CN101631871B - L-氨基酸的生产方法 - Google Patents
L-氨基酸的生产方法 Download PDFInfo
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Abstract
在含有甘油作为碳源的培养基中培养具有L-氨基酸生产能力、且经过修饰使得甘油脱氢酶和二羟基丙酮激酶的活性增强的属于肠杆菌科的微生物,来生产L-氨基酸。
Description
技术领域
本发明涉及发酵技术,更详细地说,涉及利用微生物发酵的L-氨基酸的生产。
背景技术
L-氨基酸是通过使用属于短杆菌属(Brevibacterium)、棒状杆菌属(Corynebacterium)、埃希氏菌属(Escherichia)等的微生物的发酵方法来进行工业生产的。在这些生产方法中,使用从自然界分离的菌株或该菌株的人工突变株,并且还使用通过重组DNA技术进行修饰从而增加了碱性L-氨基酸生物合成酶的活性的微生物等。(专利文献1~9)
通常,在使用微生物进行氨基酸生产时,在基质中使用糖类作为主成分,但甘油也可以与糖类一样用作基质。(专利文献10、11)
已知大肠杆菌(Escherichia coli)中存在多个甘油代谢相关基因。然而,缺损甘油激酶的编码基因glpK、或甘油3磷酸脱氢酶的编码基因glpD中的任一个的突变株在以甘油为单一碳源的培养基中不能生长,由此可知:甘油激酶和甘油-3-磷酸脱氢酶担负着大肠杆菌的主要甘油同化途径。(非专利文献1)
大肠杆菌的甘油脱氢酶也是甘油代谢相关的酶之一,已知在使用缺损了glpR(glpK、glpD、glp调节子的阻遏物)的3个基因的突变株进行的筛选中,它可使在以甘油为单一碳源的培养基中的致死性得以回复。(非专利文献2)
目前为止,如上述,经由甘油-3-磷酸的甘油激酶和甘油-3-磷酸脱氢酶被认为是属于肠杆菌科的微生物的主要同化途径,而经由二羟基丙酮的甘油同化途径被认为是属于肠杆菌科的微生物的甘油同化的非必要途径。
专利文献1EP0643135B
专利文献2EP0733712B
专利文献3EP1477565A
专利文献4EP0796912A
专利文献5EP0837134A
专利文献6WO01/53459
专利文献7EP1170376A
专利文献8WO2005/010175
专利文献9WO96/17930
专利文献10EP1715055A
专利文献11EP1715056A
非专利文献1J.Bacteriol.23(2006)8259-8271
非专利文献2J.Bacteriol.131(1977)1026-1028
发明内容
发明所要解决的问题
本发明的问题是提供一种与传统方法相比进一步有所改良的、通过使用含有甘油的基质的发酵方法进行的L-氨基酸生产方法。
解决问题的方法
本发明人等为解决上述问题进行了深入研究,结果发现:虽然单独增强甘油脱氢酶和二羟基丙酮激酶这两种经由二羟基丙酮的甘油同化途径中的酶没有效果,但通过同时增强甘油脱氢酶和二羟基丙酮激酶,可以大幅提高从甘油生产L-氨基酸的能力,从而完成了本发明。
即,本发明如下所述。
(1)一种生产L-氨基酸的方法,包括在含有甘油作为碳源的培养基中培养具有L-氨基酸生产能力且经过修饰而增强了甘油脱氢酶和二羟基丙酮激酶的活性的属于肠杆菌科的微生物,从而在该培养基中或菌体内生成和蓄积L-氨基酸,并且从该培养基或菌体收集L-氨基酸。
(2)(1)所述的方法,其中,所述酶的活性是通过提高编码所述甘油脱氢酶和二羟基丙酮激酶的基因的拷贝数或者通过修饰所述基因的表达调节序列来增强的。
(3)(1)~(2)所述的方法,其特征在于,所述二羟基丙酮激酶以ATP为磷酸供体。
(4)(1)~(3)中任一项所述的方法,其中,所述微生物还经过修饰而增强了甘油的摄入活性。
(5)(1)~(4)中所述的方法,其中,所述微生物还经过修饰而增强了选自由丙糖磷酸异构酶、果糖二磷酸醛缩酶、果糖1,6-二磷酸酶和果糖-6-磷酸醛缩酶组成的群组中的1种以上的酶的酶活性。
(6)(1)~(5)中所述的方法,其中,所述微生物还经过修饰而降低了甘油激酶和/或膜结合型甘油-3-磷酸脱氢酶的酶活性。
(7)(1)~(6)中所述的方法,其中,所述属于肠杆菌的微生物为埃希氏菌属细菌或泛菌属(Pantoea)细菌。
(8)(1)~(7)中所述的方法,其中,所述L-氨基酸为选自由L-谷氨酸、L-赖氨酸、L-亮氨酸、L-异亮氨酸、L-缬氨酸、L-色氨酸、L-苯丙氨酸、L-酪氨酸、L-苏氨酸、L-甲硫氨酸、L-半胱氨酸、L-精氨酸、L-丝氨酸、L-脯氨酸、L-天冬氨酸、L-天冬酰胺、L-谷氨酰胺和L-组氨酸组成的群组中的1种或2种以上的L-氨基酸。
实施本发明的最佳模式
以下,对本发明进行详细说明。
<1>本发明的微生物
本发明的微生物是具有L-氨基酸生产能力,且经过修饰而增强了甘油脱氢酶和二羟基丙酮激酶的活性增强的属于肠杆菌科的微生物。这里,L-氨基酸生产能力是指:当在培养基中培养本发明的微生物时,在培养基中或菌体内生成并蓄积L-氨基酸的能力。而且,本发明的微生物可以是具有多种L-氨基酸的生产能力的微生物。作为具有L-氨基酸生产能力的微生物,可以是本身具有L-氨基酸生产能力的微生物,也可以是采用突变方法或重组DNA技术对如下所述的微生物进行修饰而具有了L-氨基酸生产能力的微生物。
对于L-氨基酸的种类没有特殊限制,可以列举:L-赖氨酸、L-鸟氨酸、L-精氨酸、L-组氨酸、L-瓜氨酸等碱性氨基酸,L-异亮氨酸、L-丙氨酸、L-缬氨酸、L-亮氨酸、L-甘氨酸等脂肪族氨基酸,L-苏氨酸、L-丝氨酸等作为羟基单氨基羧酸的氨基酸,L-脯氨酸等环式氨基酸,L-苯丙氨酸、L-酪氨酸、L-色氨酸等芳香族氨基酸,L-半胱氨酸、L-胱氨酸、L-甲硫氨酸等含硫氨基酸,L-谷氨酸、L-天冬氨酸等酸性氨基酸,L-谷氨酰胺、L-天冬酰胺等侧链具有酰胺基的氨基酸。本发明的微生物可以是具有2种以上氨基酸的生产能力的微生物。
作为本发明的属于肠杆菌科的微生物,可以使用以埃希氏菌属细菌、泛菌属细菌为代表的属于肠杆菌科的微生物。作为其它的属于肠杆菌科的微生物,可以列举:肠杆菌属(Enterobacter)、克雷伯氏菌属(Klebsiella)、沙雷氏菌属(Serratia)、欧文氏菌属(Erwinia)、沙门氏菌属(Salmonella)、摩根氏菌属(Morganella)等属于γ-变形细菌的属于肠杆菌科的微生物。
在本发明中,“甘油脱氢酶”是指:以NAD为辅酶,可逆地催化将甘油转变为二羟基丙酮的以下氧化反应的酶(EC:1.1.1.6)。
甘油(glycerol)+NAD=二羟基丙酮(dihydroxyacetone)+NADH+H+
在本发明中,“经过修饰而升高了甘油脱氢酶的活性”对应于下列情形:相对于野生株或非修饰株,平均每个细胞的甘油脱氢酶分子数增加,或者甘油脱氢酶的平均每个分子的活性提高。此外,当在野生株中不能检出的酶活性提高到能够检出的程度时,这种情况也包含在所述的“活性升高”中。这里,在本发明中,甘油脱氢酶的活性只要能够检出即可,优选经过修饰从而具有0.05U/mg以上、优选0.25U/mg以上、更优选0.5U/mg以上的酶活性。这里,作为属于肠杆菌科的微生物中的作为对照的野生株,可以列举大肠杆菌MG1655株(ATCC No.47076)、和W3110株(ATCC No.27325)、Pantoea ananatis AJ13335株(FERM BP-6615)等。甘油脱氢酶活性可以参考Ansis,R.E等的方法来测定((1953)J.Biol.Chem.2-3,153-159)。
在本发明中,“二羟基丙酮激酶”是指:可逆地催化将二羟基丙酮转变为磷酸二羟基丙酮的以下反应的酶,该酶中有的以ATP为磷酸供体(EC2.7.1.29),有的以PEP为磷酸供体(EC 2.7.1.29)(Cell.Mol.Life Sci.63(2006)890-900,Biochemistry.43(2004)13037-13045)。
ATP+二羟基丙酮(dihydroxyacetone)=ADP+磷酸二羟基丙酮(dihydroxyacetone phosphate)(EC 2.7.1.29)
磷酸烯醇丙酮酸(phosphoenolpyruvate)+二羟基丙酮(dihydroxyacetone)=丙酮酸(pyruvate)+磷酸二羟基丙酮(dihydroxyacetone phosphate)(EC2.7.1.29)
在本发明中,特别优选使用以ATP为磷酸供体的二羟基丙酮激酶。
“经过修饰而升高了二羟基丙酮激酶活性”对应于这样的情况:与野生株或非修饰株相比,每个细胞平均的二羟基丙酮激酶分子数增加,或者二羟基丙酮激酶的平均每个分子的活性提高。此外,当在野生株中不能检出的酶活性提高到能够检出的程度时,这种情况也包含在所述的“活性升高”中。优选经过修饰使得二羟基丙酮激酶的活性与野生株或非修饰株相比,平均每个菌体提高150%以上、优选200%以上,更希望平均每个菌体提高300%以上。这里,作为对照的属于肠杆菌科的微生物的野生株包括例如大肠杆菌MG1655株(ATCC No.47076)和W3110株(ATCC No.27325)、Pantoeaananatis AJ13335株(FERM BP-6615)等。二羟基丙酮激酶活性可以参考Johnson EA等的方法来测定(J Bacteriol.1984 Oct;160(1):55-60)。
编码甘油脱氢酶的基因包括例如gldA基因,这其中希望使用属于肠杆菌科的微生物来源的gldA基因。作为属于肠杆菌科的微生物,可以列举大肠杆菌。这里,作为大肠杆菌的基因,可以列举例如SEQ ID NO:1的gldA基因(GenBank登录号NC_000913的碱基序号4135955..4137058的互补链)。
而且,编码甘油脱氢酶的基因的同源物可以是基于与上述示例的基因的同源性,从埃希氏菌属、肠杆菌属、克雷伯氏菌属、沙雷氏菌属、欧文氏菌属、耶尔森菌属(Yersinia)、志贺氏菌属(Shigella)、沙门氏菌属、弧菌属(vibrio)、气单胞菌属(Aeromonas)、芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)、乳杆菌属(Lactobacillus)、肠球菌属(Enterococcus)、梭菌属(Clostridium)、假单胞菌属(Pseudomonas)、土壤杆菌属(Agrobacterium)、柠檬酸杆菌属(Citrobacter)、棒状杆菌属细菌等克隆得到的基因。表1示出了与大肠杆菌gldA基因具有高同源性、可以在本发明中作为编码甘油脱氢酶的基因利用的基因的例子。
[表1]
与大肠杆菌gldA基因的同源性高的、编码甘油脱氢酶的基因
基因 | 微生物 | 说明 | Genbank登录号 | 序列表序列号 |
gldA | 痢疾志贺氏菌Sd197 | 甘油脱氧酶,(NAD) | YP_405216.1GI:82778867 | 74、75 |
gldA | 鼠伤寒沙门氏菌LT2 | 与大肠杆菌甘油脱氢酶类似,(NAD) | AE008892.1GI:16422675 | 76、77 |
gldA | 恶臭假单胞菌 | 甘油脱氧酶 | AF148496.1GI:6552505 | 78、79 |
gldA | 凝结芽孢杆菌 | 甘油脱氢酶和相关酶 | ZP_01697292.1GI:124522908 | 80、81 |
氨基酸序列和碱基序列的同源性(同一性等)可以使用例如Karlin和Altschul的算法BLAST(Pro.Natl.Acad.Sci.USA,90,5873(1993))或Pearson的FASTA(Methods Enzymol.,183,63(1990))来确定。基于该算法BLAST,开发了称为BLASTN、BLASTX的程序(参照http://www.ncbi.nlm.nih.gov)。
本发明的编码二羟基丙酮激酶的基因可以利用命名为dhaKLM基因、dak1基因、dhaK基因、dhbK基因的基因。这里,作为以PEP为磷酸供体的基因,可以列举大肠杆菌来源的基因,例如可以列举SEQ ID NO:34的dhaK基因(GenBank登录号NC_000913的碱基序号1248991..1250061的互补链)、SEQ ID NO:36的dhaL基因(GenBank登录号NC_000913的碱基序号1248348..1248980的互补链)、SEQ ID NO:38的dhaM基因(GenBank登录号NC_000913的碱基序号1246919..1248337的互补链)。
在本发明中,更优选使用编码以ATP为磷酸供体的二羟基丙酮激酶的基因,作为以ATP为磷酸供体的二羟基丙酮激酶的基因,优选使用酵母来源的dak1基因、土壤杆菌属来源的dhbK基因、柠檬酸杆菌属来源的dhaK基因。作为酵母来源的dak1基因,可以列举酿酒酵母(saccharomycescerevisiae)来源的SEQ ID NO:3的dak1基因(GenBank Accession NoNP_013641.1GI:6323570),作为土壤杆菌的dhbK基因,可以列举根瘤土壤杆菌(Agrobacterium tumefaciens)来源的SEQ ID NO:5的dhbK基因(GenBank登录号NP_357070.1GI:15891398),作为柠檬酸杆菌基因的dhaK基因,可以列举弗氏柠檬酸杆菌(Citrobacter freundii)来源的SEQ IDNO:7的dhaK基因(GenBank登录号U09771)。
而且,编码二羟基丙酮激酶的基因的同源物,可以是基于与以上示例的基因的同源性,从埃希氏菌属、肠杆菌属、克雷伯氏菌属、沙雷氏菌属、欧文氏菌属、耶尔森氏菌属、志贺氏菌属、沙门氏菌属、弧菌属、气单胞菌属、芽孢杆菌属、葡萄球菌属、乳杆菌属、肠球菌属、梭菌属、土壤杆菌属、柠檬酸杆菌属、分枝杆菌属(Mycobacterium)等的细菌、酵母属、裂殖酵母属(Schizosaccharomyces)、毕赤酵母属(Pichia)等酵母等中克隆得到的基因。
