGG GTG GTG ACC GTT AGC AAT GGC TAC 1296Ala Gly Leu Lys Thr Ala Asp Arg Val Val Thr Val Ser Asn Gly Tyr
1090 1095 1100ATG TGG GAG CTG AAG ACT TCG GAA GGC GGG TGG GGC CTC CAC GAC ATC 1344Met Trp Glu Leu Lys Thr Ser Glu Gly Gly Trp Gly Leu His Asp Ile
1105 1110 1115ATA AAC CAG AAC GAC TGG AAG CTG CAG GGC ATC GTG AAC GGC ATC GAC 1392Ile Asn Gln Asn Asp Trp Lys Leu Gln Gly Ile Val Asn Gly Ile Asp
1120 1125 1130ATG AGC GAG TGG AAC CCC GCT GTG GAC GTG CAC CTC CAC TCC GAC GAC 1440Met Ser Glu Trp Asn Pro Ala Val Asp Val His Leu His Ser Asp Asp
1135 1140 1145TAC ACC AAC TAC ACG TTC GAG ACG CTG GAC ACC GGC AAG CGG CAG TGC 1488Tyr Thr Asn Tyr Thr Phe Glu Thr Leu Asp Thr Gly Lys Arg Gln Cys1150 1155 1160 1165AAG GCC GCC CTG CAG CGG CAG CTG GGC CTG CAG GTC CGC GAC GAC GTG 1536Lys Ala Ala Leu Gln Arg Gln Leu Gly Leu Gln Val Arg Asp Asp Val
1170 1175 1180CCA CTG ATC GGG TTC ATC GGG CGG CTG GAC CAC CAG AAG GGC GTG GAC 1584Pro Leu Ile Gly Phe Ile Gly Arg Leu Asp His Gln Lys Gly Val Asp
1185 1190 1195ATC ATC GCC GAC GCG ATC CAC TGG ATC GCG GGG CAG GAC GTG CAG CTC 1632Ile Ile Ala Asp Ala Ile His Trp Ile Ala Gly Gln Asp Val Gln Leu
1200 1205 1210GTG ATG CTG GGC ACC GGG CGG GCC GAC CTG GAG GAC ATG CTG CGG CGG 1680Val Met Leu Gly Thr Gly Arg Ala Asp Leu Glu Asp Met Leu Arg Arg
1215 1220 1225TTC GAG TCG GAG CAC AGC GAC AAG GTG CGC GCG TGG GTG GGG TTC TCG 1728Phe Glu Ser Glu His Ser Asp Lys Val Arg Ala Trp Val Gly Phe Ser1230 1235 1240 1245GTG CCC CTG GCG CAC CGC ATC ACG GCG GGC GCG GAC ATC CTG CTG ATG 1776Val Pro Leu Ala His Arg Ile Thr Ala Gly Ala Asp Ile Leu Leu Met
1250 1255 1260CCG TCG CGG TTC GAG CCG TGC GGG CTG AAC CAG CTC TAC GCC ATG GCG 1824Pro Ser Arg Phe Glu Pro Cys Gly Leu Asn Gln Leu Tyr Ala Met Ala
1265 1270 1275TAC GGG ACC GTG CCC GTG GTG CAC GCC GTG GGG GGG CTC CGG GAC ACG 1872Tyr Gly Thr Val Pro Val Val His Ala Val Gly Gly Leu Arg Asp Thr
1280 1285 1290GTG GCG CCG TTC GAC CCG TTC AAC GAC ACC GGG CTC GGG TGG ACG TTC 1920Val Ala Pro Phe Asp Pro Phe Asn Asp Thr Gly Leu Gly Trp Thr Phe
1295 1300 1305GAC CGC GCG GAG GCG AAC CGG ATG ATC GAC GCG CTC TCG CAC TGC CTC 1968Asp Arg Ala Glu Ala Asn Arg Met Ile Asp Ala Leu Ser His Cys Leu1310 1315 1320 1325ACC ACG TAC CGG AAC TAC AAG GAG AGC TGG CGC GCC TGC AGG GCG CGC 2016Thr Thr Tyr Arg Asn Tyr Lys Glu Ser Trp Arg Ala Cys Arg Ala Arg
1330 1335 1340GGC ATG GCC GAG GAC CTC AGC TGG GAC CAC GCC GCC GTG CTG TAT GAG 2064Gly Met Ala Glu Asp Leu Ser Trp Asp His Ala Ala Val Leu Tyr Glu
1345 1350 1355GAC GTG CTC GTC AAG GCG AAG TAC CAG TGG TGA 2097Asp Val Leu Val Lys Ala Lys Tyr Gln Trp ★
1360 1365(2)序列11资料:
(i)序列特征:
(A)长度:699氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列11:Met Pro Gly Ala Ile Ser Ser Ser Ser Ser Ala Phe Leu Leu Pro Val1 5 10 15Ala Ser Ser Ser Pro Arg Arg Arg Arg Gly Ser Val Gly Ala Ala Leu
20 25 30Arg Ser Tyr Gly Tyr Ser Gly Ala Glu Leu Arg Leu His Trp Ala Arg
35 40 45Arg Gly Pro Pro Gln Asp Gly Ala Ala Ser Val Arg Ala Ala Ala Ala
50 55 60Pro Ala Gly Gly Glu Ser Glu Glu Ala Ala Lys Ser Ser Ser Ser Ser65 70 75 80Gln Ala Gly Ala Val Gln Gly Ser Thr Ala Lys Ala Val Asp Ser Ala
85 90 95Ser Pro Pro Asn Pro Leu Thr Ser Ala Pro Lys Gln Ser Gln Ser Ala
100 105 110Ala Met Gln Asn Gly Thr Ser Gly Gly Ser Ser Ala Ser Thr Ala Ala
115 120 125Pro Val Ser Gly Pro Lys Ala Asp His Pro Ser Ala Pro Val Thr Lye
130 135 140Arg Glu Ile Asp Ala Ser Ala Val Lys Pro Glu Pro Ala Gly Asp Asp145 150 155 160Ala Arg Pro Val Glu Ser Ile Gly Ile Ala Glu Pro Val Asp Ala Lys
165 170 175Ala Asp Ala Ala Pro Ala Thr Asp Ala Ala Ala Ser Ala Pro Tyr Asp
180 185 190Arg Glu Asp Asn Glu Pro Gly Pro Leu Ala Gly Pro Asn Val Met Asn
195 200 205Val Val Val Val Ala Ser Glu Cys Ala Pro Phe Cys Lys Thr Gly Gly
210 215 220Leu Gly Asp Val Val Gly Ala Leu Pro Lys Ala Leu Ala Arg Arg Gly225 230 235 240His Arg Val Met Val Val Ile Pro Arg Tyr Gly Glu Tyr Ala Glu Ala
245 250 255Arg Asp Leu Gly Val Arg Arg Arg Tyr Lys Val Ala Gly Gln Asp Ser
260 265 270Glu Val Thr Tyr Phe His Ser Tyr Ile Asp Gly Val Asp Phe Val Phe
275 280 285Val Glu Ala Pro Pro Phe Arg His Arg His Asn Asn Ile Tyr Gly Gly
290 295 300Glu Arg Leu Asp Ile Leu Lys Arg Met Ile Leu Phe Cys Lys Ala Ala305 310 315 320Val Glu Val Pro Trp Tyr Ala Pro Cys Gly Gly Thr Val Tyr Gly Asp
325 330 335Gly Asn Leu Val Phe Ile Ala Asn Asp Trp His Thr Ala Leu Leu Pro
340 345 350Val Tyr Leu Lys Ala Tyr Tyr Arg Asp Asn Gly Leu Met Gln Tyr Ala
355 360 365Arg Ser Val Leu Val Ile His Asn Ile Ala His Gln Gly Arg Gly Pro
370 375 380Val Asp Asp Phe Val Asn Phe Asp Leu Pro Glu His Tyr Ile Asp His385 390 395 400Phe Lys Leu Tyr Asp Asn Ile Gly Gly Asp His Ser Asn Val Phe Ala
405 410 415Ala Gly Leu Lys Thr Ala Asp Arg Val Val Thr Val Ser Asn Gly Tyr
420 425 430Met Trp Glu Leu Lys Thr Ser Glu Gly Gly Trp Gly Leu His Asp Ile
435 440 445Ile Asn Gln Asn Asp Trp Lys Leu Gln Gly Ile Val Asn Gly Ile Asp
450 455 460Met Ser Glu Trp Asn Pro Ala Val Asp Val His Leu His Ser Asp Asp465 470 475 480Tyr Thr Asn Tyr Thr Phe Glu Thr Leu Asp Thr Gly Lys Arg Gln Cys
485 490 495Lys Ala Ala Leu Gln Arg Gln Leu Gly Leu Gln Val Arg Asp Asp Val
500 505 510Pro Leu Ile Gly Phe Ile Gly Arg Leu Asp His Gln Lys Gly Val Asp
515 520 525Ile Ile Ala Asp Ala Ile His Trp Ile Ala Gly Gln Asp Val Gln Leu
530 535 540Val Met Leu Gly Thr Gly Arg Ala Asp Leu Glu Asp Met Leu Arg Arg545 550 555 560Phe Glu Ser Glu His Ser Asp Lys Val Arg Ala Trp Val Gly Phe Ser
565 570 575Val Pro Leu Ala His Arg Ile Thr Ala Gly Ala Asp Ile Leu Leu Met
580 585 590Pro Ser Arg Phe Glu Pro Cys Gly Leu Asn Gln Leu Tyr Ala Met Ala
595 600 605Tyr Gly Thr Val Pro Val Val His Ala Val Gly Gly Leu Arg Asp Thr
610 615 620Val Ala Pro Phe Asp Pro Phe Asn Asp Thr Gly Leu Gly Trp Thr Phe625 630 635 640Asp Arg Ala Glu Ala Asn Arg Met Ile Asp Ala Leu Ser His Cys Leu
645 650 655Thr Thr Tyr Arg Asn Tyr Lys Glu Ser Trp Arg Ala Cys Arg Ala Arg
660 665 670Gly Met Ala Glu Asp Leu Ser Trp Asp His Ala Ala Val Leu Tyr Glu
675 680 685Asp Val Leu Val Lys Ala Lys Tyr Gln Trp ★
690 695(2)序列12资料:
(i)序列特征:
(A)长度:1752个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(vi)来源:
(A)生物:玉米
(ix)特征:
(A)名称/键:CDS
(B)位置:1..1752
(xi)序列描述:序列12:TGC GTC GCG GAG CTG AGC AGG GAG GGG CCC GCG CCG CGC CCG CTG CCA 48Cys Val Ala Glu Leu Ser Arg Glu Gly Pro Ala Pro Arg Pro Leu Pro700 705 710 715CCC GCG CTG CTG GCG CCC CCG CTC GTG CCC GGC TTC CTC GCG CCG CCG 96Pro Ala Leu Leu Ala Pro Pro Leu Val Pro Gly Phe Leu Ala Pro Pro
720 725 730GCC GAG CCC ACG GGT GAG CCG GCA TCG ACG CCG CCG CCC GTG CCC GAC 144Ala Glu Pro Thr Gly Glu Pro Ala Ser Thr Pro Pro Pro Val Pro Asp
735 740 745GCC GGC CTG GGG GAC CTC GGT CTC GAA CCT GAA GGG ATT GCT GAA GGT 192Ala Gly Leu Gly Asp Leu Gly Leu Glu Pro Glu Gly Ile Ala Glu Gly
750 755 760TCC ATC GAT AAC ACA GTA GTT GTG GCA AGT GAG CAA GAT TCT GAG ATT 240Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln Asp Ser Glu Ile
765 770 775GTG GTT GGA AAG GAG CAA GCT CGA GCT AAA GTA ACA CAA AGC ATT GTC 288Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr Gln Ser Ile Val780 785 790 795TTT GTA ACC GGC GAA GCT TCT CCT TAT GCA AAG TCT GGG GGT CTA GGA 336Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser Gly Gly Leu Gly
800 805 810GAT GTT TGT GGT TCA TTG CCA GTT GCT CTT GCT GCT CGT GGT CAC CGT 384Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala Arg Gly His Arg
815 820 825GTG ATG GTT GTA ATG CCC AGA TAT TTA AAT GGT ACC TCC GAT AAG AAT 432Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr Ser Asp Lys Asn
830 835 840TAT GCA AAT GCA TTT TAC ACA GAA AAA CAC ATT CGG ATT CCA TGC TTT 480Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg Ile Pro Cys Phe