特别是以ATP为磷酸供体的二羟基丙酮激酶,可以使用以下序列。表2示出了与酿酒酵母来源的dak1基因高同源性的编码二羟基丙酮激酶基因,表3示出了与根瘤土壤杆菌(Agrobacterium tumefaciens)来源的dhbK基因同源性高的二羟基丙酮激酶基因,表4示出了与弗氏柠檬酸杆菌来源的dhaK基因同源性高的二羟基丙酮激酶基因。
[表2]
与酿酒酵母dak1基因的同源性高的、编码二羟基丙酮激酶的基因
基因 | 微生物 | 说明 | Genbank登录号 | 序列表序列号 |
T43702 | 栗酒裂殖酵母 | 二羟基丙酮激酶 | gi|3493578|gb|AAC78808.1| | 40、41 |
AAC27705 | Pichia angusta | 二羟基丙酮激酶 | gi|3171001|gb|AAC27705.1| | 42、43 |
AAC39490.1 | 巴斯德毕赤酵母 | 二羟基丙酮激酶 | gi|3287486|gb|AAC39490.1| | 44、45 |
CAG88710.1 | 汉森德巴利酵母CBS767 | 二羟基丙酮激酶 | gi|49656075|emb|CAG88710.1| | 46、47 |
[表3]
与根瘤土壤杆菌dhbK基因的同源性高的、编码二羟基丙酮激酶的基因
基因 | 微生物 | 说明 | Genbank登录号 | 序列表序列号 |
ABF89849.1 | 黄色粘球菌DK 1622 | 二羟基丙酮激酶家族蛋白 | gi|108464664|gb|ABF89849.1| | 58、59 |
ABB06761.1 | 伯克霍尔德氏菌383 | 甘油酮激酶 | gi|77965380|gb|ABB06761.1|Glycerone kinase[Burkholderia sp.383] | 60、61 |
ABC38950.1 | 泰国伯克霍尔德氏菌E264 | 二羟基丙酮激酶 | gi|83654887|gb|ABC38950.1| | 62、63 |
EAV65448.1 | 多食伯克霍尔德氏菌ATCC 17616 | 甘油酮激酶 | gi|118658702|gb|EAV65448.1| | 64、65 |
[表4]
与弗氏柠檬酸杆菌dhaK基因同源性高的、编码二羟基丙酮激酶的基因
基因 | 微生物 | 说明 | Genbank登录号 | 序列表序列号 |
AAX12907.1 | 蟑螂埃希氏菌 | 二羟基丙酮激酶蛋白 | gi|60099603|gb|AAX12907.1| | 48、49 |
EAV82971.1 | 肠杆菌638 | 二羟基丙酮激酶蛋白 | gi|118676428|gb|EAV82971.1| | 50、51 |
EAS39398.1 | 嗜冷单胞菌CNPT3 | 二羟基丙酮激酶蛋白 | gi|90311294|gb|EAS39398.1| | 52、53 |
EAV42339.1 | Stappia aggregataIAM 12614 | 二羟基丙酮激酶蛋白 | gi|118435694|gb|EAV42339.1| | 54、55 |
CAK08390.1 | 豌豆根瘤菌蚕豆生物变种3841 | 推定的二羟基丙酮激酶 | gi|115257295|emb|CAK08390.1| | 56、57 |
上述基因的同源物是指:其它微生物来源的、或者天然或人工的突变型基因,显示与选自上述基因群的基因高度的结构类似性,在导入宿主或使其在宿主中扩增时具有提高甘油脱氢酶和二羟基丙酮激酶活性的功能。甘油脱氢酶和二羟基丙酮激酶基因的同源物是指:编码分别相对于SEQ IDNO:2、4、6或8、表1~4所述的序列的氨基酸序列全长具有80%以上、优选90%以上、更优选95%、特别优选98%以上的同源性,且具有作为甘油脱氢酶或二羟基丙酮激酶的功能的蛋白的基因。而且,是否具有甘油脱氢酶或二羟基丙酮激酶活性,可以通过使这些基因在宿主细胞中表达,采用上述酶活性测定方法考察与非修饰株相比酶活性上升与否来确认。此外,是否为同源物可以通过制备对应野生型基因的破坏株,考察将其导入破坏株时是否与野生型基因的功能互补来确定,例如通过考察因基因破坏而降低的酶活性是否恢复来确定。
此外,就本发明中使用的编码甘油脱氢酶和二羟基丙酮激酶的基因而言,不仅限于野生型基因,只要所编码的甘油脱氢酶和二羟基丙酮激酶的功能不受到破坏即可,可以是编码下述蛋白的突变体或人工修饰体,所述蛋白具有在SEQ ID NO:2,4,6或8的氨基酸序列、表1~4所述的氨基酸序列中包含1个或多个位置上的1个或数个氨基酸取代、缺失、插入或添加的序列。这里,“1个或数个”因氨基酸残基的种类或在蛋白立体结构中的位置而异,但具体来说是1~20个、优选1~10个、更优选1~5个、更优选1~3个。上述取代优选是保守取代。关于保守突变,当取代位点是芳香族氨基酸时为Phe、Trp和Tyr之间、当取代位点是疏水氨基酸时为Leu、Ile和Val之间、是极性氨基酸时为Gln和Asn之间、当是碱性氨基酸时为Lys、Arg和His之间、当是酸性氨基酸时为Asp和Glu之间、当是具有羟基的氨基酸时为Ser和Thr之间的相互取代的突变。作为保守取代可列举:用Ser或Thr取代Ala;用Gln、His或Lys取代Arg;用Glu、Gln、Lys、His或Asp取代Asn;用Asn、Glu或Gln取代Asp;用Ser或Ala取代Cys;用Asn、Glu、Lys、His、Asp或Arg取代Gln;用Gly、Asn、Gln、Lys或Asp取代Glu;用Pro取代Gly;用Asn、Lys、Gln、Arg或Tyr取代His;用Leu、Met、Val或Phe取代Ile;用Ile、Met、Val或Phe取代Leu;用Asn、Glu、Gln、His或Arg取代Lys;用Ile、Leu、Val或Phe取代Met;用Trp、Tyr、Met、Ile或Leu取代Phe;用Thr或Ala取代Ser;用Ser或Ala取代Thr;用Phe或Tyr取代Trp;用His、Phe或Trp取代Tyr;和用Met、Ile或Leu取代Val。此外,这样的氨基酸的取代、缺失、插入、添加或倒位等还包括因基于携带kdp操纵子的微生物的个体差异、种间差异等情形而天然发生的突变(mutant或variant)而产生的那些取代、缺失、插入、添加或倒位。
此外,编码甘油脱氢酶和二羟基丙酮激酶的基因还包括下述DNA,所述DNA与分别由SEQ ID NO:2、4、6或8所示的碱基序列、表1~4所述的碱基序列的互补碱基序列,或可从上述序列制备的探针在严格条件下杂交,并且编码具有作为甘油脱氢酶和二羟基丙酮激酶的功能的蛋白。这里,“严格条件”是指形成所谓特异性杂交体而不形成非特异性杂交体的条件。将该条件明确地数值化是困难的,举一实例,有这样的条件:在该条件下,具有高同源性的DNA,例如具有80%以上,优选90%以上,更优选95%以上,特别优选具有98%以上同源性的DNA相互杂交,而同源性较之为低的DNA则不互相杂交;或者是这样的条件:在与常规Southern杂交的洗涤条件即60℃,1×SSC、0.1%SDS,优选0.1×SSC、0.1%SDS,更优选68℃,0.1×SSC、0.1%SDS相当的盐浓度和温度下,洗涤一次,更优选两次至三次的条件。
在本发明中“各酶的细胞内活性增强”是指:与野生株(例如大肠杆菌W3110,MG1655株)或亲本株(本发明中指定的酶在所有细胞内活性均未增强的菌株)相比,细胞内的酶活性上升;也包括具有野生株或亲本株所不具有的酶活性。
作为增强细胞内活性的手段,可以列举下述手段及其组合,但不限于此。增强甘油脱氢酶和二羟基丙酮激酶的活性的手段可以采用(1)~(5)中的任何一种,增强这两种酶的活性可以采用相同的手段,也可以采用不同的手段。
(1)通过使用包含编码各蛋白的基因的载体进行转化来增加拷贝数
(2)通过将编码各蛋白的基因整合在染色体上来增加拷贝数
(3)通过对编码各蛋白的基因的表达调节区域进行修饰来增加表达量
(4)通过对影响表达调节的因子进行修饰来增加表达量
(5)通过在编码各蛋白的基因的编码区域内导入突变来增加酶活性
(6)通过提高翻译效率来增大蛋白量
以下,有些情况下会将编码甘油脱氢酶和二羟基丙酮激酶的基因称为目的基因。
(1)通过使用包含编码各蛋白的基因的载体进行转化来增加拷贝数
例如,将包含目的基因的DNA片段与在宿主微生物中发挥功能的载体、优选多拷贝型载体相连接,制作重组DNA,再将其导入微生物进行转化即可。目的基因可以以大肠杆菌、酵母、柠檬酸杆菌或土壤杆菌的染色体DNA为模板,采用PCR法(PCR:聚合酶链式反应;参照White,T.J.et al.,TrendsGenet.5,185(1989))来取得目的基因。其它微生物的目的基因,也可以分别采用以基于该微生物中公知的目的基因或其它种微生物的目的基因或相应的蛋白的序列信息制作的寡核苷酸为引物的PCR方法,或者以基于所述序列信息制作的寡核苷酸为探针的杂交方法,从微生物的染色体DNA或染色体DNA文库中获取。而且,染色体DNA可以采用例如斋藤、三浦的方法(参照H.Saito and K.Miura,Biochem.Biophys.Acta,72,619(1963);生物工学実験書,日本生物工学会编,97-98页,培风馆,1992年)等从作为DNA供体的微生物中制备。
然后,将通过PCR法扩增出的目的基因与能够在宿主微生物的细胞内发挥功能的载体DNA连接来制备重组DNA。作为能够在宿主微生物的细胞内发挥功能的载体,可以列举能够在宿主微生物的细胞内自主复制的载体。
作为能够在属于肠杆菌科的微生物的细胞内自主复制的载体,可以列举pUC19、pUC18、pHSG299、pHSG399、pHSG398、pACYC184、(pHSG、pACYC可以从Takara Bio公司获得),RSF1010(Gene vol.75(2),p271-288,1989),pBR322、pMW219、pMW119(pMW可以从NIIPONGENE公司获得)、pSTV28、pSTV29(Takara Bio公司制造)等。此外,还可以利用噬菌体DNA载体。
为了将这些基因连接到上述载体上来制备重组DNA,需要用与包含目的基因的DNA片段的末端匹配的限制酶来切割载体。连接通常使用T4DNA连接酶等连接酶来进行。DNA的切割、连接以及染色体DNA的制备、PCR、质粒DNA的制备、转化、用作引物的寡核苷酸的设定等的方法,可以采用本领域技术人员熟知的常规方法。这些方法记载于Sambrook,J.,Fritsch,E.F.,and Maniatis,T.,″Molecular Cloning A Laboratory Manual,Second Edition″,Cold Spring Harbor Laboratory Press,(1989)等。
为了将如上述制备的重组DNA导入微生物,可以按照目前为止已经报导的转化方法来进行。例如,可以列举电穿孔法(Canadian Journal ofMicrobiology,43.197(1997))。此外,还可以使用:用氯化钙处理受体菌细胞来增加DNA的透过性的方法(如报道用于大肠杆菌K-12的(Mandel,M.and Higa,A.,J.Mol.Biol.,53,159(1970)),或者由处于增殖阶段的细胞制备感受态细胞并导入DNA的方法(如报道用于枯草芽孢杆菌的(Duncan,C.H.,Wilson,G.A.and Young,F.E.,Gene,1,153(1977))。
(2)通过将编码各蛋白的基因整合在染色体上来增加拷贝数
另一方面,增强各酶的细胞内活性可以通过将目的基因导入到微生物的染色体DNA上从而增加拷贝数来实现。为了将目的基因导入到微生物的染色体DNA上,可以利用染色体DNA上多拷贝存在的序列作为靶标,通过同源重组来进行。作为染色体DNA上多拷贝存在的序列,可以利用重复DNA(repetitive DNA)、存在于转座元件端部的反向重复序列。或者,如特开平2-109985号公报所公开的那样,可以将目的基因加载在转座子上,使其发生转移,从而将其导入染色体DNA上。此外,可以通过采用Red驱动整合法(WO2005/010175)将目的基因导入到染色体上。此外,还可以使用P1噬菌体等噬菌体进行转导,或者利用接合转移载体将目的基因导入到染色体上。这些基因转移到染色体上的事实的确认可以通过以这些基因的一部分为探针、进行Southern杂交来确认。拷贝数得到扩增这一事实的确认可以通过使用与目的基因互补的探针的Southern杂交来进行。拷贝数只要扩增1个拷贝以上可以是任何数目,但编码甘油脱氢酶的基因优选扩增2个拷贝以上、更优选3个拷贝以上、更优选5个拷贝以上;而编码二羟基丙酮激酶的基因优选扩增2个拷贝以上、更优选3个拷贝以上、更优选5个拷贝以上。此外,在导入宿主微生物本来没有的基因时,只要导入1个拷贝以上即可。
(3)通过对编码各蛋白的基因的表达调节区域进行修饰来增加表达量
而且,除了上述的目的基因拷贝数的扩增以外,增强各酶的细胞内活性还可以通过采用国际公布00/18935号小册子中记载的方法、将染色体DNA上或质粒上的这些基因的启动子等表达调节序列更换成强力表达调节序列来实现。例如,lac启动子、trp启动子、trc启动子、λ噬菌体的PR启动子、PL启动子、lpp启动子、T7启动子、tet启动子等是已知的强力启动子。特别是,在本发明中,甘油脱氢酶的扩增适合使用tacM启动子(SEQ IDNO:10)。此外,编码大肠杆菌的二羟基丙酮激酶的dhaK、dhaL、dhaM采取了操纵子结构,可以通过强化dhaK上游的启动子来同时提高3个基因的表达量。
此外,可以在目的基因的启动子区域导入碱基取代等,来将其修饰为更强力的启动子。启动子强度的评价方法和强力启动子的例子记载于Goldstein等的论文(Prokaryotic promoters in biotechnology.Biotechnol.Annu.Rev.,1995,1,105-128)等中。而且,已知核糖体结合位点(RBS)与起始密码子之间的间隔序列,特别是起始密码子紧邻上游的序列中数个核苷酸的取代对mRNA的翻译效率有非常大的影响,可以对其进行修饰。通过这些启动子取代或修饰可以强化目的基因的表达。
将染色体上启动子更换成强启动子,可以通过用通过PCR等扩增出的包含强启动子的DNA转化属于肠杆菌科的微生物,使强启动子与基因组上的野生型启动子之间发生重组,来将基因组上的目的基因上游的启动子更换为强力启动子。利用了这样的同源重组的基因取代,可以利用下述方法:称为“Red驱动整合(Red-driven integration)”的方法(Datsenko,K.A,andWanner,B.L.Proc.Natl.Acad.Sci.U S A.97:6640-6645(2000))、组合使用Red驱动整合法与λ噬菌体来源的切出系统(Cho,E.H.,Gumport,R.I.,Gardner,J.F.J.Bacteriol.184:5200-5203(2002))的方法(参照WO2005/010175号)等使用线性DNA的方法,或者使用含温度敏感型复制起点的质粒的方法等(Datsenko,K.A,and Wanner,B.L.Proc.Natl.Acad.Sci.U S A.97:6640-6645(2000);美国专利第6303383号;特开平05-007491号公报)。
(4)通过对影响表达调节的因子进行修饰来增加表达量
通过对影响表达调节的因子进行修饰来增加表达量,可以通过扩增编码使得编码甘油脱氢酶和编码二羟基丙酮激酶的基因的表达上升的激活物的基因,或者缺失或弱化编码使得这些基因的表达降低的调节物(regulator)的基因来实现。例如,作为编码二羟基丙酮激酶的dhaKLM的激活物,可以列举dhaR(SEQ ID NO:66、GenBank登录号NC_000913的碱基序号1250289..1252208),编码二羟基丙酮激酶的dhaKLM的表达量因dhaR基因的突变而上升(1:EMBO J.2005 Jan 26;24(2):283-93)。此外,ptsI基因(SEQID NO:86GenBank登录号NC_000913的碱基序号2532088..2533815)的破坏也可以导致编码二羟基丙酮激酶的dhaKLM的表达量上升(Microbiology147(2001)247-253)。
(5)通过在编码各蛋白的基因的编码区域内导入突变来增加酶活性
而且,增强甘油脱氢酶和二羟基丙酮激酶的活性还可以通过将能够提高蛋白的比活性的突变、或者提高酶的底物特异性的突变导入目的基因的编码区域内来实现。
编码这样的酶的基因,例如可以通过采用定点突变法对SEQ ID NO:1、3、5或7的碱基序列、表1~4所述的碱基序列中的编码区域进行修饰来获得,所述定点突变使得被编码蛋白中特定位点处的氨基酸残基包含取代、缺失、插入或添加。此外,还可以通过诸如以下的传统公知的突变处理来获得。作为突变处理,用羟胺等对SEQ ID NO:1、3、5或7的碱基序列、表1~4所述的碱基序列、或这些碱基序列中的编码区域的序列进行体外处理的方法,以及对保持该基因的微生物例如属于肠杆菌科的微生物采用紫外线或使用N-甲基-N’-硝基-N-亚硝基胍(NTG)或甲磺酸乙酯(EMS)等在常规突变处理中使用的突变剂来进行处理的方法、易错PCR(Cadwell,R.C.PCR Meth.Appl.2,28(1992))、DNA改组(Stemmer,W.P.Nature 370,389(1994))和StEP-PCR(Zhao,H.Nature Biotechnol.16,258(1998)),利用上述方法通过基因重组人为地在编码甘油脱氢酶和二羟基丙酮激酶的基因中导入突变,来获得编码高活性甘油脱氢酶和二羟基丙酮激酶的基因。这些突变型酶是否编码甘油脱氢酶和二羟基丙酮激酶,例如可以通过下述方法来确定:将这些基因导入具有L-氨基酸生产能力的属于肠杆菌科的微生物,在以甘油为碳源的培养基上进行培养,测定L-氨基酸的生产能力是否提高,或者按上述方法测定酶活性。
(6)通过提高翻译效率来增加蛋白量
为了提高目的蛋白的翻译效率,增加蛋白量,可以通过下述方法来实现:增加与宿主内使用频率低的密码子对应的tRNA、或者对目的基因进行修饰使得其具有与在宿主内使用的宿主的密码子使用频率相应的最适密码子((1989)Gene 85,109-114,(1992)Biochemistry 31,2598-2608.(1993)J.Bacteriol.175,716-722,Protein Expression and Purification 50(2006)49-57)。与非修饰株、野生株相比目的蛋白量的上升,可以通过例如使用抗体通过Western印迹进行检测来确认(Molecular cloning(Cold spring HarborLaboratory Press,Cold spring Harbor(USA),2001))。
本发明的生产方法中使用的微生物,可以是在增强甘油脱氢酶和二羟基丙酮激酶的基础上,还经过修饰而增强了甘油的摄入活性的微生物。甘油的摄入活性是指:将甘油摄取到细胞质(Cytoplasm)的活性,其与一种膜蛋白——甘油易化蛋白(glycerol facilitator)有关。编码甘油易化蛋白的基因可以列举例如大肠杆菌的glpF基因(SEQ ID NO:16:GenBank登录号NC_000913的碱基序号4115268..4116113的互补链)。
编码甘油易化蛋白的基因可以是与SEQ ID NO:16的碱基序列的互补序列或与可从该互补序列制备的探针在严格条件下杂交、且编码具有甘油摄入活性的蛋白的DNA。此外,还可以列举编码SEQ ID NO:17的蛋白的DNA。此外,只要在导入到属于肠杆菌科的微生物中后使甘油摄入能力提高,还可以是相对于SEQ ID NO:17的氨基酸序列全长具有80%以上、优选90%以上、更优选95%、特别优选98%以上的同源性的蛋白。
此外,只要不损害甘油摄入活性,还可以是编码具有在SEQ ID NO:17的氨基酸序列中包含1个或数个氨基酸的取代、缺失、插入或添加等的序列的蛋白的DNA。活性可以采用与上述的增强甘油脱氢酶和二羟基丙酮激酶的方法相似的方法来增强。
甘油摄入的活性可以采用膜蛋白介导的转运测定法(Voegele,R.T.,Sweet,G.D.,and Boos,W.J.Bacteriol.175:1087-1094(1993))来测定。
本发明的生产方法中使用的微生物可以是除了增强甘油脱氢酶和二羟基丙酮激酶、增强甘油的摄入活性之外,还经过修饰而增强了选自丙糖磷酸异构酶、果糖二磷酸醛缩酶、果糖1,6-二磷酸酶、果糖-6-磷酸-醛缩酶中的1种以上的酶活性的微生物。
丙糖磷酸异构酶是指催化将磷酸二羟基丙酮可逆地转化为甘油醛-3-磷酸的反应的酶。(EC:5.3.1.1)
磷酸二羟基丙酮(dihydroxyacetone phosphate)=D-甘油醛-3-磷酸(D-glyceraldehyde 3-phosphate)
“经过修饰而升高了丙糖磷酸异构酶的活性”对应于下述情形:相对于野生株或非修饰株,平均每个细胞的丙糖磷酸异构酶分子数增加、或者丙糖磷酸异构酶平均每个分子的活性提高。优选经过修饰使得丙糖磷酸异构酶的活性与野生株或非修饰株相比较,提高至平均每个菌体150%以上、优选200%以上、更希望为平均每个菌体300%以上。这里,作为对照的属于肠杆菌科的微生物的野生株,包括例如大肠杆菌MG1655株(ATCC No.47076)、和W3110株(ATCC No.27325)、Pantoea ananatis AJ13335株(FERM BP-6615)等。
作为编码丙糖磷酸异构酶的基因,可以列举大肠杆菌来源的tpiA基因(SEQ ID NO:18:GenBank登录号NC_000913的碱基序号4108763..4109530的互补链)。
编码丙糖磷酸异构酶的基因可以是与SEQ ID NO:18的碱基序列的互补序列或与可从该互补序列制备的探针在严格条件下杂交,且编码具有丙糖磷酸异构酶活性的蛋白的DNA。此外,还可以列举编码SEQ ID NO:19的蛋白的DNA。此外,只要在导入属于肠杆菌科的微生物后丙糖磷酸异构酶活性上升,还可以是相对于SEQ ID NO:19的氨基酸序列全长具有80%以上、优选90%以上、更优选95%、特别优选98%以上的同源性的蛋白。
此外,只要不损害丙糖磷酸异构酶活性,还可以是编码具有在SEQ IDNO:19的氨基酸序列中包含1个或数个氨基酸的取代、缺失、插入或添加等的序列的蛋白的DNA。
丙糖磷酸异构酶的活性可以采用Andersen和Cooper的方法(FEBS Lett:4 19-20(1969))来测定。活性可以采用与上述的增强甘油脱氢酶和二羟基丙酮激酶的方法相似的方法来增强。
在本发明中,“果糖二磷酸醛缩酶”是指:可逆地催化从磷酸二羟基丙酮和甘油醛-3-磷酸转化为D-果糖-1,6-二磷酸的以下反应的酶。(EC:4.1.2.13)
磷酸二羟基丙酮(dihydroxyacetone phosphate)(磷酸甘油酮,glyceronephosphate)+D-甘油醛-3-磷酸(D-glyceraldehyde-3-phosphate)=D-果糖-1,6-二磷酸(D-fructose 1,6-bisphosphate)
“经过修饰而升高了果糖二磷酸醛缩酶的活性”对应于下述情形:相对于野生株或非修饰株,平均每个细胞的果糖二磷酸醛缩酶分子数增加,或者果糖二磷酸醛缩酶的平均每个分子的活性提高。优选经过修饰使得果糖二磷酸醛缩酶的活性与野生株或非修饰株相比较,提高至平均每个菌体150%以上、优选200%以上、更希望为平均每个菌体300%以上。这里,作为对照的属于肠杆菌科的微生物的野生株包括例如大肠杆菌MG1655株(ATCCNo.47076)、和W3110株(ATCC No.27325)、Pantoea ananatis AJ13335株(FERM BP-6615)等。
作为编码果糖二磷酸醛缩酶的基因,可以列举大肠杆菌来源的fbaA基因(SEQ ID NO:20:GenBank登录号NC_000913的碱基序号3068187..3069266的互补链)和fbaB基因(SEQ ID NO:72:GenBank登录号NC_000913的碱基序号2175534..2176586的互补链)。
编码果糖二磷酸醛缩酶的基因还可以是与SEQ ID NO:20、72的碱基序列的互补序列或与可从该互补序列制备的探针在严格条件下杂交,且编码具有果糖二磷酸醛缩酶活性的蛋白的DNA。此外,可以列举编码SEQ IDNO:21、73的蛋白的DNA。此外,只要在导入属于肠杆菌科的微生物后果糖二磷酸醛缩酶活性上升,还可以是相对于SEQ ID NO:21的氨基酸序列全长具有80%以上、优选90%以上、更优选95%、特别优选98%以上的同源性的蛋白。
此外,只要不损害果糖二磷酸醛缩酶活性,还可以是编码具有在SEQ IDNO:21、73的氨基酸序列中包含1个或数个氨基酸的取代、缺失、插入或添加等的序列的蛋白的DNA。
果糖二磷酸醛缩酶的活性可以采用Richard和Rutter的方法(J.Biol.Chem.236,3177-3184)来测定。活性可以采用与上述增强甘油脱氢酶和二羟基丙酮激酶的方法相似的方法来增强。
果糖1,6-二磷酸酶是指:可逆地催化将D-果糖-1,6-二磷酸转化为D-果糖6-磷酸的以下反应的酶。(EC:3.1.3.11)
D-果糖-1,6-二磷酸(D-fructose 1,6-bisphosphate)+H2O=D-果糖6-磷酸(D-fructose 6-phosphate)+磷酸(phosphate)
“经过修饰而升高了果糖1,6-二磷酸酶的活性”对应于下述情形:相对于野生株或非修饰株,平均每个细胞的果糖1,6-二磷酸酶分子数增加,或者果糖1,6-二磷酸酶的平均每个分子的活性提高。优选经过修饰而使得果糖1,6-二磷酸酶的活性与野生株或非修饰株相比较,平均每个菌体提高至150%以上、优选200%以上、更希望为平均每个菌体300%以上。这里,作为对照的属于肠杆菌科的微生物的野生株包括例如大肠杆菌MG1655株(ATCCNo.47076)、和W3110株(ATCC No.27325)、Pantoea ananatis AJ13335株(FERM BP-6615)等。
作为编码果糖1,6-二磷酸酶的基因,可以列举大肠杆菌来源的glpX基因(SEQ ID NO:22:GenBank登录号NC_000913的碱基序号4112592..4113602的互补链)、fbp基因(SEQ ID NO:82:GenBank登录号NC_000913的碱基序号4452634..4453632)、ybhA基因(SEQ ID NO:84:GenBank登录号NC_000913的碱基序号796836..7976554)。编码果糖1,6-二磷酸酶的基因还可以是与SEQ ID NO:22、82、84的碱基序列的互补序列或与可从该互补序列制备的探针在严格条件下杂交,且编码具有果糖1,6-二磷酸酶活性的蛋白的DNA。此外,可以列举编码SEQ ID NO:23、83、85的蛋白的DNA。此外,只要在导入属于肠杆菌科的微生物后果糖二磷酸醛缩酶活性上升,还可以是与SEQ ID NO:23、83、85的氨基酸序列全体具有80%以上、优选90%以上、更优选95%、特别优选98%以上的同源性的蛋白。
此外,只要不损害果糖1,6-二磷酸酶活性,还可以是编码具有在SEQ IDNO:23、83、85的氨基酸序列中包含1个或数个氨基酸的取代、缺失、插入或添加等的序列的蛋白的DNA。
果糖1,6-二磷酸酶的活性可以采用中岛等的方法(蛋白質核酸酵素22,1585-1589)来测定。活性可以采用与上述的增强甘油脱氢酶和二羟基丙酮激酶的相似的方法来增强。
在本发明中,“果糖-6-磷酸醛缩酶”是指:可逆地催化将二羟基丙酮转化为果糖-6-磷酸的以下反应的酶。