845 850 855GGC GGT GAA CAT GAA GTT ACC TTC TTC CAT GAG TAT AGA GAT TCA GTT 528Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr Arg Asp Ser Val860 865 870 875GAC TGG GTG TTT GTT GAT CAT CCC TCA TAT CAC AGA CCT GGA AAT TTA 576Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg Pro Gly Asn Leu
880 885 890TAT GGA GAT AAG TTT GGT GCT TTT GGT GAT AAT CAG TTC AGA TAC ACA 624Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln Phe Arg Tyr Thr
895 900 905CTC CTT TGC TAT GCT GCA TGT GAG GCT CCT TTG ATC CTT GAA TTG GGA 672Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile Leu Glu Leu Gly
910 915 920GGA TAT ATT TAT GGA CAG AAT TGC ATG TTT GTT GTC AAT GAT TGG CAT 720Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val Asn Asp Trp His
925 930 935GCC AGT CTA GTG CCA GTC CTT CTT GCT GCA AAA TAT AGA CCA TAT GGT 768Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr Arg Pro Tyr Gly940 945 950 955GTT TAT AAA GAC TCC CGC AGC ATT CTT GTA ATA CAT AAT TTA GCA CAT 816Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile His Asn Leu Ala His
960 965 970CAG GGT GTA GAG CCT GCA AGC ACA TAT CCT GAC CTT GGG TTG CCA CCT 864Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu Gly Leu Pro Pro
975 980 985GAA TGG TAT GGA GCT CTG GAG TGG GTA TTC CCT GAA TGG GCG AGG AGG 912Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu Trp Ala Arg Arg
990 995 1000CAT GCC CTT GAC AAG GGT GAG GCA GTT AAT TTT TTG AAA GGT GCA GTT 960His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu Lys Gly Ala Val
1005 1010 1015GTG ACA GCA GAT CGA ATC GTG ACT GTC AGT AAG GGT TAT TCG TGG GAG 1008Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly Tyr Ser Trp Glu1020 1025 1030 1035GTC ACA ACT GCT GAA GGT GGA CAG GGC CTC AAT GAG CTC TTA AGC TCC 1056Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu Leu Leu Ser Ser
1040 1045 1050AGA AAG AGT GTA TTA AAC GGA ATT GTA AAT GGA ATT GAC ATT AAT GAT 1104Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile Asp Ile Asn Asp
1055 1060 1065TGG AAC CCT GCC ACA GAC AAA TGT ATC CCC TGT CAT TAT TCT GTT GAT 1152Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His Tyr Ser Val Asp
1070 1075 1080GAC CTC TCT GGA AAG GCC AAA TGT AAA GGT GCA TTG CAG AAG GAG CTG 1200Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu Gln Lys Glu Leu
1085 1090 1095GGT TTA CCT ATA AGG CCT GAT GTT CCT CTG ATT GGC TTT ATT GGA AGG 1248Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly Phe Ile Gly Arg1100 1105 1110 1115TTG GAT TAT CAG AAA GGC ATT GAT CTC ATT CAA CTT ATC ATA CCA GAT 1296Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu Ile Ile Pro Asp
1120 1125 1130CTC ATG CGG GAA GAT GTT CAA TTT GTC ATG CTT GGA TCT GGT GAC CCA 1344Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly Ser Gly Asp Pro
1135 1140 1145GAG CTT GAA GAT TGG ATG AGA TCT ACA GAG TCG ATC TTC AAG GAT AAA 1392Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile Phe Lys Asp Lys
1150 1155 1160TTT CGT GGA TGG GTT GGA TTT AGT GTT CCA GTT TCC CAC CGA ATA ACT 1440Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser His Arg Ile Thr
1165 1170 1175GCC GGC TGC GAT ATA TTG TTA ATG CCA TCC AGA TTC GAA CCT TGT GGT 1488Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe Glu Pro Cys Gly1180 1185 1190 1195CTC AAT CAG CTA TAT GCT ATG CAG TAT GGC ACA GTT CCT GTT GTC CAT 1536Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val Pro Val Val His
1200 1205 1210GCA ACT GGG GGC CTT AGA GAT ACC GTG GAG AAC TTC AAC CCT TTC GGT 1584Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe Asn Pro Phe Gly
1215 1220 1225GAG AAT GGA GAG CAG GGT ACA GGG TGG GCA TTC GCA CCC CTA ACC ACA 1632Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala Pro Leu Thr Thr
1230 1235 1240GAA AAC ATG TTT GTG GAC ATT GCG AAC TGC AAT ATC TAC ATA CAG GGA 1680Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile Tyr Ile Gln Gly
1245 1250 1255ACA CAA GTC CTC CTG GGA AGG GCT AAT GAA GCG AGG CAT GTC AAA AGA 1728Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg His Val Lys ArgT 1260 1265 1270 1275CTT CAC GTG GGA CCA TGC CGC TGA 1752Leu His Val Gly Pro Cys Arg ★
1280(2)序列13资料:
(i)序列特征:
(A)长度:584氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列13:Cys Val Ala Glu Leu Ser Arg Glu Gly Pro Ala Pro Arg Pro Leu Pro1 5 10 15Pro Ala Leu Leu Ala Pro Pro Leu Val Pro Gly Phe Leu Ala Pro Pro
20 25 30Ala Glu Pro Thr Gly Glu Pro Ala Ser Thr Pro Pro Pro Val Pro Asp
35 40 45Ala Gly Leu Gly Asp Leu Gly Leu Glu Pro Glu Gly Ile Ala Glu Gly
50 55 60Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln Asp Ser Glu Ile65 70 75 80Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr Gln Ser Ile Val
85 90 95Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser Gly Gly Leu Gly
100 105 110Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala Arg Gly His Arg
115 120 125Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr Ser Asp Lys Asn
130 135 140Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg Ile Pro Cys Phe145 150 155 160Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr Arg Asp Ser Val
165 170 175Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg Pro Gly Asn Leu
180 185 190Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln Phe Arg Tyr Thr
195 200 205Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile Leu Glu Leu Gly
210 215 220Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val Asn Asp Trp His225 230 235 240Aia Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr Arg Pro Tyr Gly
245 250 255Val Tyr Lys Asp Ser Arg Ser Ile Leu Val IleHis Asn Leu Ala His
260 265 270Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu Gly Leu Pro Pro
275 280 285Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu Trp Ala Arg Arg
290 295 300His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu Lys Gly Ala Val305 310 315 320Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly Tyr Ser Trp Glu
325 330 335Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu Leu Leu Ser Ser
340 345 350Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile Asp Ile Asn Asp
355 360 365Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His Tyr Ser Val Asp
370 375 380Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu Gln Lys Glu Leu385 390 395 400Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly Phe Ile Gly Arg
405 410 415Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu Ile Ile Pro Asp
420 425 430Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly Ser Gly Asp Pro
435 440 445Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile Phe Lys Asp Lys
450 455 460Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser His Arg Ile Thr465 470 475 480Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe Glu Pro Cys Gly
485 490 495Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val Pro Val Val His
500 505 510Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe Asn Pro Phe Gly
515 520 525Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala Pro Leu Thr Thr
530 535 540Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile Tyr Ile Gln Gly545 550 555 560Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg His Val Lys Arg
565 570 575Leu His Val Gly Pro Cys Arg ★
580(2)序列14资料:
(i)序列特征:
(A)长度:2725个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:mRNA
(iii)假说:无
(vi)来源:
(A)生物:玉米
(ix)特征:
(A)名称/键:sig-peptide
(B)位置:91..264
(ix)特征:
(A)名称/键:mat-peptide
(B)位置:265..2487
(ix)特征:
(A)名称/键:CDS
(B)位置:91..2490
(xi)序列描述:序列14:GGCCCAGAGC AGACCCGGAT TTCGCTCTTG CGGTCGCTGG GGTTTTAGCA TTGGCTGATC 60AGTTCGATCC GATCCGGCTG CGAAGGCGAG ATG GCG TTC CGG GTT TCT GGG GCG 114