D-果糖-6-磷酸(D-fructose-6-phosphate)=二羟基丙酮(dihydroxyacetone)+D-甘油醛-3-磷酸(D-glyceraldehyde-3-phosphate)“经过修饰而升高了果糖-6-磷酸醛缩酶的活性”对应于下列情形:相对于野生株或非修饰株,平均每个细胞的果糖-6-磷酸醛缩酶分子数增加、或者果糖-6-磷酸醛缩酶平均每个分子的活性提高。优选经过修饰使得果糖-6-磷酸醛缩酶的活性与野生株或非修饰株相比较,平均每个菌体提高至150%以上、优选200%以上、更希望为平均每个菌体300%以上。这里,作为对照的属于肠杆菌科的微生物的野生株包括例如大肠杆菌MG1655株(ATCCNo.47076)、和W3110株(ATCC No.27325)、Pantoea ananatis AJ13335株(FERM BP-6615)等。
作为编码果糖-6-磷酸醛缩酶的基因,可以列举:编码I型醛缩酶的大肠杆菌来源的fsaA基因(SEQ ID NO:68:GenBank登录号NC_000913的碱基序号862865..863527)、编码II型醛缩酶的fsaB基因(talC基因)(SEQ IDNO:70:GenBank登录号NC_000913的碱基序号4137069..4137731的互补链)。
编码果糖-6-磷酸醛缩酶的基因还可以是与SEQ ID NO:68、70的碱基序列的互补序列或与可由该互补序列制备的探针在严格条件下杂交、且编码具有果糖-6-磷酸醛缩酶活性的蛋白的DNA。此外,还可以列举编码SEQID NO:69、71的蛋白的DNA。此外,只要在导入属于肠杆菌科的微生物后果糖-6-磷酸醛缩酶活性上升,还可以是与SEQ ID NO:69、71的氨基酸序列全长具有80%以上、优选90%以上、更优选95%、特别优选98%以上的同源性的蛋白。
此外,只要不损害果糖-6-磷酸醛缩酶活性,还可以是编码具有在SEQ IDNO:69、71的氨基酸序列中包含1个或数个氨基酸的取代、缺失、插入或添加等的序列的蛋白的DNA。
果糖-6-磷酸醛缩酶的活性可以采用Schurmann M,Sprenger GA等的方法(J Biol Chem.2001 Apr 6;276(14):11055-61)来测定。活性可以采用与上述的增强甘油脱氢酶和二羟基丙酮激酶的方法相似的方法来增强。
本发明的生产方法中使用的微生物,除了甘油脱氢酶和二羟基丙酮激酶增强、甘油摄入活性增强、选自丙糖磷酸异构酶、果糖二磷酸醛缩酶、果糖1,6-二磷酸酶、果糖-6-磷酸醛缩酶中的1种以上的酶活性增强之外,优选还经过修饰而降低了甘油激酶和/或膜结合型甘油-3-磷酸脱氢酶的活性。
在本发明中,“甘油激酶”是指:可逆地催化从甘油和ATP生成甘油3-磷酸和ADP的以下反应的酶。(EC 2.7.1.30)
ATP+甘油(glycerol)=ADP+sn-甘油3-磷酸(sn-glycerol 3-phosphate)“经过修饰而降低了甘油激酶的活性”对应于下述情况:相对于野生株或非修饰株,平均每个细胞的甘油激酶分子数减少,或者甘油激酶的平均每个分子的活性降低。优选经过修饰使得与野生株或非修饰株相比甘油激酶的活性平均每个菌体降低至70%以下、优选50%以下、更希望30%以下、最优选平均每个菌体20%以下,也可以使酶活性丧失。酶活性可以通过降低编码该酶的基因的表达量来降低。基因表达量的降低包括由该基因转录的mRNA的转录量降低、和该mRNA的翻译量降低。
此外,使得酶蛋白分子完全不生成、或者使酶蛋白平均每个分子的活性降低或丧失,可以通过破坏编码该酶的基因来实现。这里,作为对照的属于肠杆菌科的微生物的野生株包括例如大肠杆菌MG1655株(ATCCNo.47076)、和W3110株(ATCC No.27325)、Pantoea ananatis AJ13335株(FERM BP-6615)等。
作为编码甘油激酶的基因,可以列举大肠杆菌来源的glpK基因(SEQ IDNO:24:GenBank登录号NC_000913的碱基序号4113737..4115245的互补链)。甘油激酶的酶活性可以采用Thorner&Paulus的方法来测定(TheEnzymes(3rd ed.)8,487-508)
在本发明中,“膜结合型甘油-3-磷酸脱氢酶”是指:可逆地催化从甘油-3-磷酸到磷酸二羟基丙酮的氧化反应的酶,即催化以下反应的酶。
sn-甘油-3-磷酸(sn-glycerol-3P)+泛醌(ubiquinone)=磷酸二羟基丙酮(dihydroxyaceton-P)+泛醇(ubiquinol)(EC:1.1.99.5)
“经过修饰而降低了膜结合型甘油-3-磷酸脱氢酶的活性”对应于下列情形:相对于野生株或非修饰株,平均每个细胞的膜结合型甘油-3-磷酸脱氢酶分子数减少,或者膜结合型甘油-3-磷酸脱氢酶的平均每个分子的活性降低。优选经过修饰使得与野生株或非修饰株相比,膜结合型甘油-3-磷酸脱氢酶的活性平均每个菌体降低至70%以下、优选50%以下、更希望30%以下,也可以使酶活性丧失。酶活性可以通过降低编码该酶的基因的表达量来降低。这里,作为对照的属于肠杆菌科的微生物的野生株包括例如大肠杆菌MG1655株(ATCC No.47076)、和W3110株(ATCC No.27325)、Pantoeaananatis AJ13335株(FERM BP-6615)等。
膜结合型的甘油-3-磷酸脱氢酶由glpABC操纵子、glpD基因编码。作为大肠杆菌的glpA基因,可以列举SEQ ID NO:26(GenBank登录号NC_000913的碱基序号2350669..2352297);作为glpB基因,可以列举SEQID NO:28(GenBank登录号NC_000913的碱基序号2352287..2353546);作为glpC基因,可以列举SEQ ID NO:30(GenBank登录号NC_000913的碱基序号2353543..2354733);作为glpD基因,可以列举SEQ ID NO:32(GenBank登录号NC_000913的碱基序号3560036..3561541)。
上述甘油激酶、甘油-3-磷酸脱氢酶等目的酶的活性降低,可以通过
(1)通过在编码目的酶的基因的编码区域内导入突变来导致酶活性降低或丧失、
(2)通过对编码目的酶的基因的表达调节序列进行修饰来导致酶活性降低或丧失,
来实现。
(1)通过在编码目的酶的基因的编码区域内导入突变来导致酶活性降低或丧失
编码目的酶的基因的编码区域内的突变导入可以通过下述方法实现:通过基因重组,在染色体上的编码酶的区域中导入氨基酸取代(错义突变),或导入终止密码子(无义突变),或者导入一~二个碱基添加或缺失引起的移框突变(Journal of Biological Chemistry 272:8611-8617(1997)Proceedings ofthe National Academy of Sciences,USA 95 5511-5515(1998),Journal ofBiological Chemistry 266,20833-20839(1991))。此外,也可以通过缺损编码区域内的基因的一部分或全部来实现。具体地,可以通过在SEQ ID NO:24、26、28、30、32的DNA的一部分中导入突变、或者缺失DNA的一部分或者全部来实现。
这些突变导入也可以通过下述方式来降低或缺损酶活性:构建编码区域中有缺失或导入了突变的突变酶的基因,通过同源重组等用该基因将染色体上的正常基因取代为突变型基因;将转座子、IS因子等导入该基因。
例如,为了通过基因重组导入上述能够降低或缺损的酶活性的突变,可以采用以下的方法。可以对目的基因的部分序列进行修饰,制作不产生正常发挥功能的酶的突变型基因,用含该基因的DNA转化属于肠杆菌科的微生物,使突变型基因与基因组上的基因间发生重组,从而将基因组上的目的基因取代为突变型。利用了这样的同源重组的基因取代,可以利用下述方法:称为“Red驱动整合(Red-driven integration)”的方法(Datsenko,K.A,and Wanner,B.L.Proc.Natl.Acad.Sci.U S A.97:6640-6645(2000))、组合使用Red驱动整合法与λ噬菌体来源的切出系统(Cho,E.H.,Gumport,R.I.,Gardner,J.F.J.Bacteriol.184:5200-5203(2002))的方法(参照WO2005/010175号)等使用线性DNA的方法,或者使用含温度敏感型复制起点的质粒的方法等(Datsenko,K.A,and Wanner,B.L.Proc.Natl.Acad.Sci.U S A.97:6640-6645(2000);美国专利第6303383号;特开平05-007491号公报)。此外,诸如上述的通过利用了同源重组的基因取代进行的定点突变导入,还可以使用在宿主上不具有复制能力的质粒来进行。此外,酶活性的降低或丧失还可以是通过X射线或紫外线照射、或者利用N-甲基-N’-硝基-N-亚硝基胍等突变剂进行的常规突变处理,在目的基因的编码区域内导入突变的修饰。
(2)通过对编码目的酶的基因的表达调节序列进行修饰来导致酶活性降低或丧失
通过对编码目的酶的基因的表达调节序列进行修饰来导致酶活性降低或丧失,可以这样实现:在染色体DNA上的启动子、SD序列等表达调节序列中导入突变来降低表达量;扩增编码能够降低这些基因的表达的调节物的基因;缺失或弱化编码能够提高这些基因的表达的激活物的基因。启动子强度的评价方法和强力启动子的例子记载于Goldstein等的论文(Prokaryotic promoters in biotechnology.Biotechnol.Annu.Rev.,1995,1,105-128)等中。而且,已知核糖体结合位点(RBS)与起始密码子之间的间隔序列、特别是起始密码子紧邻上游的序列中数个核苷酸的取代对mRNA的翻译效率有非常大的影响,可以对它们进行修饰。特别是,glpA、B、C基因采取操纵子结构,因此,可以通过在glpA上游的启动子区域等表达调节区域中导入突变来降低表达量。
<2>本发明的生产方法
本发明的方法是以在培养基中包含甘油的培养基中培养上述的属于肠杆菌科、且具有L-氨基酸生产能力的细菌,并在培养基中或菌体内生成并蓄积L-氨基酸,从该培养基或菌体收集L-氨基酸为特征的L-氨基酸的生产方法。这里,本发明的方法可以采用分批培养(batch culture)、流加培养(Fed-batch culture)、连续培养法(continuous culture)中的任何方法,培养基中的甘油可以包含在初始培养基中,也可以包含在流加培养基中,或者包含于这两者中。
这里,在本发明中,上述流加培养是指下述培养方法:将培养基连续地或间歇地添加至培养容器中,并且在培养完成之前不从容器中取出培养基。此外,连续培养指下述培养方法:连续地或间歇地将培养基流加至培养容器中,同时从容器中取出培养基(通常而言,等于所流加的培养基的量)。“初始培养基”的意思是在流加培养或连续培养中,流加流加培养基之前的分批培养中所用的培养基(培养开始时的培养基);“流加培养基”的意思是在进行流加培养或连续培养时,提供至发酵罐的培养基。此外,分批培养(bachculture)是指:每批一次准备新的培养基,将菌株接种到其中,并且直至收获都不加入培养基。
本发明的培养基中包含的甘油可以是单独的碳源,还可以使用除甘油以外还添加了其它碳源的混合培养基。优选的是葡萄糖、果糖、蔗糖、乳糖、半乳糖、废糖蜜、淀粉水解物或由生物质的水解得到的糖液等糖类,乙醇等醇类,富马酸、柠檬酸、琥珀酸等有机酸类。在使用混合培养基时,培养基中的甘油的比例希望包含50%以上、60%以上、优选70%以上、更优选80%以上、特别优选90%以上。特别优选的是,所利用的甘油是作为生物柴油的副产品得到的。(Mu Y,et al,Biotechnol Lett.,28,1755-91759(2006),Haas MJ,et al;Bioresour Technol.97,4,671-8678(2006))
可以使用常规的培养基,其中除碳源外,还含有氮源、无机离子和视需要而定的其它有机成分作为培养基中添加的其它成分。
作为本发明的培养基中包含的氮源,可以使用氨气、硫酸铵、碳酸铵、氯化铵、磷酸铵、醋酸铵、尿素等铵盐或硝酸盐等,pH调整中使用的氨气、氨水也可作为氮源利用。此外,还可以利用蛋白胨、酵母提取物、肉提取物、麦芽提取物、玉米浆、大豆水解物等。培养基中也以只含这些氮源中的1种,也可以含有2种以上。这些氮源可以用于初始培养基,也可以用于流加培养基。此外,初始培养基、流加培养基中可以使用相同的氮源,也可以将流加培养基的氮源改变为与初始培养基不同。
本发明的培养基中,除了碳源、氮源、硫之外,优选还包含磷酸源、硫源。作为磷酸源,可以利用磷酸二氢钾、磷酸氢二钾、焦磷酸等磷酸多聚物等。此外,作为硫源,只要包含硫原子即可,优选硫酸盐、硫代硫酸盐、亚硫酸盐等硫酸盐,半胱氨酸、胱氨酸、谷胱甘肽等含硫氨基酸,这其中优选硫酸铵。
此外,培养基除碳源、氮源、硫之外,还可以包含生长促进因子(具有生长促进效果的营养素)。能够使用的生长促进因子包括痕量金属、氨基酸、维生素、核酸以及含有前述物质的蛋白胨、酪蛋白氨基酸、酵母提取物、大豆蛋白降解产物等。作为痕量金属,可以列举铁、锰、镁、钙等;作为维生素,可以列举维生素B1、维生素B2、维生素B6、烟酸、烟酰胺、维生素B12等。这些生长促进因子可以包含在初始培养基中,也可以包含在流加培养基中。
此外,在使用其生长需求氨基酸等的营养缺陷型突变株时,优选在培养基中添补所需求的营养素。特别是,可在本发明中使用的L-赖氨酸生产菌中大多如后述地强化了L-赖氨酸生物合成途径,而弱化了L-赖氨酸分解能力,因此,优选添加选自L-苏氨酸、L-高丝氨酸、L-异亮氨酸、L-甲硫氨酸中的1种或2种以上。
初始培养基和流加培养基的培养基组成可以相同,也可以不同。此外,初始培养基与流加培养基的硫浓度可以相同,也可以不同。而且,当流加培养基的流加分多个阶段进行时,各流加培养基的组成可以相同,也可以不同。