Met Ala Phe Arg Val Ser Gly Ala
-58 -55GTG CTC GGT GGG GCC GTA AGG GCT CCC CGA CTC ACC GGC GGC GGG GAG 162Val Leu Gly Gly Ala Val Arg Ala Pro Arg Leu Thr Gly Gly Gly Glu-50 -45 -40 -35GGT AGT CTA GTC TTC CGG CAC ACC GGC CTC TTC TTA ACT CGG GGT GCT 210Gly Ser Leu Val Phe Arg His Thr Gly Leu Phe Leu Thr Arg Gly Ala
-30 -25 -20CGA GTT GGA TGT TCG GGG ACG CAC GGG GCC ATG CGC GCG GCG GCC GCG 258Arg Val Gly Cys Ser Gly Thr His Gly Ala Met Arg Ala Ala Ala Ala
-15 -10 -5GCC AGG AAG GCG GTC ATG GTT CCT GAG GGC GAG AAT GAT GGC CTC GCA 306Ala Arg Lys Ala Val Met Val Pro Glu Gly Glu Asn Asp Gly Leu Ala
1 5 10TCA AGG GCT GAC TCG GCT CAA TTC CAG TCG GAT GAA CTG GAG GTA CCA 354Ser Arg Ala Asp Ser Ala Gln Phe Gln Ser Asp Glu Leu Glu Val Pro15 20 25 30GAC ATT TCT GAA GAG ACA ACG TGC GGT GCT GGT GTG GCT GAT GCT CAA 402Asp Ile Ser Glu Glu Thr Thr Cys Gly Ala Gly Val Ala Asp Ala Gln
35 40 45GCC TTG AAC AGA GTT CGA GTG GTC CCC CCA CCA AGC GAT GGA CAA AAA 450Ala Leu Asn Arg Val Arg Val Val Pro Pro Pro Ser Asp Gly Gln Lys
50 55 60ATA TTC CAG ATT GAC CCC ATG TTG CAA GGC TAT AAG TAC CAT CTT GAG 498Ile Phe Gln Ile Asp Pro Met Leu Gln Gly Tyr Lys Tyr His Leu Glu
65 70 75TAT CGG TAC AGC CTC TAT AGA AGA ATC CGT TCA GAC ATT GAT GAA CAT 546Tyr Arg Tyr Ser Leu Tyr Arg Arg Ile Arg Ser Asp Ile Asp Glu His
80 85 90GAA GGA GGC TTG GAA GCC TTC TCC CGT AGT TAT GAG AAG TTT GGA TTT 594Glu Gly Gly Leu Glu Ala Phe Ser Arg Ser Tyr Glu Lys Phe Gly Phe95 100 105 110AAT GCC AGC GCG GAA GGT ATC ACA TAT CGA GAA TGG GCT CCT GGA GCA 642Asn Ala Ser Ala Glu Gly Ile Thr Tyr Arg Glu Trp Ala Pro Gly Ala
115 120 125TTT TCT GCA GCA TTG GTG GGT GAC GTC AAC AAC TGG GAT CCA AAT GCA 690Phe Ser Ala Ala Leu Val Gly Asp Val Asn Asn Trp Asp Pro Asn Ala
130 135 140GAT CGT ATG AGC AAA AAT GAG TTT GGT GTT TGG GAA ATT TTT CTG CCT 738Asp Arg Met Ser Lys Asn Glu Phe Gly Val Trp Glu Ile Phe Leu Pro
145 150 155AAC AAT GCA GAT GGT ACA TCA CCT ATT CCT CAT GGA TCT CGT GTA AAG 786Asn Asn Ala Asp Gly Thr Ser Pro Ile Pro His Gly Ser Arg Val Lys
160 165 170GTG AGA ATG GAT ACT CCA TCA GGG ATA AAG GAT TCA ATT CCA GCC TGG 834Val Arg Met Asp Thr Pro Ser Gly Ile Lys Asp Ser Ile Pro Ala Trp175 180 185 190ATC AAG TAC TCA GTG CAG GCC CCA GGA GAA ATA CCA TAT GAT GGG ATT 882Ile Lys Tyr Ser Val Gln Ala Pro Gly Glu Ile Pro Tyr Asp Gly Ile
195 200 205TAT TAT GAT CCT CCT GAA GAG GTA AAG TAT GTG TTC AGG CAT GCG CAA 930Tyr Tyr Asp Pro Pro Glu Glu Val Lys Tyr Val Phe Arg His Ala Gln
210 215 220CCT AAA CGA CCA AAA TCA TTG CGG ATA TAT GAA ACA CAT GTC GGA ATG 978Pro Lys Arg Pro Lys Ser Leu Arg Ile Tyr Glu Thr His Val Gly Met
225 230 235AGT AGC CCG GAA CCG AAG ATA AAC ACA TAT GTA AAC TTT AGG GAT GAA 1026Ser Ser Pro Glu Pro Lys Ile Asn Thr Tyr Val Asn Phe Arg Asp Glu
240 245 250GTC CTC CCA AGA ATA AAA AAA CTT GGA TAC AAT GCA GTG CAA ATA ATG 1074Val Leu Pro Arg Ile Lys Lys Leu Gly Tyr Asn Ala Val Gln Ile Met255 260 265 270CCA ATC CAA GAG CAC TCA TAT TAT GGA AGC TTT GGA TAC CAT GTA ACT 1122Ala Ile Gln Glu His Ser Tyr Tyr Gly Ser Phe Gly Tyr His Val Thr
275 280 285AAT TTT TTT GCG CCA AGT AGT CGT TTT GGT ACC CCA GAA GAT TTG AAG 1170Asn Phe Phe Ala Pro Ser Ser Arg Phe Gly Thr Pro Glu Asp Leu Lys
290 295 300TCT TTG ATT GAT AGA GCA CAT GAG CTT GGT TTG CTA GTT CTC ATG GAT 1218Ser Leu Ile Asp Arg Ala His Glu Leu Gly Leu Leu Val Leu Met Asp
305 310 315GTG GTT CAT AGT CAT GCG TCA AGT AAT ACT CTG GAT GGG TTG AAT GGT 1266Val Val His Ser His Ala Ser Ser Asn Thr Leu Asp Gly Leu Asn Gly
320 325 330TTT GAT GGT ACA GAT ACA CAT TAC TTT CAC AGT GGT CCA CGT GGC CAT 1314Phe Asp Gly Thr Asp Thr His Tyr Phe His Ser Gly Pro Arg Gly His335 340 345 350CAC TGG ATG TGG GAT TCT CGC CTA TTT AAC TAT GGG AAC TGG GAA GTT 1362His Trp Met Trp Asp Ser Arg Leu Phe Asn Tyr Gly Asn Trp Glu Val
355 360 365TTA AGA TTT CTT CTC TCC AAT GCT AGA TGG TGG CTC GAG GAA TAT AAG 1410Leu Arg Phe Leu Leu Ser Asn Ala Arg Trp Trp Leu Glu Glu Tyr Lys
370 375 380TTT GAT GGT TTC CGT TTT GAT GGT GTG ACC TCC ATG ATG TAC ACT CAC 1458Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser Met Met Tyr Thr His
385 390 395CAC GGA TTA CAA GTA ACA TTT ACG GGG AAC TTC AAT GAG TAT TTT GGC 1506His Gly Leu Gln Val Thr Phe Thr Gly Asn Phe Asn Glu Tyr Phe Gly
400 405 410TTT GCC ACC GAT GTA GAT GCA GTG GTT TAC TTG ATG CTG GTA AAT GAT 1554Phe Ala Thr Asp Val Asp Ala Val Val Tyr Leu Met Leu Val Asn Asp415 420 425 430CTA ATT CAT GGA CTT TAT CCT GAG GCT GTA ACC ATT GGT GAA GAT GTT 1602Leu Ile His Gly Leu Tyr Pro Glu Ala Val Thr Ile Gly Glu Asp Val
435 440 445AGT GGA ATG CCT ACA TTT GCC CTT CCT GTT CAC GAT GGT GGG GTA GGT 1650Ser Gly Met Pro Thr Phe Ala Leu Pro Val His Asp Gly Gly Val Gly
450 455 460TTT GAC TAT CGG ATG CAT ATG GCT GTG GCT GAC AAA TGG ATT GAC CTT 1698Phe Asp Tyr Arg Met His Met Ala Val Ala Asp Lys Trp Ile Asp Leu
465 470 475CTC AAG CAA AGT GAT GAA ACT TGG AAG ATG GGT GAT ATT GTG CAC ACA 1746Leu Lys Gln Ser Asp Glu Thr Trp Lys Met Gly Asp Ile Val His Thr
480 485 490CTG ACA AAT AGG AGG TGG TTA GAG AAG TGT GTA ACT TAT GCT GAA AGT 1794Leu Thr Asn Arg Arg Trp Leu Glu Lys Cys Val Thr Tyr Ala Glu Ser495 500 505 510CAT GAT CAA GCA TTA GTC GGC GAC AAG ACT ATT GCG TTT TGG TTG ATG 1842His Asp Gln Ala Leu Val Gly Asp Lys Thr Ile Ala Phe Trp Leu Met
515 520 525GAC AAG GAT ATG TAT GAT TTC ATG GCC CTC GAT AGA CCT TCA ACT CCT 1890Asp Lys Asp Met Tyr Asp Phe Met Ala Leu Asp Arg Pro Ser Thr Pro
530 535 540ACC ATT GAT CGT GGG ATA GCA TTA CAT AAG ATG ATT AGA CTT ATC ACA 1938Thr Ile Asp Arg Gly Ile Ala Leu His Lys Met Ile Arg Leu Ile Thr
545 550 555ATG GGT TTA GGA GGA GAG GGC TAT CTT AAT TTC ATG GGA AAT GAG TTT 1986Met Gly Leu Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn Glu Phe
560 565 570GGA CAT CCT GAA TGG ATA GAT TTT CCA AGA GGT CCG CAA AGA CTT CCA 2034Gly His Pro Glu Trp Ile Asp Phe Pro Arg Gly Pro Gln Arg Leu Pro575 580 585 590AGT GGT AAG TTT ATT CCA GGG AAT AAC AAC AGT TAT GAC AAA TGT CGT 2082Ser Gly Lys Phe Ile Pro Gly Asn Asn Asn Ser Tyr Asp Lys Cys Arg
595 600 605CGA AGA TTT GAC CTG GGT GAT GCA GAC TAT CTT AGG TAT CAT GGT ATG 2130Arg Arg Phe Asp Leu Gly Asp Ala Asp Tyr Leu Arg Tyr His Gly Met
610 615 620CAA GAG TTT GAT CAG GCA ATG CAA CAT CTT GAG CAA AAA TAT GAA TTC 2178Gln Glu Phe Asp Gln Ala Met Gln His Leu Glu Gln Lys Tyr Glu Phe
625 630 635ATG ACA TCT GAT CAC CAG TAT ATT TCC CGG AAA CAT GAG GAG GAT AAG 2226Met Thr Ser Asp His Gln Tyr Ile Ser Arg Lys His Glu Glu Asp Lys
640 645 650GTG ATT GTG TTC GAA AAG GGA GAT TTG GTA TTT GTG TTC AAC TTC CAC 2274Val Ile Val Phe Glu Lys Gly Asp Leu Val Phe Val Phe Asn Phe His655 660 665 670TGC AAC AAC AGC TAT TTT GAC TAC CGT ATT GGT TGT CGA AAG CCT GGG 2322Cys Asn Asn Ser Tyr Phe Asp Tyr Arg Ile Gly Cys Arg Lys Pro Gly
675 680 685GTG TAT AAG GTG GTC TTG GAC TCC GAC GCT GGA CTA TTT GGT GGA TTT 2370Val Tyr Lys Val Val Leu Asp Ser Asp Ala Gly Leu Phe Gly Gly Phe
690 695 700AGC AGG ATC CAT CAC GCA GCC GAG CAC TTC ACC GCC GAC TGT TCG CAT 2418Ser Arg Ile His His Ala Ala Glu His Phe Thr Ala Asp Cys Ser His
705 710 715GAT AAT AGG CCA TAT TCA TTC TCG GTT TAT ACA CCA AGC AGA ACA TGT 2466Asp Asn Arg Pro Tyr Ser Phe Ser Val Tyr Thr Pro Ser Arg Thr Cys
720 725 730GTC GTC TAT GCT CCA GTG GAG TGA TAGCGGGGTA CTCGTTGCTG CGCGGCATGT 2520Val Val Tyr Ala Pro Val Glu ★735 740GTGGGGCTGT CGATGTGAGG AAAAACCTTC TTCCAAAACC GGCAGATGCA TGCATGCATG 2580CTACAATAAG GTTCTGATAC TTTAATCGAT GCTGGAAAGC CCATGCATCT CGCTGCGTTG 2640TCCTCTCTAT ATATATAAGA CCTTCAAGGT GTCAATTAAA CATAGAGTTT TCGTTTTTCG 2700CTTTCCTAAA AAAAAAAAAA AAA 2725(2)序列15资料:
(i)序列特征:
(A)长度:800氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列15:Met Ala Phe Arg Val Ser Gly Ala Val Leu Gly Gly Ala Val Arg Ala-58 -55 -50 -45Pro Arg Leu Thr Gly Gly Gly Glu Gly Ser Leu Val Phe Arg His Thr
-40 -35 -30Gly Leu Phe Leu Thr Arg Gly Ala Arg Val Gly Cys Ser Gly Thr His
-25 -20 -15Gly Ala Met Arg Ala Ala Ala Ala Ala Arg Lys Ala Val Met Val Pro-10 -5 1 5Glu Gly Glu Asn Asp Gly Leu Ala Ser Arg Ala Asp Ser Ala Gln Phe
10 15 20Gln Ser Asp Glu Leu Glu Val Pro Asp Ile Ser Glu Glu Thr Thr Cys
25 30 35Gly Ala Gly Val Ala Asp Ala Gln Ala Leu Asn Arg Val Arg Val Val
40 45 50Pro Pro Pro Ser Asp Gly Gln Lys Ile Phe Gln Ile Asp Pro Met Leu55 60 65 70Gln Gly Tyr Lys Tyr His Leu Glu Tyr Arg Tyr Ser Leu Tyr Arg Arg
75 80 85Ile Arg Ser Asp Ile Asp Glu His Glu Gly Gly Leu Glu Ala Phe Ser
90 95 100Arg Ser Tyr Glu Lys Phe Gly Phe Asn Ala Ser Ala Glu Gly Ile Thr
105 110 115Tyr Arg Glu Trp Ala Pro Gly Ala Phe Ser Ala Ala Leu Val Gly Asp
120 125 130Val Asn Asn Trp Asp Pro Asn Ala Asp Arg Met Ser Lys Asn Glu Phe135 140 145 150Gly Val Trp Glu Ile Phe Leu Pro Asn Asn Ala Asp Gly Thr Ser Pro
155 160 165Ile Pro His Gly Ser ArgVal Lys Val Arg Met Asp Thr Pro Ser Gly
170 175 180Ile Lys Asp Ser Ile Pro Ala Trp Ile Lys Tyr Ser Val Gln Ala Pro
185 190 195Gly Glu Ile Pro Tyr Asp Gly Ile Tyr Tyr Asp Pro Pro Glu Glu Val
200 205 210Lys Tyr Val Phe Arg His Ala Gln Pro Lys Arg Pro Lys Ser Leu Arg215 220 225 230Ile Tyr Glu Thr His Val Gly Met Ser Ser Pro Glu Pro Lys Ile Asn
235 240 245Thr Tyr Val Asn Phe Arg Asp Glu Val Leu Pro Arg Ile Lys Lys Leu
250 255 260Gly Tyr Asn Ala Val Gln Ile Met Ala Ile Gln Glu His Ser Tyr Tyr
265 270 275Gly Ser Phe Gly Tyr His Val Thr Asn Phe Phe Ala Pro Ser Ser Arg
280 285 290Phe Gly Thr Pro Glu Asp Leu Lys Ser Leu Ile Asp Arg Ala is Glu295 300 305 310Leu Gly Leu Leu Val Leu Met Asp Val Val His Ser His Ala Ser Ser
315 320 325Asn Thr Leu Asp Gly Leu Asn Gly Phe Asp Gly Thr Asp Thr His Tyr
330 335 340Phe Hie Ser Gly Pro Arg Gly His His Trp Met Trp Asp Ser Arg Leu
345 350 355Phe Asn Tyr Gly Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Ala
360 365 370Arg Trp Trp Leu Glu Glu Tyr Lys Phe Asp Gly Phe Arg Phe Asp Gly375 380 385 390Val Thr Ser Met Met Tyr Thr His His Gly Leu Gln Val Thr Phe Thr
395 400 405Gly Asn Phe Asn Glu Tyr Phe Gly Phe Ala Thr Asp Val Asp Ala Val
410 415 420Val Tyr Leu Met Leu Val Asn Asp Leu Ile His Gly Leu Tyr Pro Glu
425 430 435Ala Val Thr Ile Gly Glu Asp Val Ser Gly Met Pro Thr Phe Ala Leu
440 445 450Pro Val His Asp Gly Gly Val Gly Phe Asp Tyr Arg Met His Met Ala455 460 465 470Val Ala Asp Lys Trp Ile Asp Leu Leu Lys Gln Ser Asp Glu Thr Trp
475 480 485Lys Met Gly Asp Ile Val His Thr Leu Thr Asn Arg Arg Trp Leu Glu
490 495 500Lys Cys Val Thr Tyr Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp
505 510 515Lys Thr Ile Ala Phe Trp Leu Met Asp Lys Asp Met Tyr Asp Phe Met
520 525 530Ala Leu Asp Arg Pro Ser Thr Pro Thr Ile Asp Arg Gly Ile Ala Leu535 540 545 550His Lys Met Ile Arg Leu Ile Thr Met Gly Leu Gly Gly Glu Gly Tyr
555 560 565Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe
570 575 580Pro Arg Gly Pro Gln Arg Leu Pro Ser Gly Lys Phe Ile Pro Gly Asn
585 590 595Asn Asn Ser Tyr Asp Lys Cys Arg Arg Arg Phe Asp Leu Gly Asp Ala
600 605 610Asp Tyr Leu Arg Tyr His Gly Met Gln Glu Phe Asp Gln Ala Met Gln615 620 625 630His Leu Glu Gln Lys Tyr Glu Phe Met Thr Ser Asp His Gln Tyr Ile
635 640 645Ser Arg Lys His Glu Glu Asp Lys Val Ile Val Phe Glu Lys Gly Asp
650 655 660Leu Val Phe Val Phe Asn Phe His Cys Asn Asn Ser Tyr Phe Asp Tyr
665 670 675Arg Ile Gly Cys Arg Lys Pro Gly Val Tyr Lys Val Val Leu Asp Ser
680 685 690Asp Ala Gly Leu Phe Gly Gly Phe Ser Arg Ile His His Ala Ala Glu695 700 705 710His Phe Thr Ala Asp Cys Ser His Asp Asn Arg Pro Tyr Ser Phe Ser
715 720 725Val Tyr Thr Pro Ser Arg Thr Cys Val Val Tyr Ala Pro Val Glu ★
730 735 740(2)序列16资料:
(i)序列特征:
(A)长度:2763个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:mRNA
(iii)假说:无
(vi)来源:
(A)生物:玉米
(ix)特征:
(A)名称/键:transit-peptide
(B)位置:2..190
(ix)特征:
(A)名称/键:mat-peptide
(B)位置:191..2467
(ix)特征:
(A)名称/键:CDS
(B)位置:2..2470
(xi)序列描述:序列16:G CTG TGC CTC GTG TCG CCC TCT TCC TCG CCG ACT CCG CTT CCG CCG 46Leu Cys Leu Val Ser Pro Ser Ser Ser Pro Thr Pro Leu Pro Pro-63 -60 -55 -50CCG CGG CGC TCT CGC TCG CAT GCT GAT CGG GCG GCA CCG CCG GGG ATC 94Pro Arg Arg Ser Arg Ser His Ala Asp Arg Ala Ala Pro Pro Gly Ile
-45 -40 -35GCG GGT GGC GGC AAT GTG CGC CTG AGT GTG TTG TCT GTC CAG TGC AAG 142Ala Gly Gly Gly Asn Val Arg Leu Ser Val Leu Ser Val Gln Cys Lys
-30 -25 -20GCT CGC CGG TCA GGG GTG CGG AAG GTC AAG AGC AAA TTC GCC ACT GCA 190Ala Arg Arg Ser Gly Val Arg Lys Val Lys Ser Lys Phe Ala Thr Ala
-15 -10 -5GCT ACT GTG CAA GAA GAT AAA ACT ATG GCA ACT GCC AAA GGC GAT GTC 238Ala Thr Val Gln Glu Asp Lys Thr Met Ala Thr Ala Lys Gly Asp Val1 5 10 15GAC CAT CTC CCC ATA TAC GAC CTG GAC CCC AAG CTG GAG ATA TTC AAG 286Asp His Leu Pro Ile Tyr Asp Leu Asp Pro Lys Leu Glu Ile Phe Lys
20 25 30GAC CAT TTC AGG TAC CGG ATG AAA AGA TTC CTA GAG CAG AAA GGA TCA 334Asp His Phe Arg Tyr Arg Met Lys Arg Phe Leu Glu Gln Lys Gly Ser
35 40 45ATT GAA GAA AAT GAG GGA AGT CTT GAA TCT TTT TCT AAA GGC TAT TTG 382Ile Glu Glu Asn Glu Gly Ser Leu Glu Ser Phe Ser Lys Gly Tyr Leu
50 55 60AAA TTT GGG ATT AAT ACA AAT GAG GAT GGA ACT GTA TAT CGT GAA TGG 430Lys Phe Gly Ile Asn Thr Asn Glu Asp Gly Thr Val Tyr Arg Glu Trp65 70 75 80GCA CCT GCT GCG CAG GAG GCA GAG CTT ATT GGT GAC TTC AAT GAC TGG 478Ala Pro Ala Ala Gln Glu Ala Glu Leu Ile Gly Asp Phe Asn Asp Trp
85 90 95AAT GGT GCA AAC CAT AAG ATG GAG AAG GAT AAA TTT GGT GTT TGG TCG 526Asn Gly Ala Asn His Lys Met Glu Lys Asp Lys Phe Gly Val Trp Ser
100 105 110ATC AAA ATT GAC CAT GTC AAA GGG AAA CCT GCC ATC CCT CAC AAT TCC 574Ile Lys Ile Asp His Val Lys Gly Lys Pro Ala Ile Pro His Asn Ser
115 120 125AAG GTT AAA TTT CGC TTT CTA CAT GGT GGA GTA TGG GTT GAT CGT ATT 622Lys Val Lys Phe Arg Phe Leu His Gly Gly Val Trp Val Asp Arg Ile
130 135 140CCA GCA TTG ATT CGT TAT GCG ACT GTT GAT GCC TCT AAA TTT GGA GCT 670Pro Ala Leu Ile Arg Tyr Ala Thr Val Asp Ala Ser Lys Phe Gly Ala145 150 155 160CCC TAT GAT GGT GTT CAT TGG GAT CCT CCT GCT TCT GAA AGG TAC ACA 718Pro Tyr Asp Gly Val His Trp Asp Pro Pro Ala Ser Glu Arg Tyr Thr
165 170 175TTT AAG CAT CCT CGG CCT TCA AAG CCT GCT GCT CCA CGT ATC TAT GAA 766Phe Lys His Pro Arg Pro Ser Lys Pro Ala Ala Pro Arg Ile Tyr Glu
180 185 190GCC CAT GTA GGT ATG AGT GGT GAA AAG CCA GCA GTA AGC ACA TAT AGG 814Ala His Val Gly Met Ser Gly Glu Lys Pro Ala Val Ser Thr Tyr Arg
195 200 205GAA TTT GCA GAC AAT GTG TTG CCA CGC ATA CGA GCA AAT AAC TAC AAC 862Glu Phe Ala Asp Asn Val Leu Pro Arg Ile Arg Ala Asn Asn Tyr Asn
210 215 220ACA GTT CAG TTG ATG GCA GTT ATG GAG CAT TCG TAC TAT GCT TCT TTC 910
116Thr Val Gln Leu Met Ala Val Met Glu His Ser Tyr Tyr Ala Ser Phe225 230 235 240GGG TAC CAT GTG ACA AAT TTC TTT GCG GTT AGC AGC AGA TCA GGC ACA 958Gly Tyr His Val Thr Asn Phe Phe Ala Val Ser Ser Arg Ser Gly Thr
245 250 255CCA GAG GAC CTC AAA TAT CTT GTT GAT AAG GCA CAC AGT TTG GGT TTG 1006Pro Glu Asp Leu Lys Tyr Leu Val Asp Lys Ala His Ser Leu Gly Leu
260 265 270CGA GTT CTG ATG GAT GTT GTC CAT AGC CAT GCA AGT AAT AAT GTC ACA 1054Arg Val Leu Met Asp Val Val His Ser His Ala Ser Asn Asn Val Thr
275 280 285GAT GGT TTA AAT GGC TAT GAT GTT GGA CAA AGC ACC CAA GAG TCC TAT 1102Asp Gly Leu Asn Gly Tyr Asp Val Gly Gln Ser Thr Gln Glu Ser Tyr
290 295 300TTT CAT GCG GGA GAT AGA GGT TAT CAT AAA CTT TGG GAT AGT CGG CTG 1150Phe His Ala Gly Asp Arg Gly Tyr His Lys Leu Trp Asp Ser Arg Leu305 310 315 320TTC AAC TAT GCT AAC TGG GAG GTA TTA AGG TTT CTT CTT TCT AAC CTG 1198Phe Asn Tyr Ala Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Leu