培养优选在发酵温度20至45℃,特别优选33至42℃的通气条件下进行。这里,将氧浓度调节到5至50%,优选至大约10%。此外,优选将pH控制在5至9进行通气培养。如果在培养期间pH降低,例如,可添加碳酸钙或碱例如氨气和氨水以中和培养物。在这些条件下,进行优选大约10至120小时的培养,从而在培养液中积累显著量的L-氨基酸。蓄积的L-氨基酸的浓度只要是高于野生型菌株中的浓度并且在该浓度下能够将L-氨基酸从培养基收集和回收即可,可以是50g/L或更高,理想的是75g/L或更高,更理想的是100g/L或更高。
就培养结束后从培养液中收集L-氨基酸的方法而言,可以按照公知的回收方法来进行。例如,通过采用离心分离等从培养液中除去菌体、然后进行浓缩晶析来进行收集。
在本发明中,为了确保高于一定水平的L-氨基酸蓄积,微生物的培养可以分为种子培养(seed culture)和主培养(main culture)来进行,种子培养可通过使用培养瓶等的摇动培养或通过分批培养来进行,而主培养可以以流加培养或连续培养的形式进行。或者,种子培养和主培养均可以以分批培养的方式进行。
当根据本发明进行流加培养或连续培养时,可间歇地流加流加培养基,使甘油或其它碳源的流加暂时停止。优选的是,停止流加培养基供给的时间为流加进行时间的至多30%,理想的是至多20%,特别理想的是至多10%。当间歇流加流加培养液时,可添加一定时间的流加培养基,并如下控制第二次及以后的添加:在某个添加期前面的添加停止期中发酵培养基内碳源耗尽时pH的上升和溶氧浓度的上升通过计算机被检测到时,即开始添加;并且因此培养罐(culture tank)中的基质浓度可自动地保持在低水平(美国专利5,912,113号说明书)。
流加培养中使用的流加培养基,优选含甘油或其它碳源、以及具有生长促进效果的营养素(生长促进因子)的培养基,可以进行控制,使得发酵培养基中的甘油浓度、其它碳源的浓度在一定水平以下。作为其它碳源,优选葡萄糖、蔗糖、果糖;作为生长促进因子,优选氮源、磷酸、氨基酸等。作为氮源,可以使用氨气、硫酸铵、碳酸铵、氯化铵、磷酸铵、醋酸铵、尿素等铵盐或硝酸盐等。此外,作为磷酸源,可以使用磷酸二氢钾、磷酸氢二钾,作为氨基酸,优选在使用营养缺陷型突变株时添补要求的营养素。此外,流加培养基可以是1种,也可以混合2种以上的培养基。在使用2种以上流加培养基时,这些培养基可以混合起来通过1个发酵罐流加,也可以通过多个发酵罐流加。
在将连续培养方法用于本发明时,可以同时取出和添加;也可以取出部分后再添加。此外,所述方法也可以是对菌体进行再利用的连续培养方法,其中将包含L-氨基酸和细胞的培养液取出,并且仅将细胞再循环回发酵罐(参照法国专利号2669935说明书)。作为连续或间歇流加营养源的方法,使用与流加培养中所用的相同方法。
再利用菌体的连续培养方法是如下所述的方法:在氨基酸浓度达到预定水平时,将发酵培养基间歇或连续地取出,仅取出L-氨基酸,并且将包含菌体的过滤残余物再循环到发酵罐中,这种方法可通过参考例如法国专利号2669935说明书来实施。
其中,将培养液间歇取出时,可以在L-氨基酸浓度达到预定浓度时取出部分L-氨基酸,并且新添加培养基以继续培养。此外,至于添加的培养基的量,优选进行设定使其与最终取出之前培养液的量为等量。这里,“等量”的意思是取出之前培养液量的大约93~107%的量。
在将培养液连续取出时,优选在流加营养培养基的同时或之后开始取出。例如,取出开始于添加开始之后的5小时之内,优选3小时之内,更优选1小时之内。此外,至于培养液的取出量,优选取出的量和流加的量为等量。
<3>作为本发明的微生物的亲本株使用的微生物
在本发明中,可以使用下述细菌作为亲本株,并按上述方法来赋予本发明的性质,所述细菌属于肠杆菌科,且具有能够以甘油为碳源进行代谢的L-氨基酸生产能力。
这里,肠杆菌科细菌中包含属于埃希氏菌、肠杆菌、欧文氏菌、克雷伯氏菌、泛菌、光状杆菌(Photorhabdus)、普罗威登斯菌(Providencia)、沙门氏菌、沙雷氏菌、志贺氏菌、摩根氏菌、耶尔森氏菌等属的细菌。特别是优选通过NCBI(National Center for Biotechnology Information)的数据库(http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=91347)中采用的分类法分类在肠杆菌科的细菌。
关于“属于埃希氏菌属的细菌”,不意在限制,是指该细菌按照微生物学专家知晓的分类方法分类在埃希氏菌属。在本发明中使用的属于埃希氏菌属的细菌的例子包含大肠杆菌(E.coli),但不限于此。
对于可在本发明中使用的属于埃希氏菌属的细菌而言,不意在限制,包含例如Bachmann等的著作的表1(Bachmann,B.J.1996.p.2460-2488.InF.D.Neidhardt(编),Escherichia coli and Salmonella Cellular and MolecularBiology/Second Edition,American Society for Microbiology Press,Washington,D.C)中描述的系统。具体地,可以列举原型野生株K12株来源的大肠杆菌W3110(ATCC 27325)、大肠杆菌MG1655(ATCC 47076)等。
这些菌株可以从例如美国典型培养物保藏中心(地址12301 ParklawnDrive,Rockville,Maryland 20852,United States of America)通过分售获得。即,对各菌株赋予了对应的注册号,可以利用该注册号通过分售获得。对应于各菌株的注册号见美国典型培养物保藏中心的目录。
属于泛菌属的细菌是指:该细菌通过微生物学专家知晓的分类被分类在泛菌属。成团肠杆菌的某些种,最近基于16S rRNA碱基序列分析等被重新分类为成团泛菌、Pantoea ananatis、斯氏泛菌((Pantoea stewartii))等(Int.J.Syst.Bacteriol.1993.43:162-173)。在本发明中,属泛菌属的细菌也包含这样重新分类在泛菌属的细菌。
在本发明中,具有L-氨基酸生产能力的细菌是指:具有在培养基中培养后,生产L-氨基酸,并将L-氨基酸分泌到培养基中的能力的细菌。此外,优选是指:能够在培养基中蓄积优选0.5g/L以上、更优选1.0g/L以上的量的目的L-氨基酸的细菌。L-氨基酸包括L-丙氨酸、L-精氨酸、L-天冬酰胺、L-天冬氨酸、L-半胱氨酸、L-谷氨酸、L-谷氨酰胺、甘氨酸、L-组氨酸、L-异亮氨酸、L-亮氨酸、L-赖氨酸、L-甲硫氨酸、L-苯丙氨酸、L-脯氨酸、L-丝氨酸、L-苏氨酸、L-色氨酸、L-酪氨酸和L-缬氨酸。特别优选L-苏氨酸和L-赖氨酸。
在本发明的亲本株中,可以使用目前为止报导的L-氨基酸生产菌,只要能够同化甘油即可。以下,对可在本发明的方法中使用的各L-氨基酸生产菌进行说明。
L-苏氨酸生产菌
作为L-苏氨酸生产菌或用于衍生L-苏氨酸生产菌的亲本株的例子,可以列举:大肠杆菌TDH-6/pVIC40(VKPM B-3996)(美国专利第5,175,107号、美国专利第5,705,371号)、大肠杆菌472T23/pYN7(ATCC 98081)(美国专利第5,631,157号)、大肠杆菌NRRL-21593(美国专利第5,939,307号)、大肠杆菌FERM BP-3756(美国专利第5,474,918号)、大肠杆菌FERM BP-3519和FERM BP-3520(美国专利第5,376,538号)、大肠杆菌MG442(Gusyatiner etal.,Genetika(俄语),14,947-956(1978))、大肠杆菌VL643和VL2055(EP1149911 A)等属于埃希氏菌属的菌株,但不限于此。
TDH-6菌株在thrC基因有缺陷,是蔗糖同化型,并且ilvA基因有泄漏突变(leaky mutation)。该菌株的rhtA基因也有突变,该突变赋予对高浓度苏氨酸或高丝氨酸的抗性。B-3996菌株包含pVIC40,pVIC40是通过将含有突变的thrA基因的thrA*BC操纵子插入到由RSF1010获得的载体中而获得的。这种突变的thrA基因编码的天冬氨酸激酶高丝氨酸脱氢酶I对苏氨酸的反馈抑制基本上已解除。B-3996菌株在1987年11月19日被保藏于All-Union Scientific Center of Antibiotics(全联盟抗生素科学中心)(Nagatinskaya Street 3-A,117105 Moscow,Russia),保藏号为RIA 1867。该菌株也在1987年4月7日保藏于俄罗斯国立工业微生物保藏中心(RussianNational Collection of Industrial Microorganisms)(VKPM),保藏号为B-3996。
还可以使用大肠杆菌VKPM B-5318(EP 0593792B)作为L-苏氨酸生产菌或用于衍生L-苏氨酸生产菌的亲本株。B-5318菌株是异亮氨酸原养型的,并且pVIC40质粒中的苏氨酸操纵子的调控区域被置换为温度敏感的λ噬菌体C1阻遏物和PR启动子。VKPM B-5318菌株在1990年5月3日保藏于俄罗斯国立工业微生物保藏中心(VKPM,1 Dorozhny proezd.,1Moscow117545,Russia),保藏号为VKPM B-5318。
优选地,本发明的细菌进一步经过修饰而增强了一种或多种下列基因的表达:
-突变thrA基因,其编码对苏氨酸的反馈抑制具有抗性的天冬氨酸激酶高丝氨酸脱氢酶I;
-thrB基因,其编码高丝氨酸激酶;
-thrC基因,其编码苏氨酸合酶(threonine synthase);
-rhtA基因,其编码推定的跨膜蛋白;
-asd基因,其编码天冬氨酸-β-半醛脱氢酶;
-aspC基因,其编码天冬氨酸氨基转移酶(天冬氨酸转氨酶)。
编码天冬氨酸激酶高丝氨酸脱氢酶I的大肠杆菌thrA基因已经阐明(核苷酸编号337至2799,GenBank登录号NC_000913.2,gi:49175990)。thrA基因位于大肠杆菌K-12染色体上thrL基因和thrB基因之间。大肠杆菌的编码高丝氨酸激酶的thrB基因已经阐明(核苷酸编号2801至3733,GenBank登录号NC_000913.2,gi:49175990)。thrB基因位于大肠杆菌K-12染色体上的thrA基因和thrC基因之间。大肠杆菌的编码苏氨酸合酶的thrC基因已经阐明(核苷酸编号3734至5020,GenBank登录号NC_000913.2,gi:49175990)。thrC基因位于大肠杆菌K-12染色体上thrB基因和yaaX开放阅读框之间。所有这3个基因作为一个单独的苏氨酸操纵子发挥功能。为增加苏氨酸操纵子的表达,优选从操纵子中将影响转录的弱化子区域去除(WO2005/049808,WO2003/097839)。
编码对苏氨酸的反馈抑制有抗性的天冬氨酸激酶高丝氨酸脱氢酶I的突变thrA基因,以及thrB和thrC基因可以作为一个操纵子从熟知的pVIC40质粒中获得,该质粒存在于苏氨酸生产株大肠杆菌VKPM B-3996菌株中。pVIC40质粒在美国专利5,705,371中有详述。
rhtA基因存在于大肠杆菌染色体上的18min处,靠近编码谷氨酰胺转运系统的成分的glnHPQ操纵子。rhtA基因与ORF1(ybiF基因,核苷酸编号764至1651,GenBank登录号AAA218541,gi:440181)相同,位于pexB与ompX基因之间。已将表达由ORF1编码的蛋白的序列命名为rhtA基因(rht:对高丝氨酸和苏氨酸抗性)。此外,已经证明rhtA23突变是在相对于ATG起始密码子的-1位置用G→A取代(ABSTRACTS of the 17thInternational Congress of Biochemistry and Molecular Biology in conjugationwith Annual Meeting of the American Society for Biochemistry and MolecularBiology(第17界国际生物化学与分子生物学大会暨美国生物化学与分子生物学协会年会摘要),San Francisco,California,1997年8月24-29日,摘要号457,EP 1013765A)。
大肠杆菌的asd基因已经阐明(核苷酸编号3572511至3571408,GenBank登录号NC_000913.1,gi:16131307),并且可以根据该基因的碱基序列利用引物通过PCR(参见White,T.J.等,Trends Genet.,5,185(1989))来获得。其它微生物的asd基因可以通过类似方式获得。
此外,大肠杆菌的aspC基因已经阐明(核苷酸编号983742至984932,GenBank登录号NC_000913.1,gi:16128895),并且可以通过PCR获得。其它微生物的aspC基因可以通过类似方式获得。
L-赖氨酸生产菌
属于埃希氏菌属的L-赖氨酸生产菌的实例包括对L-赖氨酸类似物具有抗性的突变株。L-赖氨酸类似物抑制属于埃希氏菌属细菌的生长,但是当L-赖氨酸共存于培养基中时这种抑制会被全部或部分解除。L-赖氨酸类似物的实例包括、但不限于,氧化赖氨酸,赖氨酸氧肟酸(lysine hydroxamate),S-(2-氨乙基)-L-半胱氨酸(AEC),γ-甲基赖氨酸,α-氯代己内酰胺等。对属于埃希氏菌属的细菌进行常规的人工诱变处理可以得到对这些赖氨酸类似物具有抗性的突变株。可用于L-赖氨酸生产的菌株的具体实例包括大肠杆菌AJ11442(FERM BP-1543,NRRL B-12185;参见美国专利第4,346,170号)和大肠杆菌VL611。在这些微生物中,L-赖氨酸对天冬氨酸激酶的反馈抑制已被解除。