325 330 335AGA TAT TGG TTG GAT GAA TTC ATG TTT GAT GGC TTC CGA TTT GAT GGA 1246Arg Tyr Trp Leu Asp Glu Phe Met Phe Asp Gly Phe Arg Phe Asp Gly
340 345 350GTT ACA TCA ATG CTG TAT CAT CAC CAT GGT ATC AAT GTG GGG TTT ACT 1294Val Thr Ser Met Leu Tyr His His His Gly Ile Asn Val Gly Phe Thr
355 360 365GGA AAC TAC CAG GAA TAT TTC AGT TTG GAC ACA GCT GTG GAT GCA GTT 1342Gly Asn Tyr Gln Glu Tyr Phe Ser Leu Asp Thr Ala Val Asp Ala Val
370 375 380GTT TAC ATG ATG CTT GCA AAC CAT TTA ATG CAC AAA CTC TTG CCA GAA 1390Val Tyr Met Met Leu Ala Asn His Leu Met His Lys Leu Leu Pro Glu385 390 395GCA ACT GTT GTT GCT GAA GAT GTT TCA GGC ATG CCG GTC CTT TGC CGG 1438Ala Thr Val Val Ala Glu Asp Val Ser Gly Met Pro Val Leu Cys Arg
405 410 415CCA GTT GAT GAA GGT GGG GTT GGG TTT GAC TAT CGC CTG GCA ATG GCT 1486Pro Val Asp Glu Gly Gly Val Gly Phe Asp Tyr Arg Leu Ala Met Ala
420 425 430ATC CCT GAT AGA TGG ATT GAC TAC CTG AAG AAT AAA GAT GAC TCT GAG 1534Ile Pro Asp Arg Trp Ile Asp Tyr Leu Lys Asn Lys Asp Asp Ser Glu
435 440 445TGG TCG ATG GGT GAA ATA GCG CAT ACT TTG ACT AAC AGG AGA TAT ACT 1582Trp Ser Met Gly Glu Ile Ala His Thr Leu Thr Asn Arg Arg Tyr Thr
450 455 460GAA AAA TGC ATC GCA TAT GCT GAG AGC CAT GAT CAG TCT ATT GTT GGC 1630Glu Lys Cys Ile Ala Tyr Ala Glu Ser His Asp Gln Ser Ile Val Gly465 470 475 480GAC AAA ACT ATT GCA TTT CTC CTG ATG GAC AAG GAA ATG TAC ACT GGC 1678Asp Lys Thr Ile Ala Phe Leu Leu Met Asp Lys Glu Met Tyr Thr Gly
485 490 495ATG TCA GAC TTG CAG CCT GCT TCA CCT ACA ATT GAT CGA GGG ATT GCA 1726Met Ser Asp Leu Gln Pro Ala Ser Pro Thr Ile Asp Arg Gly Ile Ala
500 505 510CTC CAA AAG ATG ATT CAC TTC ATC ACA ATG GCC CTT GGA GGT GAT GGC 1774Leu Gln Lys Met Ile His Phe Ile Thr Met Ala Leu Gly Gly Asp Gly
515 520 525TAC TTG AAT TTT ATG GGA AAT GAG TTT GGT CAC CCA GAA TGG ATT GAC 1822Tyr Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp
530 535 540TTT CCA AGA GAA GGG AAC AAC TGG AGC TAT GAT AAA TGC AGA CGA CAG 1870Phe Pro Arg Glu Gly Asn Asn Trp Ser Tyr Asp Lys Cys Arg Arg Gln545 550 555 560TGG AGC CTT GTG GAC ACT GAT CAC TTG CGG TAC AAG TAC ATG AAT GCG 1918Trp Ser Leu Val Asp Thr Asp His Leu Arg Tyr Lys Tyr Met Asn Ala
565 570 575TTT GAC CAA GCG ATG AAT GCG CTC GAT GAG AGA TTT TCC TTC CTT TCG 1966Phe Asp Gln Ala Met Asn Ala Leu Asp Glu Arg Phe Ser Phe Leu Ser
580 585 590TCG TCA AAG CAG ATC GTC AGC GAC ATG AAC GAT GAG GAA AAG GTT ATT 2014Ser Ser Lys Gln Ile Val Ser Asp Met Asn Asp Glu Glu Lys Val Ile
595 600 605GTC TTT GAA CGT GGA GAT TTA GTT TTT GTT TTC AAT TTC CAT CCC AAG 2062Val Phe Glu Arg Gly Asp Leu Val Phe Val Phe Asn Phe His Pro Lys
610 615 620AAA ACT TAC GAG GGC TAC AAA GTG GGA TGC GAT TTG CCT GGG AAA TAC 2110Lys Thr Tyr Glu Gly Tyr Lys Val Gly Cys Asp Leu Pro Gly Lys Tyr625 630 635 640AGA GTA GCC CTG GAC TCT GAT GCT CTG GTC TTC GGT GGA CAT GGA AGA 2158Arg Val Ala Leu Asp Ser Asp Ala Leu Val Phe Gly Gly His Gly Arg
645 650 655GTT GGC CAC GAC GTG GAT CAC TTC ACG TCG CCT GAA GGG GTG CCA GGG 2206Val Gly His Asp Val Asp His Phe Thr Ser Pro Glu Gly Val Pro Gly
660 665 670GTG CCC GAA ACG AAC TTC AAC AAC CGG CCG AAC TCG TTC AAA GTC CTT 2254Val Pro Glu Thr Asn Phe Asn Asn Arg Pro Asn Ser Phe Lys Val Leu
675 680 685TCT CCG CCC CGC ACC TGT GTG GCT TAT TAC CGT GTA GAC GAA GCA GGG 2302Ser Pro Pro Arg Thr Cys Val Ala Tyr Tyr Arg Val Asp Glu Ala Gly
690 695 700GCT GGA CGA CGT CTT CAC GCG AAA GCA GAG ACA GGA AAG ACG TCT CCA 2350Ala Gly Arg Arg Leu His Ala Lys Ala Glu Thr Gly Lys Thr Ser Pro705 710 715 720GCA GAG AGC ATC GAC GTC AAA GCT TCC AGA GCT AGT AGC AAA GAA GAC 2398Ala Glu Ser Ile Asp Val Lys Ala Ser Arg Ala Ser Ser Lys Glu Asp
725 730 735AAG GAG GCA ACG GCT GGT GGC AAG AAG GGA TGG AAG TTT GCG CGG CAG 2446Lys Glu Ala Thr Ala Gly Gly Lys Lys Gly Trp Lys Phe Ala Arg Gln
740 745 750CCA TCC GAT CAA GAT ACC AAA TGA AGCCACGAGT CCTTGGTGAG GACTGGACTG 2500Pro Ser Asp Gln Asp Thr Lys ★
755 760GCTGCCGGCG CCCTGTTAGT AGTCCTGCTC TACTGGACTA GCCGCCGCTG GCGCCCTTGG 2560AACGGTCCTT TCCTGTAGCT TGCAGGCGAC TGGTGTCTCA TCACCGAGCA GGCAGGCACT 2620GCTTGTATAG CTTTTCTAGA ATAATAATCA GGGATGGATG GATGGTGTGT ATTGGCTATC 2680TGGCTAGACG TGCATGTGCC CAGTTTGTAT GTACAGGAGC AGTTCCCGTC CAGAATAAAA 2740AAAAACGT TGGGGGGTTT TTC 2763(2)序列17资料:
(i)序列特征:
(A)长度:823个氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列17:Leu Cys Leu Val Ser Pro Ser Ser Ser Pro Thr Pro Leu Pro Pro Pro-63 -60 -55 -50Arg Arg Ser Arg Ser His Ala Asp Arg Ala Ala Pro Pro Gly Ile Ala
-45 -40 -35Gly Gly Gly Asn Val Arg Leu Ser Val Leu Ser Val Gln Cys Lys Ala
-30 -25 -20Arg Arg Ser Gly Val Arg Lys Val Lys Ser Lys Phe Ala Thr Ala Ala-15 -10 -5 1Thr Val Gln Glu Asp Lys Thr Met Ala Thr Ala Lys Gly Asp Val Asp
5 10 15His Leu Pro Ile Tyr Asp Leu Asp Pro Lys Leu Glu Ile Phe Lys Asp
20 25 30His Phe Arg Tyr Arg Met Lys Arg Phe Leu Glu Gln Lys Gly Ser Ile
35 40 45Glu Glu Asn Glu Gly Ser Leu Glu Ser Phe Ser Lys Gly Tyr Leu Lys50 55 60 65Phe Gly Ile Asn Thr Asn Glu Asp Gly Thr Val Tyr Arg Glu Trp Ala
70 75 80Pro Ala Ala Gln Glu Ala Glu Leu Ile Gly Asp Phe Asn Asp Trp Asn
85 90 95Gly Ala Asn His Lys Met Glu Lys Asp Lys Phe Gly Val Trp Ser Ile
100 105 110Lys Ile Asp His Val Lys Gly Lys Pro Ala Ile Pro His Asn Ser Lys
115 120 125Val Lys Phe Arg Phe Leu His Gly Gly Val Trp Val Asp Arg Ile Pro130 135 140 145Ala Leu Ile Arg Tyr Ala Thr Val Asp Ala Ser Lys Phe Gly Ala Pro
150 155 160Tyr Asp Gly Val His Trp Asp Pro Pro Ala Ser Glu Arg Tyr Thr Phe
165 170 175Lys His Pro Arg Pro Ser Lys Pro Ala Ala Pro Arg Ile Tyr Glu Ala
180 185 190His Val Gly Met Ser Gly Glu Lys Pro Ala Val Ser Thr Tyr Arg Glu
195 200 205Phe Ala Asp Asn Val Leu Pro Arg Ile Arg Ala Asn Asn Tyr Asn Thr210 215 220Val Gln Leu Met Ala Val Met Glu His Ser Tyr Tyr Ala Ser Phe Gly
230 235 240Tyr His Val Thr Asn Phe Phe Ala Val Ser Ser Arg Ser Gly Thr Pro
245 250 255Glu Asp Leu Lys Tyr Leu Val Asp Lys Ala His Ser Leu Gly Leu Arg
260 265 270Val Leu Met Asp Val Val His Ser His Ala Ser Asn Asn Val Thr Asp
275 280 285Gly Leu Asn Gly Tyr Asp Val Gly Gln Ser Thr Gln Glu Ser Tyr Phe290 295 300 305His Ala Gly Asp Arg Gly Tyr His Lys Leu Trp Asp Ser Arg Leu Phe
310 315 320Asn Tyr Ala Asn Trp Glu Val Leu Arg Phe Leu Leu Ser Asn Leu Arg
325 330 335Tyr Trp Leu Asp Glu Phe Met Phe Asp Gly Phe Arg Phe Asp Gly Val
340 345 350Thr Ser Met Leu Tyr His His His Gly Ile Asn Val Gly Phe Thr Gly
355 360 365Asn Tyr Gln Glu Tyr Phe Ser Leu Asp Thr Ala Val Asp Ala Val Val370 375 380 385Tyr Met Met Leu Ala Asn His Leu Met His Lys Leu Leu Pro Glu Ala
390 395 400Thr Val Val Ala Glu Asp Val Ser Gly Met Pro Val Leu Cys Arg Pro
405 410 415Val Asp Glu Gly Gly Val Gly Phe Asp Tyr Arg Leu Ala Met Ala Ile
420 425 430Pro Asp Arg Trp Ile Asp Tyr Leu Lys Asn Lys Asp Asp Ser Glu Trp
435 440 445Ser Met Gly Glu Ile Ala His Thr Leu Thr Asn Arg Arg Tyr Thr Glu450 455 460 465Lys Cys Ile Ala Tyr Ala Glu Ser His Asp Gln Ser Ile Val Gly Asp
470 475 480Lys Thr Ile Ala Phe Leu Leu Met Asp Lys Glu Met Tyr Thr Gly Met
485 490 495Ser Asp Leu Gln Pro Ala Ser Pro Thr Ile Asp Arg Gly Ile Ala Leu
500 505 510Gln Lys Met Ile His Phe Ile Thr Met Ala Leu Gly Gly Asp Gly Tyr
515 520 525Leu Asn Phe Met Gly Asn Glu Phe Gly His Pro Glu Trp Ile Asp Phe530 535 540 545Pro Arg Glu Gly Asn Asn Trp Ser Tyr Asp Lys Cys Arg Arg Gln Trp
550 555 560Ser Leu Va1 Asp Thr Asp His Leu Arg Tyr Lys Tyr Met Asn Ala Phe
565 570 575Asp Gln Ala Met Asn Ala Leu Asp Glu Arg Phe Ser Phe Leu Ser Ser
580 585 590Ser Lys Gln Ile Val Ser Asp Met Asn Asp Glu Glu Lys Val Ile Val
595 600 605Phe Glu Arg Gly Asp Leu Val Phe Val Phe Asn Phe His Pro Lys Lys610 615 620 625Thr Tyr Glu Gly Tyr Lys Val Gly Cys Asp Leu Pro Gly Lys Tyr Arg
630 635 640Val Ala Leu Asp Ser Asp Ala Leu Val Phe Gly Gly Mis Gly Arg Val
645 650 655Gly His Asp Val Asp His Phe Thr Ser Pro Glu Gly Val Pro Gly Val
660 665 670Pro Glu Thr Asn Phe Asn Asn Arg Pro Asn Ser Phe Lys Val Leu Ser
675 680 685Pro Pro Arg Thr Cys Val Ala Tyr Tyr Arg Val Asp Glu Ala Gly Ala690 695 700 705Gly Arg Arg Leu His Ala Lys Ala Glu Thr Gly Lys Thr Ser Pro Ala
710 715 720Glu Ser Ile Asp Val Lys Ala Ser Arg Ala Ser Ser Lys Glu Asp Lys
725 730 735Glu Ala Thr Ala Gly Gly Lys Lys Gly Trp Lys Phe Ala Arg Gln Pro
740 745 750Ser Asp Gln Asp Thr Lys ★
755 760(2)序列18资料:
(i)序列特征:
(A)长度:153个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(vi)来源:
(A)生物:玉米
(ix)特征:
(A)名称/键:CDS
(B)位置:1..153
(xi)序列描述:序列18:ATG GCG ACG CCC TCG GCC GTG GGC GCC GCG TGC CTC CTC CTC GCG CGG 48Met Ala Thr Pro Ser Ala Val Gly Ala Ala Cys Leu Leu Leu Ala Arg
765 770 775GCC GCC TGG CCG GCC GCC GTC GGC GAC CGG GCG CGC CCG CGG AGG CTC 96Ala Ala Trp Pro Ala Ala Val Gly Asp Arg Ala Arg Pro Arg Arg Leu
780 785 790CAG CGC GTG CTG CGC CGC CGG TGC GTC GCG GAG CTG AGC AGG GAG GGG 144Gln Arg Val Leu Arg Arg Arg Cys Val Ala Glu Leu Ser Arg Glu Gly
795 800 805CCC CAT ATG 153Pro His Met
810(2)序列19资料:
(i)序列特征:
(A)长度:51个氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列19:Met Ala Thr Pro Ser Ala Val Gly Ala Ala Cys Leu Leu Leu Ala Arg1 5 10 15Ala Ala Trp Pro Ala Ala Val Gly Asp Arg Ala Arg Pro Arg Arg Leu
20 25 30Gln Arg Val Leu Arg Arg Arg Cys Val Ala Glu Leu Ser Arg Glu Gly
35 40 45Pro His Met
50(2)序列20资料:
(i)序列特征:
(A)长度:1620个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(i x)特征:
(A)名称/键:CDS
(B)位置:1..1620
(xi)序列描述:序列20:TGC GTC GCG GAG CTG AGC AGG GAG GAC CTC GGT CTC GAA CCT GAA GGG 48Cys Val Ala Glu Leu Ser Arg Glu Asp Leu Gly Leu Glu Pro Glu Gly
55 60 65ATT GCT GAA GGT TCC ATC GAT AAC ACA GTA GTT GTG GCA AGT GAG CAA 96Ile Ala Glu Gly Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln
70 75 80GAT TCT GAG ATT GTG GTT GGA AAG GAG CAA GCT CGA GCT AAA GTA ACA 144Asp Ser Glu Ile Val Val Gly Lys Glu Gln Ala Arg Ala Lys Val Thr
85 90 95CAA AGC ATT GTC TTT GTA ACC GGC GAA GCT TCT CCT TAT GCA AAG TCT 192Gln Ser Ile Val Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser100 105 110 115GGG GGT CTA GGA GAT GTT TGT GGT TCA TTG CCA GTT GCT CTT GCT GCT 240Gly Gly Leu Gly Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala
120 125 130CGT GGT CAC CGT GTG ATG GTT GTA ATG CCC AGA TAT TTA AAT GGT ACC 288Arg Gly His Arg Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr
135 140 145TCC GAT AAG AAT TAT GCA AAT GCA TTT TAC ACA GAA AAA CAC ATT CGG 336Ser Asp Lys Asn Tyr Ala Asn Ala Phe Tyr Thr Glu Lys His Ile Arg
150 155 160ATT CCA TGC TTT GGC GGT GAA CAT GAA GTT ACC TTC TTC CAT GAG TAT 384Ile Pro Cys Phe Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr
165 170 175AGA GAT TCA GTT GAC TGG GTG TTT GTT GAT CAT CCC TCA TAT CAC AGA 432Arg Asp Ser Val Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg180 185 190 195CCT GGA AAT TTA TAT GGA GAT AAG TTT GGT GCT TTT GGT GAT AAT CAG 480Pro Gly Asn Leu Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln
200 205 210TTC AGA TAC ACA CTC CTT TGC TAT GCT GCA TGT GAG GCT CCT TTG ATC 528Phe Arg Tyr Thr Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile
215 220 225CTT GAA TTG GGA GGA TAT ATT TAT GGA CAG AAT TGC ATG TTT GTT GTC 576Leu Glu Leu Gly Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val
230 235 240AAT GAT TGG CAT GCC AGT CTA GTG CCA GTC CTT CTT GCT GCA AAA TAT 624Asn Asp Trp His Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr
245 250 255AGA CCA TAT GGT GTT TAT AAA GAC TCC CGC AGC ATT CTT GTA ATA CAT 672Arg Pro Tyr Gly Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile His260 265 270 275AAT TTA GCA CAT CAG GGT GTA GAG CCT GCA AGC ACA TAT CCT GAC CTT 720Asn Leu Ala His Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu
280 285 290GGG TTG CCA CCT GAA TGG TAT GGA GCT CTG GAG TGG GTA TTC CCT GAA 768Gly Leu Pro Pro Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu
295 300 305TGG GCG AGG AGG CAT GCC CTT GAC AAG GGT GAG GCA GTT AAT TTT TTG 816Trp Ala Arg Arg His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu
310 315 320AAA GGT GCA GTT GTG ACA GCA GAT CGA ATC GTG ACT GTC AGT AAG GGT 864Lys Gly Ala Val Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly
325 330 335TAT TCG TGG GAG GTC ACA ACT GCT GAA GGT GGA CAG GGC CTC AAT GAG 912Tyr Ser Trp Glu Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu340 345 350 355CTC TTA AGC TCC AGA AAG AGT GTA TTA AAC GGA ATT GTA AAT GGA ATT 960Leu Leu Ser Ser Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile
360 365 370GAC ATT AAT GAT TGG AAC CCT GCC ACA GAC AAA TGT ATC CCC TGT CAT 1008Asp Ile Asn Asp Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His
375 380 385TAT TCT GTT GAT GAC CTC TCT GGA AAG GCC AAA TGT AAA GGT GCA TTG 1056Tyr Ser Val Asp Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu
390 395 400CAG AAG GAG CTG GGT TTA CCT ATA AGG CCT GAT GTT CCT CTG ATT GGC 1104Gln Lys Glu Leu Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly
405 410 415TTT ATT GGA AGG TTG GAT TAT CAG AAA GGC ATT GAT CTC ATT CAA CTT 1152Phe Ile Gly Arg Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu420 425 430 435ATC ATA CCA GAT CTC ATG CGG GAA GAT GTT CAA TTT GTC ATG CTT GGA 1200Ile Ile Pro Asp Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly
440 445 450TCT GGT GAC CCA GAG CTT GAA GAT TGG ATG AGA TCT ACA GAG TCG ATC 1248Ser Gly Asp Pro Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile
455 460 465TTC AAG GAT AAA TTT CGT GGA TGG GTT GGA TTT AGT GTT CCA GTT TCC 1296Phe Lys Asp Lys Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser
470 475 480CAC CGA ATA ACT GCC GGC TGC GAT ATA TTG TTA ATG CCA TCC AGA TTC 1344His Arg Ile Thr Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe
485 490 495GAA CCT TGT GGT CTC AAT CAG CTA TAT GCT ATG CAG TAT GGC ACA GTT 1392Glu Pro Cys Gly Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val500 505 510 515CCT GTT GTC CAT GCA ACT GGG GGC CTT AGA GAT ACC GTG GAG AAC TTC 1440Pro Val Val His Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe
520 525 530AAC CCT TTC GGT GAG AAT GGA GAG CAG GGT ACA GGG TGG GCA TTC GCA 1488Asn Pro Phe Gly Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala
535 540 545CCC CTA ACC ACA GAA AAC ATG TTT GTG GAC ATT GCG AAC TGC AAT ATC 1536Pro Leu Thr Thr Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile
550 555 560TAC ATA CAG GGA ACA CAA GTC CTC CTG GGA AGG GCT AAT GAA GCG AGG 1584Tyr Ile Gln Gly Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg
565 570 575CAT GTC AAA AGA CTT CAC GTG GGA CCA TGC CGC TGA 1620His Val Lys Arg Leu His Val Gly Pro Cys Arg ★580 585 590(2)序列21资料:
(i)序列特征:
(A)长度:540个氨基酸
(B)类型:氨基酸
(D)拓扑结构:线形
(ii)分子类型:蛋白
(xi)序列描述:序列21:Cys Val Ala Glu Leu Ser Arg Glu Asp Leu Gly Leu Glu Pro Glu Gly 1 5 10 15Ile Ala Glu Gly Ser Ile Asp Asn Thr Val Val Val Ala Ser Glu Gln
20 25 30Asp Ser Glu Ile Val Val Gly Lya Glu Gln Ala Arg Ala Lya Val Thr
35 40 45Gln Ser Ile Val Phe Val Thr Gly Glu Ala Ser Pro Tyr Ala Lys Ser
50 55 60Gly Gly Leu Gly Asp Val Cys Gly Ser Leu Pro Val Ala Leu Ala Ala65 70 75 80Arg Gly His Arg Val Met Val Val Met Pro Arg Tyr Leu Asn Gly Thr
85 90 95Ser Asp Lys Asn Tyr Ala Asn Ala Phe Tyr Thr Glu Lye His Ile Arg
100 105 110Ile Pro Cys Phe Gly Gly Glu His Glu Val Thr Phe Phe His Glu Tyr
115 120 125Arg Asp Ser Val Asp Trp Val Phe Val Asp His Pro Ser Tyr His Arg