WC196菌株可用作L-赖氨酸生产菌。这种菌株是通过赋予源自大肠杆菌K-12的W3110菌株以AEC抗性选育而成的。将该菌株命名为大肠杆菌AJ13069,并于1994年12月6日保藏在日本工业技术院生命工学工业技术研究所(现为独立行政法人产业技术综合研究所特许微生物保藏中心,邮箱305-8566,日本国茨城县筑波市东1丁目1番地中央第6),取得保藏号FERMP-14690。其后根据布达佩斯条约的规定于1995年9月29日转为国际保藏,并取得保藏号FERM BP-5252(美国专利第5,827,698号)。
L-赖氨酸生产菌或用于衍生L-赖氨酸生产菌的亲本株的实例还包括编码L-赖氨酸生物合成酶的1种以上的基因的表达增强的菌株。相关基因的实例包括但不限于,二氢吡啶二羧酸合酶基因(dapA)、天冬氨酸激酶基因(lysC)、二氢吡啶二羧酸还原酶基因(dapB)、二氨基庚二酸脱羧酶基因(lysA)、二氨基庚二酸脱氢酶基因(ddh)(美国专利6,040,160)、磷酸烯醇丙酮酸羧化酶基因(ppc)、天冬氨酸半醛脱氢酶基因(asd)和天冬氨酸酶基因(aspA)(EP1253195 A)。此外,亲本菌株中涉及能量效率(energy efficiency)的基因(cyo)(EP 1170376 A)、编码烟酰胺核苷酸转氢酶的基因(pntAB)(美国专利5,830,716)、ybjE基因(WO2005/073390)、编码谷氨酸脱氢酶的基因(gdhA)(Gene23:199-209(1983))或它们的组合的表达水平增加。而且,括号内为该基因的简记符号。
已知大肠杆菌来源的野生型二氢吡啶二羧酸合成酶受L-赖氨酸的反馈抑制,大肠杆菌来源的野生型天冬氨酸激酶受L-赖氨酸的抑制和反馈抑制。因此,在使用dapA基因和lysC基因时,优选这些基因是不受L-赖氨酸的反馈抑制的突变型基因。
作为编码不受L-赖氨酸的反馈抑制的突变型二氢吡啶二羧酸合成酶的DNA,可以列举编码具有第118位的组氨酸残基被取代成酪氨酸残基的序列的蛋白的DNA。此外,作为编码不受L-赖氨酸的反馈抑制的突变型天冬氨酸激酶的DNA,可以列举编码具有第352位的苏氨酸残基被取代为异亮氨酸残基、第323位的甘氨酸残基被取代为天冬酰胺残基、第318位的甲硫氨酸被取代为异亮氨酸的序列的AKIII的DNA(关于这些突变体,可参见美国专利第5661012号和第6040160号说明书)。突变型DNA可以通过利用PCR等的定点突变方法获得。
而且,作为包含编码突变型突变型二氢吡啶二羧酸合成酶的突变型dapA和编码突变型天冬氨酸激酶的突变型lysC的质粒,已知有广宿主域质粒RSFD80、pCAB1、pCABD2(美国专利第6040160号说明书)。用RSFD80转化的大肠杆菌JM109株(美国专利第6040160号说明书)命名为AJ12396,该株于1993年10月28日保藏在日本通产省工业技术院生命工学工业技术研究所(现为独立行政法人产业技术综合研究所专利生物保藏中心),保藏号FERM P-13936;并于1994年11月1日转为基于布达佩斯条约的国际保藏,保藏号FERM BP-4859。RSFD80可采用公知方法从AJ12396株获取。
作为L-赖氨酸生产菌或用于衍生L-赖氨酸生产菌的亲本株的例子,可列举催化下述反应的酶的活性减少或缺损的菌株,所述反应通过从L-赖氨酸生物合成途径分支,而生成除L-赖氨酸外的化合物。作为催化通过从L-赖氨酸生物合成途径分支而生成除L-赖氨酸外的化合物的反应的酶的例子,可以列举高丝氨酸脱氢酶、赖氨酸脱羧酶(美国专利第5,827,698号)、和苹果酸酶(WO2005/010175)。这里,为了降低或缺损赖氨酸脱羧酶活性,优选降低编码赖氨酸脱羧酶的cadA基因和ldcC基因这两者的表达(国际公布第WO2006/038695号小册子)。
L-半胱氨酸生产菌
L-半胱氨酸生产菌或用于衍生L-半胱氨酸生产菌的亲本株的实例包括、但不限于下述属于埃希氏菌属的菌株,如用编码反馈抑制抗性丝氨酸乙酰转移酶的不同cysE等位基因转化的大肠杆菌JM15(美国专利6,218,168,俄罗斯专利申请2003121601),具有过表达编码适于向细胞分泌有毒物质的蛋白的基因的大肠杆菌W3110(美国专利5,972,663),半胱氨酸脱巯基酶(cysteine desulfohydrase)活性减少的大肠杆菌菌株(JP11155571A2);由cysB基因编码的半胱氨酸调节子的正向转录调控物活性增强的大肠杆菌W3110(WO0127307A1),等等。
L-亮氨酸生产菌
L-亮氨酸生产菌或用于衍生L-亮氨酸生产菌的亲本株的实例包括、但不限于下述属于埃希氏菌属的菌株,如对亮氨酸有抗性的大肠杆菌菌株(例如,菌株57(VKPM B-7386,美国专利6,124,121))或对亮氨酸类似物(包括β-2-噻吩丙氨酸、3-羟基亮氨酸、4-氮亮氨酸、5,5,5-三氟亮氨酸等)有抗性的大肠杆菌菌株(特公昭62-34397号和特开平8-70879号);通过WO96/06926中描述的遗传工程方法获得的大肠杆菌菌株;大肠杆菌H-9068(特开平8-70879号),等等。
可以通过增加涉及L-亮氨酸生物合成的1种以上的基因的表达来改进本发明的细菌。这些基因的实例可优选列举leuABCD操纵子中的基因,所述操纵子中的基因以经过突变的leuA基因为代表,该突变的leuA基因编码对L-亮氨酸反馈抑制具有抗性的异丙基苹果酸合酶(美国专利6,403,342)。此外,可以通过增强编码从细菌细胞分泌L-氨基酸的蛋白的1种以上的基因的表达来改进本发明的细菌。这类基因的实例包括b2682和b2683基因(ygaZH基因)(EP 1239041 A2)。
L-组氨酸生产菌
L-组氨酸生产菌或用于衍生L-组氨酸生产菌的亲本株的实例包括、但不限于下述属于埃希氏菌属的菌株,如大肠杆菌24菌株(VKPM B-5945,RU2003677);大肠杆菌80菌株(VKPM B-7270,RU2119536);大肠杆菌NRRL B-12116-B12121(美国专利4,388,405);大肠杆菌H-9342(FERMBP-6675)和H-9343(FERM BP-6676)(美国专利6,344,347);大肠杆菌H-9341(FERM BP-6674)(EP1085087);大肠杆菌AI80/pFM201(美国专利6,258,554)等等。
L-组氨酸生产菌或用于衍生L-组氨酸生产菌的亲本株的实例还包括如下菌株,所述菌株中编码L-组氨酸生物合成系统酶的1种以上的基因的表达增强。相关基因的实例包括ATP磷酸核糖基转移酶基因(hisG)、磷酸核糖基AMP环化水解酶基因(hisI)、磷酸核糖基-ATP焦磷酸水解酶基因(hisI)、磷酸核糖基亚氨甲基-5-氨基咪唑氨甲酰核糖核苷酸异构酶基因(phosphoribosylformimino-5-aminoimidazole carboxamide ribotide isomerase)(hisA)、酰胺转移酶基因(hisH)、组氨醇磷酸氨基转移酶基因(hisC)、组氨醇磷酸酶基因(hisB)、组氨醇脱氢酶基因(hisD)等。
已知hisG和hisBHAFI编码的L-组氨酸生物合成系统酶被L-组氨酸所抑制,因此还可以通过向ATP磷酸核糖基转移酶基因(hisG)中引入诱导对反馈抑制有抗性的突变来有效地增加产生L-组氨酸的能力(俄罗斯专利2003677和2119536)。
具有L-组氨酸生产能力的菌株具体实例包括大肠杆菌FERM-P 5038和5048,其已经导入了携带编码L-组氨酸生物合成系统酶的DNA的载体(特开昭56-005099号),导入了用于氨基酸输送的基因的大肠杆菌菌株(EP1016710A),赋予了磺胺胍、DL-1,2,4-三唑-3-丙氨酸和链霉素抗性的大肠杆菌80菌株(VKPM B-7270,俄罗斯专利2119536号),等等。
L-谷氨酸生产菌
L-谷氨酸生产菌或用于衍生L-谷氨酸生产菌的亲本株的实例包括但不限于大肠杆菌VL334thrC+(EP1172433)等属于埃希氏菌属的菌株。大肠杆菌VL334(VKPM B-1641)是L-异亮氨酸和L-苏氨酸营养缺陷型,在thrC和ilvA基因中有突变(美国专利4,278,765)。利用可在野生型大肠杆菌菌株K12(VKPM B-7)中增殖的噬菌体P1,通过常规转导转移了thrC基因的野生型等位基因。结果,获得了L-异亮氨酸营养缺陷型的L-谷氨酸生产菌VL334thrC+(VKPM B-8961)。
L-谷氨酸生产菌或用于衍生L-谷氨酸生产菌的亲本株的实例包括、但不限于编码L-谷氨酸生物合成系统酶的1种以上的基因的表达增加的菌株。作为相关基因的实例,可列举谷氨酸脱氢酶(gdhA)、谷氨酰胺合成酶(glnA)、谷氨酸合成酶(gltAB)、异柠檬酸脱氢酶(icdA)、顺乌头酸水合酶(acnA,acnB)、柠檬酸合酶(citrate synthase;gltA)、磷酸烯醇丙酮酸羧化酶(ppc)、丙酮酸脱氢酶(aceEF,lpdA)、丙酮酸激酶(pykA,pykF)、磷酸烯醇丙酮酸合酶(ppsA)、烯醇化酶(eno)、磷酸甘油酸变位酶(pgmA,pgmI)、磷酸甘油酸激酶(pgk)、甘油醛-3-磷酸脱氢酶(gapA)、丙糖磷酸异构酶(tpiA)、果糖二磷酸醛缩酶(fbp)、磷酸果糖激酶(pfkA,pfkB)和葡糖磷酸异构酶(pgi)等。
经修饰而增加了柠檬酸合成酶(citrate synthetase)基因、磷酸烯醇丙酮酸羧化酶基因和/或谷氨酸脱氢酶基因的表达增强的菌株的实例包括在EP1078989A、EP955368A和EP952221A中公开的那些菌株。
L-谷氨酸生产菌或用于衍生L-谷氨酸生产菌的亲本株的实例还包括下述酶的活性减少或缺损的菌株,所述酶催化从L-谷氨酸生物合成途径中分支的L-谷氨酸以外的化合物的合成。这类酶的实例包括异柠檬酸裂合酶(aceA),α-酮戊二酸脱氢酶(sucA),磷酸转乙酰酶(pta),乙酸激酶(ack),乙酰羟酸合酶(acetohydorxy acid synthase)(ilvG),乙酰乳酸合酶(ilvI),甲酸乙酰转移酶(pfl),乳酸脱氢酶(ldh),谷氨酸脱羧酶(gadAB)等。在美国专利5,378,616和5,573,945中描述了α-酮戊二酸脱氢酶活性缺失,或α-酮戊二酸脱氢酶活性降低的属于埃希氏菌属的细菌及它们的获得方法。
具体地,这些菌株可列举如下:
大肠杆菌W3110sucA::Kmr
大肠杆菌AJ12624(FERM BP-3853)
大肠杆菌AJ12628(FERM BP-3854)
大肠杆菌AJ12949(FERM BP-4881)。
大肠杆菌W3110sucA::Kmr是通过破坏大肠杆菌W3110的α-酮戊二酸脱氢酶基因(在下文也称为“sucA基因”)而获得的菌株。该菌株的α-酮戊二酸脱氢酶完全缺失。
L-谷氨酸生产菌的其它实例包括属于埃希氏菌属并且具有对天冬氨酸抗代谢物的抗性的细菌。这些菌株也可以是α-酮戊二酸脱氢酶缺陷的菌株,包括例如大肠杆菌AJ13199(FERM BP-5807)(美国专利5,908,768);进一步降低了L-谷氨酸分解能力的FERM P-12379(美国专利5,393,671);AJ13138(FERM BP-5565)(美国专利6,110,714),等等。
L-谷氨酸生产菌的实例还包括α-酮戊二酸脱氢酶活性缺陷或α-酮戊二酸脱氢酶活性降低的属于泛菌属的细菌,并且可以如上所述获得。这些菌株包括Pantoea ananatis AJ13356(美国专利6,331,419)。将Pantoea ananatisAJ13356于1998年2月19日保藏在工业技术院生命工学工业技术研究所(现在称作独立行政法人产业技术综合研究所特许微生物保藏中心,邮政信箱305-8566日本国茨城县筑波市东1丁目1番地1中央第6),保藏号为FERM P-16645。其后按照布达佩斯条约的规定,在1999年1月11日将其转为国际保藏,并得到保藏号FERM BP-6616。Pantoea ananatis AJ13356由于αKGDH-E1亚基基因(sucA)的破坏而缺失了α-酮戊二酸脱氢酶活性。当将上述菌株分离时将其鉴定为成团肠杆菌,并且作为成团肠杆菌AJ13356保藏。但是,近来基于对16S rRNA的核苷酸测序等,将其重新归类为Pantoeaananatis。尽管AJ13356作为成团肠杆菌保藏在前述保藏单位,就本说明书而言,将它们描述为Pantoea ananatis。
L-苯丙氨酸生产菌
L-苯丙氨酸生产菌或用于衍生L-苯丙氨酸生产菌的亲本株的例子包括、但不限于下述属于埃希氏菌属的菌株,如大肠杆菌AJ12739(tyrA::Tn10,tyrR)(VKPM B-8197);具有突变型pheA34基因的大肠杆菌HW1089(ATCC55371)(美国专利5,354,672);大肠杆菌MWEC101-b(KR8903681);大肠杆菌NRRL B-12141,NRRL B-12145,NRRL B-12146和NRRL B-12147(美国专利4,407,952)等。而且,也可以使用大肠杆菌K-12[W3110(tyrA)/pPHAB](FERM BP-3566),大肠杆菌K-12[W3110(tyrA)/pPHAD](FERM BP-12659),大肠杆菌K-12[W3110(tyrA)/pPHATerm](FERM BP-12662)和命名为AJ12604的大肠杆菌K-12[W3110(tyrA)/pBR-aroG4,pACMAB](FERMBP-3579)作为亲本菌株(EP 488424 B1)。此外,还可使用增加了yedA基因或yddG基因所编码蛋白的活性的属于埃希氏菌属的L-苯丙氨酸生产菌(美国专利申请2003/0148473 A1和2003/0157667 A1)。
L-色氨酸生产菌