130 135 140Pro Gly Asn Leu Tyr Gly Asp Lys Phe Gly Ala Phe Gly Asp Asn Gln145 150 155 160Phe Arg Tyr Thr Leu Leu Cys Tyr Ala Ala Cys Glu Ala Pro Leu Ile
165 170 175Leu Glu Leu Gly Gly Tyr Ile Tyr Gly Gln Asn Cys Met Phe Val Val
180 185 190Asn Asp Trp His Ala Ser Leu Val Pro Val Leu Leu Ala Ala Lys Tyr
195 200 205Arg Pro Tyr Gly Val Tyr Lys Asp Ser Arg Ser Ile Leu Val Ile His
210 215 220Asn Leu Ala His Gln Gly Val Glu Pro Ala Ser Thr Tyr Pro Asp Leu225 230 235 240Gly Leu Pro Pro Glu Trp Tyr Gly Ala Leu Glu Trp Val Phe Pro Glu
245 250 255Trp Ala Arg Arg His Ala Leu Asp Lys Gly Glu Ala Val Asn Phe Leu
260 265 270Lys Gly Ala Val Val Thr Ala Asp Arg Ile Val Thr Val Ser Lys Gly
275 280 285Tyr Ser Trp Glu Val Thr Thr Ala Glu Gly Gly Gln Gly Leu Asn Glu
290 295 300Leu Leu Ser Ser Arg Lys Ser Val Leu Asn Gly Ile Val Asn Gly Ile305 310 315 320Asp Ile Asn Asp Trp Asn Pro Ala Thr Asp Lys Cys Ile Pro Cys His
325 330 335Tyr Ser Val Asp Asp Leu Ser Gly Lys Ala Lys Cys Lys Gly Ala Leu
340 345 350Gln Lys Glu Leu Gly Leu Pro Ile Arg Pro Asp Val Pro Leu Ile Gly
355 360 365Phe Ile Gly Arg Leu Asp Tyr Gln Lys Gly Ile Asp Leu Ile Gln Leu
370 375 380Ile Ile Pro Asp Leu Met Arg Glu Asp Val Gln Phe Val Met Leu Gly385 390 395 400Ser Gly Asp Pro Glu Leu Glu Asp Trp Met Arg Ser Thr Glu Ser Ile
405 410 415Phe Lys Asp Lys Phe Arg Gly Trp Val Gly Phe Ser Val Pro Val Ser
420 425 430His Arg Ile Thr Ala Gly Cys Asp Ile Leu Leu Met Pro Ser Arg Phe
435 440 445Glu Pro Cys Gly Leu Asn Gln Leu Tyr Ala Met Gln Tyr Gly Thr Val
450 455 460Pro Val Val His Ala Thr Gly Gly Leu Arg Asp Thr Val Glu Asn Phe465 470 475 480Asn Pro Phe Gly Glu Asn Gly Glu Gln Gly Thr Gly Trp Ala Phe Ala
485 490 495Pro Leu Thr Thr Glu Asn Met Phe Val Asp Ile Ala Asn Cys Asn Ile
500 505 510Tyr Ile Gln Gly Thr Gln Val Leu Leu Gly Arg Ala Asn Glu Ala Arg
515 520 525His Val Lys Arg Leu His Val Gly Pro Cys Arg ★
530 535 540(2)序列22资料:
(i)序列特征:
(A)长度:30个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:线形
(ii)分子类型:其它核酸
(A)说明:/desc=“寡核苷酸”
(iii)假说:无
(xi)序列描述:序列22:GTGGATCCAT GGCGACGCCC TCGGCCGTGG(2)序列23资料:
(i)序列特征:
(A)长度:35个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:线形
(ii)分子类型:其它核酸
(A)说明:/desc=“寡核苷酸”
(ii i)假说:无
(xi)序列描述:序列23:CTGAATTCCA TATGGGGCCC CTCCCTGCTC AGCTC(2)序列24资料:
(i)序列特征:
(A)长度:36个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:线形
(ii)分子类型:其它核酸
(A)说明:/desc;“寡核苷酸”
(iii)假说:无
(xi)序列描述:序列24:CTCTGAGCTC AAGCTTGCTA CTTTCTTTCC TTAATG(2)序列25资料:
(i)序列特征:
(A)长度:29个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:线形
(ii)分子类型:其它核酸
(A)说明:/desc=“寡核苷酸”
(iii)假说:无
(xi)序列描述:序列25:GTCTCCGCGG TGGTGTCCTT GCTTCCTAG(2)序列26资料:
(i)序列特征:
(A)长度:53个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(xi)序列描述:序列26:TGCGTCGCGG AGCTGAGCAG GGAGGTCTCC GCGGTGGTGT CCTTGCTTCCTAG(2)序列27资料:
(i)序列特征:
(A)长度:8个氨基酸
(B)类型:氨基酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:肽
(xi)序列描述:序列27:Cys Val Ala Glu Leu Ser Arg Glu1 5(2)序列28资料:
(i)序列特征:
(A)长度:16个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(xi)序列描述:序列28:AGAGAGAGAG AGAGAG(2)序列29资料:
(i)序列特征:
(A)长度:36个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(xi)序列描述:序列29:AAGAAGAAGA AGAAGAAGAA GAAGAAGAAG AAGAAG(2)序列30资料:
(i)序列特征:
(A)长度:18个碱基对
(B)类型:核酸
(C)链型:双
(D)拓扑结构:不相关
(ii)分子类型:cDNA到mRNA
(iii)假说:无
(xi)序列描述:序列30:AAAAAAAAAA AAAAAAAA(2)序列31资料:
(i)序列特征:
(A)长度:11个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:其它核酸
(A)说明:/desc=“寡核苷酸”
(iii)假说:无
(xi)序列描述:序列31:AGATAATGCA G(2)序列32资料:
(i)序列特征:
(A)长度:10个碱基对
(B)类型:核酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:其它核酸
(A)说明:/desc=“寡核苷酸”
(iii)假说:无
(xi)序列描述:序列32:AACAATGGCT(2)序列33资料:
(i)序列特征:
(A)长度:56个氨基酸
(B)类型:氨基酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:肽
(iii)假说:无
(xi)序列描述:序列33:Met Ala Ser Ser Met Leu Ser Ser Ala Ala Val Ala Thr Arg Thr Asn1 5 10 15Pro Ala Gln Ala Ser Met Val Ala Pro Phe Thr Gly Leu Lys Ser Ala
20 25 30Ala Phe Pro Val Ser Arg Lys Gln Asn Leu Asp Ile Thr Ser Ile Ala
35 40 45Ser Asn Gly Gly Arg Val Gln Cys
50 55(2)序列34资料:
(i)序列特征:
(A)长度:58个氨基酸
(B)类型:氨基酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:肽
(iii)假说:无
(xi)序列描述:序列34:Met Ala Pro Thr Val Met Met Ala Ser Ser Ala Thr Ala Thr Arg Thr1 5 10 15Asn Pro Ala Gln Ala Ser Ala Val Ala Pro Phe Gln Gly Leu Lys Ser
20 25 30Thr Ala Ser Leu Pro Val Ala Arg Arg Ser Ser Arg Ser Leu Gly Asn
35 40 45Val Ala Ser Asn Gly Gly Arg Ile Arg Cys
50 55(2)序列35资料:
(i)序列特征:
(A)长度:58个氨基酸
(B)类型:氨基酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:肽
(iii)假说:无
(xi)序列描述:序列35:Met Ala Gln Ile Leu Ala Pro Ser Thr Gln Trp Gln Met Arg Ile Thr1 5 10 15Lys Thr Ser Pro Cys Ala Thr Pro Ile Thr Ser Lys Met Trp Ser Ser
20 25 30Leu Val Met Lys Gln Thr Lys Lys Val Ala His Ser Ala Lys Phe Arg
35 40 45Val Met Ala Val Asn Ser Glu Asn Gly Thr
50 55(2)序列36资料:
(i)序列特征:
(A)长度:74个氨基酸
(B)类型:氨基酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:肽
(iii)假说:无
(xi)序列描述:序列36:Met Ala Ala Leu Ala Thr Ser Gln Leu Val Ala Thr Arg Ala Gly His1 5 10 15Gly Val Pro Asp Ala Ser Thr Phe Arg Arg Gly Ala Ala Gln Gly Leu
20 25 30Arg Gly Ala Arg Ala Ser Ala Ala Ala Asp Thr Leu Ser Met Arg Thr
35 40 45Ser Ala Arg Ala Ala Pro Arg His Gln Gln Gln Ala Arg Arg Gly Gly
50 55 60Arg Phe Pro Phe Pro Ser Leu Val Val Cys65 70(2)序列37资料:
(i)序列特征:
(A)长度:39个氨基酸
(B)类型:氨基酸
(C)链型:单
(D)拓扑结构:不相关
(ii)分子类型:肽
(iii)假说:无
(xi)序列描述:序列37:Met Ala Thr Pro Ser Ala Val Gly Ala Ala Cys Leu Leu Leu Ala Arg1 5 10 15Xaa Ala Trp Pro Ala Ala Val Gly Asp Arg Ala Arg Pro Arg Arg Leu
20 25 30Gln Arg Val Leu Arg Arg Arg
35
Claims (20)
1.一种杂合多肽,它包括:
(a)一个淀粉被囊化区;
(b)一个与所述淀粉被囊化区融合的有效负荷多肽。
2.如权利要求1所述的杂合多肽,其中,所述有效负荷多肽由不超过三种以上的不同类型的氨基酸组成,所述氨基酸选自下列一组:Ala,Arg,Asn,Asp,Cys,Gln,Glu,Gly,His,Ile,Leu,Lys,Met,Phe,Pro,Ser,Thr,Trp,Tyr,和Val。
3.如权利要求1所述的杂合多肽,其中,所述有效负荷多肽是一种生物活性多肽。
4.如权利要求3所述的杂合多肽,其中,所述有效负荷多肽选自下列一组:激素,生长因子,抗体,肽,多肽,酶免疫球蛋白,染料及其生物活性片段。
5.如权利要求1所述的杂合多肽,其中,所述淀粉被囊化区是一种酶的淀粉被囊化区,所述酶选自下列一组:可溶性淀粉合成酶I、可溶性淀粉合成酶II、可溶性淀粉合成酶III、颗粒结合淀粉合成酶、分支酶I、分支酶IIa、分支酶IIBb和葡糖淀粉酶多肽。
6.如权利要求1所述的杂合多肽,它包括一个位于所述淀粉被囊化区和有效负荷多肽之间的裂解位点。
7.一种编码权利要求1所述杂合多肽的核酸分子。
8.如权利要求7的重组分子,它是一种DNA分子,该分子包括适于在细菌宿主中表达所述淀粉被囊化区和所述有效负荷多肽的控制序列。
9.如权利要求7的重组分子,它是一种DNA分子,该分子包括适于在植物宿主中表达所述淀粉被囊化区和所述有效负荷多肽的控制序列。
10.如权利要求9的重组分子,其中,所述控制序列适于在单子叶植物中表达所述淀粉被囊化区和所述有效负荷多肽。
11.如权利要求9的重组分子,其中,所述控制序列适于在双子叶植物中表达所述淀粉被囊化区和所述有效负荷多肽。
12.如权利要求9的重组分子,其中,所述控制序列适于在动物宿主中表达所述淀粉被囊化区和所述有效负荷多肽。
13.一种含有权利要求7所述重组分子的表达载体。
14.一种被转化成含有权利要求7所述重组分子的细胞,该细胞能表达所述DNA分子。
15.如权利要求14的细胞,它是一种植物细胞。
16.一种由权利要求15所述细胞再生的植物。
17.一种由权利要求16所述植物所结的种子,该种子能表达所述重组分子。
18.一种由权利要求14所述细胞产生的改性淀粉,它包括所述有效负荷多肽。
19.一种在动物的消化系统的特定部位中对有效负荷多肽进行定向消化的方法,该方法包括给动物喂饲权利要求18所述的改性淀粉,该淀粉包括存在于淀粉基质中的有效负荷多肽,该淀粉经过选择,以便能在其消化道的特定部位被消化。
20.一种用权利要求1所述杂合多肽生产一种纯的有效负荷多肽的方法,该方法包括:
(a)用编码所述杂合多肽的DNA转化一种宿主生物;
(b)让所述杂合多肽在所述宿主中表达;
(c)从所述宿主中分离所述杂合多肽;
(d)从所述杂合多肽中纯化所述有效负荷多肽。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US2685596P | 1996-09-30 | 1996-09-30 | |
US60/026,855 | 1996-09-30 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1239514A true CN1239514A (zh) | 1999-12-22 |
CN1195861C CN1195861C (zh) | 2005-04-06 |
Family
ID=21834174
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNB971802327A Expired - Fee Related CN1195861C (zh) | 1996-09-30 | 1997-09-30 | 将多肽被囊化于淀粉基质中 |
Country Status (14)
Country | Link |
---|---|
US (2) | US6107060A (zh) |
EP (1) | EP0935665B1 (zh) |
JP (1) | JP2001505412A (zh) |
KR (1) | KR20000048782A (zh) |
CN (1) | CN1195861C (zh) |
AT (1) | ATE389725T1 (zh) |
AU (1) | AU730427B2 (zh) |
BR (1) | BR9713242A (zh) |
CA (1) | CA2265514A1 (zh) |
DE (1) | DE69738587T2 (zh) |
IL (1) | IL129158A0 (zh) |
NZ (1) | NZ334637A (zh) |
RU (1) | RU2268301C9 (zh) |
WO (1) | WO1998014601A1 (zh) |