L-色氨酸生产菌或用于衍生L-色氨酸生产菌的亲本株的实例包括、但不限于下述属于埃希氏菌属的菌株,如可以使用由突变的trpS基因编码的色氨酰-tRNA合成酶缺陷的大肠杆菌JP4735/pMU3028(DSM10122)和JP6015/pMU91(DSM10123)(美国专利5,756,345);大肠杆菌SV164(pGH5),其具有编码不受丝氨酸反馈抑制的磷酸甘油酸脱氢酶的serA等位基因和编码不受色氨酸反馈抑制的邻氨基苯甲酸合酶(anthranilate synthase)的trpE等位基因(美国专利6,180,373);色氨酸酶缺陷的大肠杆菌AGX17(pGX44)(NRRL B-12263)和AGX6(pGX50)aroP(NRRL B-12264)(美国专利4,371,614);产生磷酸烯醇丙酮酸的能力增加的大肠杆菌AGX17/pGX50,pACKG4-pps(WO9708333,美国专利6,319,696)等等。此外,还可以使用由yedA基因或yddG基因编码的蛋白活性增加的产生L-色氨酸的埃希氏菌属细菌(美国专利申请2003/0148473 A1和2003/0157667 A1)。
L-色氨酸生产菌或用于衍生L-色氨酸生产菌的亲本株的实例还包括增加了选自邻氨基苯甲酸合酶(trpE)、磷酸甘油酸脱氢酶(serA)和色氨酸合酶(trpAB)中的一种以上酶活性的菌株。邻氨基苯甲酸合酶和磷酸甘油酸脱氢酶同受L-色氨酸和L-丝氨酸的反馈抑制,因此可以向这些酶中引入导致反馈抑制解除的突变。具有此类突变的菌株的具体实例包括具有脱敏型邻氨基苯甲酸合酶的大肠杆菌SV164菌株和通过将质粒pGH5转入大肠杆菌SV164获得的菌株(WO 94/08031),所述质粒pGH5包含编码解除了反馈抑制的磷酸甘油酸脱氢酶的突变serA基因。
L-色氨酸生产菌或用于衍生L-色氨酸生产菌的亲本株的实例还包括导入了如下所述的色氨酸操纵子的菌株,所述色氨酸操纵子包含编码抑制解除型邻氨基苯甲酸合酶的基因(特开昭57-71397号,特开昭62-244382号,美国专利4,371,614)。此外,可以通过增加色氨酸操纵子(trpBA)中编码色氨酸合酶的基因的表达来赋予产生L-色氨酸的能力。色氨酸合酶由α和β亚基组成,α和β亚基分别由trpA和trpB编码。另外,可以通过增强异柠檬酸裂合酶-苹果酸合酶操纵子的表达来改进产生L-色氨酸的能力(WO2005/103275)。
L-脯氨酸生产菌
L-脯氨酸生产菌或用于衍生L-脯氨酸生产菌的亲本株实例包括、但不限于下述属于埃希氏菌属的菌株,如ilvA基因缺陷并且能够产生L-脯氨酸的大肠杆菌702ilvA(VKPM B-8012)(EP 1172433)等。
可以通过增加涉及L-脯氨酸生物合成的1种以上的基因的表达来改进本发明的细菌。用于L-脯氨酸生产菌的优选基因的实例包括编码L-脯氨酸反馈抑制解除的谷氨酸激酶的proB基因(德国专利第3127361号)。此外,可以通过增加1种以上编码从细菌细胞分泌L-氨基酸的蛋白的基因的表达来改进本发明的细菌。这些基因包括b2682和b2683基因(ygaZH基因)(EP1239041 A2)。
具有L-脯氨酸生产能力的属于埃希氏菌属的细菌的实例包括下列大肠杆菌菌株:NRRL B-12403和NRRL B-12404(英国专利第2075056号),VKPM B-8012(俄罗斯专利申请2000124295),德国专利第3127361中描述的质粒突变体,Bloom F.R.等描述的质粒突变体(The 15th Miami wintersymposium(第15届迈阿密冬季研讨会),1983,第34页)等等。
L-精氨酸生产菌
L-精氨酸生产菌或用于衍生L-精氨酸生产菌的亲本株包括、但不限于下述属于埃希氏菌属的菌株,如大肠杆菌237菌株(VKPM B-7925)(美国专利申请2002/058315 A1)及其带有突变的N-乙酰谷氨酸合酶(N-acetylglutamate synthase)的衍生菌株(俄罗斯专利申请No.2001112869),大肠杆菌382菌株(VKPM B-7926)(EP1170358A1),引入了编码N-乙酰谷氨酸合成酶(N-acetylglutamate synthetase)的argA基因的产生精氨酸的菌株(EP1170361A1),等等。
L-精氨酸生产菌或用于衍生L-精氨酸生产菌的亲本株包括、但不限于编码L-精氨酸生物合成酶的1种以上的基因的表达增加的菌株。相关基因的实例包括N-乙酰谷氨酰磷酸还原酶基因(argC)、鸟氨酸乙酰转移酶基因(argJ)、N-乙酰谷氨酸激酶基因(argB)、乙酰鸟氨酸转氨酶基因(argD)、鸟氨酸氨甲酰基转移酶基因(argF)、精氨琥珀酸合成酶基因(argG)、精氨琥珀酸裂合酶基因(argH)和氨甲酰基磷酸合成酶基因(carAB)。
L-缬氨酸生产菌
L-缬氨酸生产菌或用于衍生L-缬氨酸生产菌的亲本株实例包括、但不限于经修饰以过表达ilvGMEDA操纵子的菌株(美国专利5,998,178)。理想的是将衰减所需的ilvGMEDA操纵子的区域移除以使操纵子的表达不会被产生的L-缬氨酸所削弱。此外,理想的是将操纵子中的ilvA基因破坏从而降低苏氨酸脱氨酶的活性。
L-缬氨酸生产菌或用于衍生L-缬氨酸生产菌的亲本株的实例还包括具有氨酰t-RNA合成酶突变的突变株(美国专利5,658,766)。例如,可以使用大肠杆菌VL1970,该菌株在编码异亮氨酸tRNA合成酶的ileS基因中具有突变。已将大肠杆菌VL1970在1988年6月24日保藏在俄罗斯国立工业微生物保藏中心(VKPM)(Russia,113545Moscow,1 Dorozhny Proezd.),保藏号为VKPM B-4411。
此外,还可以使用生长需要硫辛酸(lipoic acid)的突变株和/或缺失H+-ATP酶的突变株(WO96/06926)。
L-异亮氨酸生产菌
L-异亮氨酸生产菌或用于衍生L-异亮氨酸生产菌的亲本株的实例包括、但不限于对6-二甲基氨基嘌呤有抗性的突变株(特开平5-304969号),对异亮氨酸类似物如硫代异亮氨酸(thiaisoleucine)和异亮氨酸氧肟酸具有抗性的突变株,以及对DL-乙硫氨酸(DL-ethionine)和/或精氨酸氧肟酸具有抗性的突变株(特开平5-130882号)。此外,还可以使用以编码涉及L-异亮氨酸生物合成的蛋白(如苏氨酸脱氨酶和乙酰羟酸合酶(acetohydroxatesynthase))的基因转化的重组菌株(特开平2-458号,FR 0356739,和美国专利第5,998,178号)作为亲本菌株。
实施例
[实施例1]果糖6磷酸醛缩酶和甘油脱氢酶和二羟基丙酮激酶活性增强的L-赖氨酸生产菌的构建
<1-1>dak1基因表达用质粒的构建
酿酒酵母染色体的全碱基序列已经阐明(Science 25(1996))。基于该文献报告的dak1基因的碱基序列,使用SEQ ID NO:14的合成寡核苷酸作为5’侧引物,使用SEQ ID NO:15所述的合成寡核苷酸作为3’侧引物,以酿酒酵母JCM7255株的染色体DNA为模板进行了PCR。将纯化后的PCR产物与用HindIII、SalI消化的载体pMW119(TAKARABIO公司制造)相连接,构建了dak1表达用质粒pMW-dak1。而且,JCM7255是保藏在日本琦玉县和光市广泽2-1(独立行政法人)理化学研究所“微生物系统保藏设施”的菌株。
<1-2>甘油脱氢酶活性提高株的构建
构建了经过修饰而具有SEQ ID NO:11所示结构的WC196ΔcadAΔldcC株。为了构架具有该结构的菌株,使用了SEQ ID NO:9所示的序列(PCR产物)。在SEQ ID NO:9所示的序列中,碱基序号第1位~第172位是λ噬菌体的attR序列,碱基序号第324位~第983位是氯霉素抗性基因(cat),碱基序号第1540位~第1653位是λ噬菌体的attL序列,碱基序号第1654位~第1733位是tacM启动子。
而且,tacM启动子(SEQ ID NO:10)可通过将tac启动子(Gene 25(2-3),167-178(1983))的-35区的TTGACA序列替换为TTCACA来构建。SEQ IDNO:9的序列可参考pMW118-attL-Cm-attR(WO2005/010175)来构建。
以SEQ ID NO:9的序列为模板,通过使用SEQ ID NO:12和SEQ IDNO:13所示引物的PCR进行扩增,将该扩增产物通过λ-RED法(WO2005/010175)插入到WC196ΔcadAΔldcC株(参照国际公布第WO2006/038695号小册子)的染色体上,获得了gldA上游的启动子序列被替换的菌株。这样一来,得到了甘油脱氢酶活性提高的菌株:WC196ΔcadAΔldcCPtacMgldA::Cm株。
<1-3>果糖6磷酸醛缩酶和甘油脱氢酶活性增强的L-赖氨酸生产菌的构建
构建了经过修饰而具有SEQ ID NO:92所示结构的WC196ΔcadAΔldcC株。为了构建具有该结构的菌株,使用了SEQ ID NO:9所示的序列(PCR产物)。在SEQ ID NO:9所示的序列中,碱基序号第1位~第172位为λ噬菌体的attR序列,碱基序号第324位~第983位为氯霉素抗性基因(cat),碱基序号第1540位~第1653位为λ噬菌体的attL序列,碱基序号第1654位~第1733位为tacM启动子。
而且,tacM启动子(SEQ ID NO:10)可通过将tac启动子(Gene 25(2-3),167-178(1983))的-35区的TTGACA序列替换成TTCACA来构建。SEQ IDNO:9的序列可参考pMW118-attL-Cm-attR(WO2005/010175)来构建。
以SEQ ID NO:9的序列为模板,通过使用SEQ ID NO:93和SEQ IDNO:94所示引物的PCR进行了扩增,将该扩增产物采用λ-RED法(WO2005/010175)插入WC196ΔcadAΔldcC株(参照国际公布第WO2006/038695号小册子)的染色体上,得到了fsaB-gldA操纵子上游的启动子序列被替换的菌株。由此,得到了果糖6磷酸醛缩酶和甘油脱氢酶活性提高的菌株WC196ΔcadAΔldcCPtacM fsaB-gldA::Cm株。
<1-4>果糖6磷酸醛缩酶和甘油脱氢酶和二羟基丙酮激酶活性增强的L-赖氨酸生产菌的构建
对于WC196ΔcadAΔldcC株(参照国际公布第WO2006/038695号小册子)、WC196ΔcadAΔldcCPtacMgldA::Cm株和WC196ΔcadAΔldcCPtacMfsaB-gldA::Cm株,使用携带dapA、dapB和LysC基因的Lys生产用质粒pCABD2(国际公布第WO01/53459号小册子)按常规方法进行转化,得到了WC196ΔcadAΔldcC/pCABD2株、WC196ΔcadAΔ1dcCPtacMgldA::Cm/pCABD2株、WC196ΔcadAΔldcCPtacMfsaB-gldA::Cm/pCABD2株。而且,对于WC196ΔcadAΔldcC/pCABD2株、WC196ΔcadAΔldcCPtacMgldA::Cm/pCABD2株、WC196ΔcadAΔ1dcCPtacMfsaB-gldA::Cm/pCABD2株,用dak1表达用质粒pMW-dak1按常规方法进行转化,得到了WC196ΔcadAΔldcC/pCABD2,pMW-dak1株、WC196ΔcadAΔldcCPtacMgldA::Cm/pCABD2,pMW-dak1株、和WC196ΔcadAΔ1dcCPtacM fsaB-gldA::Cm/pCABD2,pMW-dak1株。
将这些菌株在含20mg/L的链霉素、或20mg/L的链霉素和50mg/L的氨苄青霉素的L培养基中于37℃培养至终然后,加入与培养液等量的40%甘油溶液并进行搅拌,然后以适当的量分装,于-80℃保存。将其称为甘油保存液。
[实施例2]果糖6磷酸醛缩酶和甘油脱氢酶和二羟基丙酮激酶活性增强的L-赖氨酸生产菌的评价
融化上述甘油保存液,各取100μL均匀涂布在含20mg/L的链霉素、或20mg/L的链霉素和50mg/L的氨苄青霉素的L平板上,37℃培养24小时。将所得的平板上的菌体悬浮在1ml的生理食盐水中,用常数50除以将悬浮液稀释101倍时的吸光度600nm的值(n),计算出液量(V)(V=50/n),将该液量接种到500mL坂口培养瓶中的、含20mg/L的链霉素、或20mg/L的链霉素和50mg/L的氨苄青霉素的发酵培养基20mL中,用往复式振荡培养装置37℃培养48小时。培养后,采用公知方法(SAKURA精机BIOTECHANALYSERAS210)测定培养基中蓄积的赖氨酸的量。
发酵培养基的组成如下(单位g/L)。
甘油 40
(NH4)2SO4 24
K2HPO4 1.0
MgSO4·7H2O 1.0
FeSO4·7H2O 0.01
MnSO4·5H2O 0.01
酵母提取物2.0
加至1L
用KOH调整至pH7.0,115℃高压灭菌10分钟(但甘油和MgSO4·7H2O另行灭菌)
添加符合日本药典的(局方)CaCO3 30g/L(180℃干热灭菌2小时)。
作为抗生素,添加20mg/L的链霉素、或20mg/L的链霉素加50mg/L的氨苄青霉素。培养在温度37℃、搅拌115rpm的条件下进行48小时。
结果示于表5(OD表示吸光度600nm下的菌体量、Lys(g/L)表示培养瓶中蓄积的L-赖氨酸蓄积量、产率[产率](%)表示从基质到L-赖氨酸的产率)。就单独强化甘油脱氢酶的菌株、单独强化二羟基丙酮激酶的菌株、和强化果糖6磷酸醛缩酶和甘油脱氢酶的菌株而言,与非修饰株相比,在收率、生产性方面均无变化;而在同时强化甘油脱氢酶和以ATP为磷酸供体的二羟基丙酮激酶的WC196ΔcadAΔldcCPtacMgldA::Cm/pCABD2,pMW-dak1株中,与其它菌株相比较,蓄积了大量的L-赖氨酸。此外,就同时强化果糖6磷酸醛缩酶和甘油脱氢酶和以ATP为磷酸供体的二羟基丙酮激酶的WC196ΔcadAΔldcCPtacM fsaB-gldA::Cm/pCABD2,pMW-dak1株而言,蓄积了更大量的L-赖氨酸。
[表5]:果糖6磷酸醛缩酶(fsaB)和甘油脱氢酶(gldA)和二羟基丙酮激酶(dak1)活性增强株的L-赖氨酸蓄积
O.D. | Lys(g/L) | 产率(%) | |