Families Citing this family (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6960457B1 (en) * | 1997-09-04 | 2005-11-01 | Stanford University | Reversible immobilization of arginine-tagged moieties on a silicate surface |
DE19820607A1 (de) * | 1998-05-08 | 1999-11-11 | Hoechst Schering Agrevo Gmbh | Nucleinsäuremoleküle codierend Enzyme aus Weizen, die an der Stärkesynthese beteiligt sind |
US6392120B1 (en) | 1998-07-28 | 2002-05-21 | E. I. Du Pont De Nemours And Company | Modification of starch biosynthetic enzyme gene expression to produce starches in grain crops |
AUPQ005299A0 (en) * | 1999-04-29 | 1999-05-27 | Commonwealth Scientific And Industrial Research Organisation | Novel genes encoding wheat starch synthases and uses therefor |
FR2793806B1 (fr) * | 1999-05-21 | 2003-04-25 | Centre Nat Rech Scient | Grains d'amidon contenant un polypeptide recombinant d'interet, leur procede d'obtention, et leurs utilisations |
US7135619B1 (en) | 1999-06-11 | 2006-11-14 | Wageningen Universiteit | Expression in plants of starch binding domains and/or of protein-fusions containing starch binding domains |
US7041484B1 (en) | 1999-10-29 | 2006-05-09 | National Research Council Of Canada | Starch branching enzymes |
AU1122701A (en) * | 1999-10-29 | 2001-05-14 | National Research Council Of Canada | Starch branching enzymes |
WO2002079410A2 (en) * | 2001-03-30 | 2002-10-10 | Basf Plant Science Gmbh | Glucan chain length domains |
EP2216405A1 (en) | 2002-05-03 | 2010-08-11 | Monsanto Technology LLC | Speed specific USP promoters for expressing genes in plants |
EP1648996B1 (en) | 2003-06-25 | 2012-03-14 | Novozymes A/S | Enzymes for starch processing |
ATE510925T1 (de) | 2003-06-25 | 2011-06-15 | Novozymes As | Stärkehydrolyseverfahren |
ATE517994T1 (de) | 2003-06-30 | 2011-08-15 | Commw Scient Ind Res Org | Weizen mit veränderter verzweigungsenzymaktivität sowie daraus erhaltene stärke und stärkehaltige produkte |
KR101274849B1 (ko) * | 2003-10-27 | 2013-06-13 | 코몬웰스 싸이언티픽 엔드 인더스트리얼 리서치 오가니제이션 | 아밀로스 비율이 증가된 전분을 갖는 쌀 및 이의 생성물 |
US9756798B2 (en) | 2004-11-19 | 2017-09-12 | Patti D. Rubin | Burrow filling compressed growing medium |
US20060107589A1 (en) | 2004-11-19 | 2006-05-25 | Rubin Patti D | Compressed growing medium |
DK1831385T3 (en) | 2004-12-22 | 2015-07-13 | Novozymes North America Inc | Enzymes for starch processing |
US8841091B2 (en) | 2004-12-22 | 2014-09-23 | Novozymes Als | Enzymes for starch processing |
FR2911608B1 (fr) * | 2007-01-23 | 2009-04-03 | Centre Nat Rech Scient | Nouvelles compositions vaccinales anti paludique et ses utilisations. |
US20090113791A1 (en) | 2007-10-29 | 2009-05-07 | Oms Investments, Inc. | Compressed Coconut Coir Pith Granules and Methods for the Production and use Thereof |
ES2377617B1 (es) * | 2010-08-31 | 2012-11-23 | Iden Biotechnology, S.L. | Procedimiento para la producción y purificación de proteínas recombinantes en plantas. |
GB201311272D0 (en) | 2013-06-25 | 2013-08-14 | Ucl Business Plc | Anti-microbial agents and uses thereof |
KR101949009B1 (ko) * | 2016-02-18 | 2019-02-18 | 경희대학교 산학협력단 | 융합단백질-다당 복합체의 제조방법 및 이의 용도 |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4859377A (en) * | 1987-07-10 | 1989-08-22 | The United States Of America, As Represented By The Secretary Of Agriculture | Starch encapsulation of entomopathogens |
US5137819A (en) * | 1988-07-08 | 1992-08-11 | University Of British Columbia | Cellulose binding fusion proteins for immobilization and purification of polypeptides |
US5302523A (en) * | 1989-06-21 | 1994-04-12 | Zeneca Limited | Transformation of plant cells |
US5349123A (en) * | 1990-12-21 | 1994-09-20 | Calgene, Inc. | Glycogen biosynthetic enzymes in plants |
US5643756A (en) * | 1992-08-28 | 1997-07-01 | The Public Health Research Institute Of The City Of New York, Inc. | Fusion glycoproteins |
GB9307408D0 (en) * | 1993-04-08 | 1993-06-02 | Danisco | Transgenic plants |
US5512459A (en) * | 1993-07-20 | 1996-04-30 | Bionebraska, Inc. | Enzymatic method for modification or recombinant polypeptides |
US5648244A (en) * | 1993-09-27 | 1997-07-15 | President And Fellows Of Harvard College | Production, purification, cleavage and use of fusion peptides |
US5635599A (en) * | 1994-04-08 | 1997-06-03 | The United States Of America As Represented By The Department Of Health And Human Services | Fusion proteins comprising circularly permuted ligands |
CA2624375C (en) * | 1996-05-29 | 2010-07-27 | Bayer Cropscience Gmbh | Nucleic acid molecules encoding enzymes from wheat which are involved in starch synthesis |
-
1997
- 1997-09-30 NZ NZ334637A patent/NZ334637A/en unknown
- 1997-09-30 US US08/941,445 patent/US6107060A/en not_active Expired - Lifetime
- 1997-09-30 RU RU99108730/13A patent/RU2268301C9/ru not_active IP Right Cessation
- 1997-09-30 KR KR1019990702770A patent/KR20000048782A/ko not_active Application Discontinuation
- 1997-09-30 BR BR9713242-0A patent/BR9713242A/pt not_active Application Discontinuation
- 1997-09-30 DE DE69738587T patent/DE69738587T2/de not_active Expired - Lifetime
- 1997-09-30 CN CNB971802327A patent/CN1195861C/zh not_active Expired - Fee Related
- 1997-09-30 WO PCT/US1997/017555 patent/WO1998014601A1/en not_active Application Discontinuation
- 1997-09-30 IL IL12915897A patent/IL129158A0/xx unknown
- 1997-09-30 JP JP51677798A patent/JP2001505412A/ja not_active Ceased
- 1997-09-30 CA CA002265514A patent/CA2265514A1/en not_active Abandoned
- 1997-09-30 AU AU48030/97A patent/AU730427B2/en not_active Ceased
- 1997-09-30 AT AT97910730T patent/ATE389725T1/de active
- 1997-09-30 EP EP97910730A patent/EP0935665B1/en not_active Expired - Lifetime
-
2003
- 2003-07-28 US US10/628,525 patent/US7141659B2/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
KR20000048782A (ko) | 2000-07-25 |
DE69738587T2 (de) | 2009-04-30 |
WO1998014601A1 (en) | 1998-04-09 |
JP2001505412A (ja) | 2001-04-24 |
CN1195861C (zh) | 2005-04-06 |
RU2268301C9 (ru) | 2006-07-20 |
US7141659B2 (en) | 2006-11-28 |
DE69738587D1 (de) | 2008-04-30 |
RU2268301C2 (ru) | 2006-01-20 |
CA2265514A1 (en) | 1998-04-09 |
EP0935665B1 (en) | 2008-03-19 |
NZ334637A (en) | 2001-02-23 |
US20040185114A1 (en) | 2004-09-23 |
AU730427B2 (en) | 2001-03-08 |
BR9713242A (pt) | 2000-01-18 |
IL129158A0 (en) | 2000-02-17 |
AU4803097A (en) | 1998-04-24 |
EP0935665A1 (en) | 1999-08-18 |
US6107060A (en) | 2000-08-22 |
ATE389725T1 (de) | 2008-04-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1195861C (zh) | 将多肽被囊化于淀粉基质中 | |
CN1163612C (zh) | 编码蔗糖依赖性蔗糖果糖基转移酶的核酸分子及生产短链果糖基多聚物的方法 | |
CN1183256C (zh) | 在基因工程植物的贮藏器官中增加类胡萝卜素的积累 | |
CN1145698C (zh) | 合成杀虫的晶状蛋白质基因 | |
CN1257978C (zh) | 编码小麦中参与淀粉合成的酶的核酸分子 | |
CN1285875A (zh) | 突变的羟基苯丙酮酸双氧化酶、基dna序列和含该基因且耐除草剂的植物的分离 | |
CN1402789A (zh) | 来自小麦的核酸分子、转基因植物细胞和植物及其在生产改性淀粉中的应用 | |
CN1777677A (zh) | 增强植物中直链淀粉产量 | |
CN1930296A (zh) | 具有增加的淀粉磷酸化酶活性的植物 | |
CN1216583A (zh) | 编码类黄酮途径酶的基因序列及其用途 | |
CN101065491A (zh) | 生产胶原的植物及其生成和使用方法 | |
CN1212725A (zh) | 来源于植物原卟啉原氧化酶基因的启动子 | |
CN1398300A (zh) | 新延伸酶基因以及制备多不饱和脂肪酸的方法 | |
CN1246461C (zh) | 修饰植物中次级代谢化合物水平的方法和组合物 | |
CN1054170A (zh) | 可繁殖的转基因谷类植物 | |
CN1930301A (zh) | 鉴定具有淀粉磷酸化酶促活性的蛋白质的方法 | |
CN1236394A (zh) | 除草剂抗性植物 | |
CN1198777A (zh) | 有关谷氨酸脱氢酶α-和β-亚基的新多肽和多核苷酸及用法 | |
CN1487998A (zh) | 编码乙酰乳酸合酶基因的基因 | |
CN101076594A (zh) | 参与糖和脂质代谢的蛋白质的拟南芥编码基因及使用方法 | |
CN1372599A (zh) | 植物甾醇酰基转移酰 | |
CN1468311A (zh) | 异麦芽寡糖合酶 | |
CN1852975A (zh) | 转基因的高色氨酸植物 | |
CN1668754A (zh) | 胡萝卜素合酶基因及其用途 | |
CN1127016A (zh) | 提高贮存马铃薯质量的方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20050406 Termination date: 20140930 |
|
EXPY | Termination of patent right or utility model |