WC196LC pCABD2 - | 16.7 | 14.7 | 36.8 |
WC 196LC pCABD2pM W-dak1 | 14.3 | 14.8 | 36.9 |
WC 196LCP tacM gldA pCABD2 - | 18.1 | 14.7 | 36.8 |
WC 196LCP tacM fsaB-gldA pCABD2 - | 18.5 | 14.3 | 35.8 |
WC 196LCP tacM gldA pCABD2pM W-dak1 | 15.3 | 15.3 | 38.1 |
WC 196LCP tacM fsaB-gldA pCABD2pM W-dak1 | 14.0 | 16.9 | 42.1 |
表中的菌株名中,“LC”为“ΔcadAΔldcC”的简记,省略了“::Cm”。
[实施例3]增强了甘油脱氢酶和二羟基丙酮激酶活性的L-苏氨酸生产菌的构建
<3-1>甘油脱氢酶活性提高株的构建
构建了经过修饰而具有SEQ ID NO:90和91所示结构的B5318株。为了构建具有该结构的菌株,使用了SEQ ID NO:88和89所示的序列(PCR产物)。在SEQ ID NO:88和89所示的序列中,碱基序号第1位~第72位为λ噬菌体的attR序列,碱基序号第324位~第983位为氯霉素抗性基因(cat),碱基序号第1540位~第1653位为λ噬菌体的attL序列,碱基序号第1654位~第1733位为tacM2和tacM3启动子。
而且,tacM2和tacM3启动子是可通过将tac启动子(Gene 25(2-3),167-178(1983))的-35区的TTGACA序列替换为TGTACA和TTGGCA来构建的组成型启动子(Molecular Biology 39(5)719-726(2005))。SEQ ID NO:88和89的序列可参考pMW118-attL-Cm-attR(WO2005/010175)来构建。
以SEQ ID NO:88和89的序列为模板,通过使用SEQ ID NO:12和SEQ ID NO:13所示引物的PCR进行扩增,将该扩增产物用λ-RED法(WO2005/010175)插入到B5318株(VKPM B-5318)的染色体上,得到了gldA上游的启动子序列被替换的菌株。由此,得到了甘油脱氢酶活性提高的菌株即B5318PtacM2gldA::Cm株和B5318PtacM3gldA::Cm株。
<1-3>增强了甘油脱氢酶和二羟基丙酮激酶活性的L-苏氨酸生产菌的构建
对于B5318PtacM2gldA::Cm株和B5318PtacM3gldA::Cm株,使用dak1表达用质粒pMW-dak1按常规方法进行转化,得到了B5318PtacM2gldA::Cm/pMW-dak1株和B5318PtacM3gldA::Cm/pMW-dak1株。
将这些菌株在含20mg/L的链霉素、或20mg/L的链霉素及50mg/L的氨苄青霉素的L培养基中37℃培养至终然后,加入与培养液等量的40%甘油溶液并进行搅拌,然后以适当量分装,于-80℃保存。将其称为甘油保存液。
[实施例4]强化了甘油脱氢酶和二羟基丙酮激酶活性的L-苏氨酸生产菌的评价
融化上述甘油保存液,各取100μL均匀涂布在含20mg/L的链霉素、或20mg/L的链霉素和50mg/L的氨苄青霉素的L平板上,37℃培养24小时。将所得的平板上的菌体悬浮在1ml的生理食盐水中,用常数50除以将悬浮液稀释101倍时的吸光度600nm的值(n),计算出液量(V)(V=50/n),将该液量接种到500mL带挡板三角培养瓶中的含20mg/L的链霉素、或含20mg/L链霉素及50mg/L的氨苄青霉素的发酵培养基20mL中,用旋转培养装置于40℃培养24小时。培养后,采用公知方法(日立液相色谱,ODS-2柱)测定培养基中蓄积的苏氨酸的量。
发酵培养基的组成如下(单位g/L)。
甘油 40
(NH4)2SO4 24
K2HPO4 1.0
MgSO4·7H2O 1.0
FeSO4·7H2O 0.01
MnSO4·5H2O 0.01
酵母提取物 2.0
加至1L
用KOH调整至pH7.0,115℃高压灭菌10分钟(但甘油和MgSO4·7H2O另行灭菌)
添加符合日本药典的(局方)CaCO3 30g/L(180℃干热灭菌2小时)。
作为抗生素,添加了20mg/L的链霉素、或20mg/L的链霉素及50mg/L的氨苄青霉素。培养在温度40℃、搅拌144rpm旋转培养的条件下进行24小时。
结果示于表6(OD表示吸光度600nm下的菌体量,Thr(g/L)表示培养瓶中蓄积的L-苏氨酸蓄积量,产率[产率](%)表示从基质到L-苏氨酸的产率)。与非修饰株相比,单独强化甘油脱氢酶的菌株在产率、生产性方面均无变化,而在同时强化甘油脱氢酶和以ATP为磷酸供体的二羟基丙酮激酶的B5318PtacM2gldA::Cm/pMW-dak1株和B5318PtacM3gldA::Cm/pMW-dak1株中,与其它菌株相比较,蓄积了大量的L-苏氨酸。
[表6]:甘油脱氢酶(gldA)和二羟基丙酮激酶(dak1)活性增强株的L-苏氨酸蓄积
OD600 | Thr(g/L) | 产率(%) | |
B 5318 - - | 22.5 | 12.5 | 30.9 |
B 5318 Ptac M2 gldA - | 21.5 | 11.9 | 29.4 |
B 5318 Ptac M2 gldA pMW-dak | 21.1 | 13.2 | 32.6 |
B 5318 Ptac M3 gldA - | 23.1 | 12.3 | 30.4 |
B 5318 Ptac M3 gldA pMW-dak | 22.3 | 13.3 | 32.9 |
表中的株名中,“pMW-dak”为“pMW-dak1”的简记,省略了“::Cm”。
〔序列表的说明〕
SEQ ID NO:1:大肠杆菌的gldA基因序列(1104bp)
SEQ ID NO:2:大肠杆菌的GldA氨基酸序列(367AA)
SEQ ID NO:3:酿酒酵母的dakA1基因序列(1755bp)
SEQ ID NO:4:酿酒酵母的DakA氨基酸序列(584AA)
SEQ ID NO:5:根瘤土壤杆菌的dhbK1基因序列(1695bp)
SEQ ID NO:6:根瘤土壤杆菌的Dhbk1氨基酸序列(564AA)
SEQ ID NO:7:弗氏柠檬酸杆菌的dhaK基因序列(1659bp)
SEQ ID NO:8:弗氏柠檬酸杆菌的DhaK氨基酸序列(552AA)
SEQ ID NO:9:attR-cat-attL-ptacM-SD-间隔序列(1740bp)
SEQ ID NO:10:tacM启动子(80bp)
SEQ ID NO:11:PtacMgldA::Cm序列
SEQ ID NO:12:atL-Ptac-gldA(gldA染色体上强化用PCR引物)
SEQ ID NO:13:atR-Ptac-fsaB1(gldA染色体上强化用PCR引物)
SEQ ID NO:14:pMW-dak1F(dakA克隆用引物)
SEQ ID NO:15:pMW-dak1R(dakA克隆用引物)
SEQ ID NO:16:大肠杆菌的glpF基因序列(846bp)
SEQ ID NO:17:大肠杆菌的GlpF氨基酸序列(281AA)
SEQ ID NO:18:大肠杆菌的tpiA基因序列(768bp)
SEQ ID NO:19:大肠杆菌的TpiA氨基酸序列(255AA)
SEQ ID NO:20:大肠杆菌的fbaA基因序列(1080bp)
SEQ ID NO:21:大肠杆菌的FbaA氨基酸序列(359AA)
SEQ ID NO:22:大肠杆菌的glpX基因序列(1011bp)
SEQ ID NO:23:大肠杆菌的GlpX氨基酸序列(336AA)
SEQ ID NO:24:大肠杆菌的glpK基因序列(1509bp)
SEQ ID NO:25:大肠杆菌的GlpK氨基酸序列(502AA)
SEQ ID NO:26:大肠杆菌的glpA基因序列(1629bp)
SEQ ID NO:27:大肠杆菌的GlpA氨基酸序列(542AA)
SEQ ID NO:28:大肠杆菌的glpB基因序列(1260bp)
SEQ ID NO:29:大肠杆菌的GlpB氨基酸序列(419AA)
SEQ ID NO:30:大肠杆菌的glpC基因序列(1191bp)
SEQ ID NO:31:大肠杆菌的GlpC氨基酸序列(396AA)
SEQ ID NO:32:大肠杆菌的glpD基因序列(1506bp)
SEQ ID NO:33:大肠杆菌的GlpD氨基酸序列(501AA)
SEQ ID NO:34:大肠杆菌的dhaK基因序列(1071bp)
SEQ ID NO:35:大肠杆菌的DhaK氨基酸序列(356AA)
SEQ ID NO:36:大肠杆菌的dhaL基因序列(633bp)
SEQ ID NO:37:大肠杆菌的DhaL氨基酸序列(210AA)
SEQ ID NO:38:大肠杆菌的dhaM基因序列(1419bp)
SEQ ID NO:39;大肠杆菌的DhaM氨基酸序列(472AA)
SEQ ID NO:40:粟酒裂殖酵母的二羟基丙酮激酶基因(1776bp)
SEQ ID NO:41:粟酒裂殖酵母的二羟基丙酮激酶(591AA)
SEQ ID NO:42:Pichia angusta的二羟基丙酮激酶基因(1830bp)
SEQ ID NO:43:Pichia angusta的二羟基丙酮激酶(609AA)
SEQ ID NO:44:巴斯德毕赤酵母(Pichia pastoris)的二羟基丙酮激酶基因(1827bp)
SEQ ID NO:45:巴斯德毕赤酵母的二羟基丙酮激酶(608AA)
SEQ ID NO:46:汉森德巴利酵母(Debaryomyces hansenii)的二羟基丙酮激酶基因(1824bp)
SEQ ID NO:47:汉森德巴利酵母的二羟基丙酮激酶(607AA)
SEQ ID NO:48:蟑螂埃希氏菌(Escherichia blattae)的二羟基丙酮激酶基因(1752bp)
SEQ ID NO:49:蟑螂埃希氏菌的二羟基丙酮激酶(583AA)
SEQ ID NO:50:肠杆菌属菌种(Enterobacter sp.)638的二羟基丙酮激酶基因(1647bp)
SEQ ID NO:51:肠杆菌属菌种638的二羟基丙酮激酶(548AA)
SEQ ID NO:52:嗜冷单胞菌属菌种(Psychromonas sp.)CNPT3的二羟基丙酮激酶基因(1695bp)
SEQ ID NO:53:嗜冷单胞菌属菌种CNPT3的二羟基丙酮激酶(564AA)
SEQ ID NO:54:Stappia aggregata的二羟基丙酮激酶基因(1647bp)
SEQ ID NO:55:Stappia aggregata的二羟基丙酮激酶(548AA)
SEQ ID NO:56:豌豆根瘤菌蚕豆生物变种(Rhizobium leguminosarum bv.viciae)3841的二羟基丙酮激酶基因(1641bp)
SEQ ID NO:57:豌豆根瘤菌蚕豆生物变种3841的二羟基丙酮激酶(546AA)
SEQ ID NO:58:黄色粘球菌(Myxococcus xanthus)DK 1622的二羟基丙酮激酶基因(1701bp)
SEQ ID NO:59:黄色粘球菌DK 1622的二羟基丙酮激酶(566AA)
SEQ ID NO:60:伯克霍尔德氏菌属菌种(Burkholderia sp.)383的二羟基丙酮激酶基因(1701bp)
SEQ ID NO:61:伯克霍尔德氏菌属菌种383的二羟基丙酮激酶(566AA)
SEQ ID NO:62:泰国伯克霍尔德氏菌(Burkholderia thailandensis)E264的二羟基丙酮激酶基因(1704bp)
SEQ ID NO:63:泰国伯克霍尔德氏菌E264的二羟基丙酮激酶(567AA)
SEQ ID NO:64:多食伯克霍尔德氏菌(Burkholderia multivorans)ATCC17616的二羟基丙酮激酶基因(1851bp)
SEQ ID NO:65:多食伯克霍尔德氏菌ATCC 17616的二羟基丙酮激酶(616AA)
SEQ ID NO:66:大肠杆菌的dhaR基因(1920bp)
SEQ ID NO:67:大肠杆菌的DhaR氨基酸序列(639AA)
SEQ ID NO:68:大肠杆菌的fsaA基因(663bp)
SEQ ID NO:69:大肠杆菌的FsaA氨基酸序列(220AA)
SEQ ID NO:70:大肠杆菌的fsaB基因(663bp)
SEQ ID NO:71:大肠杆菌的FsaB氨基酸序列(220AA)
SEQ ID NO:72:大肠杆菌的fbaB基因(1053bp)
SEQ ID NO:73:大肠杆菌的FbaB氨基酸序列(350AA)
SEQ ID NO:74:痢疾志贺氏菌(Shigella dysenteriae)Sd197的gldA基因(1143bp)
SEQ ID NO:75:痢疾志贺氏菌Sd197的GldA氨基酸序列(380AA)
SEQ ID NO:76:鼠伤寒沙门氏菌(Salmonella typhimurium)LT2的gldA基因(1104bp)
SEQ ID NO:77:鼠伤寒沙门氏菌LT2的GldA氨基酸序列(367AA)
SEQ ID NO:78:恶臭假单胞菌(Pseudomonas putida)的gldA基因(1098bp)
SEQ ID NO:79:恶臭假单胞菌的GldA氨基酸序列(365AA)
SEQ ID NO:80:凝结芽孢杆菌(Bacillus coagulans)36D1的gldA基因(1104bp)
SEQ ID NO:81;凝结芽孢杆菌36D1的GldA氨基酸序列(367AA)
SEQ ID NO:82;大肠杆菌的fbp基因(999bp)
SEQ ID NO:83:大肠杆菌的Fbp氨基酸序列(322AA)
SEQ ID NO:84:大肠杆菌的ybhA基因(819bp)
SEQ ID NO:85:大肠杆菌的YbhA氨基酸序列(272AA)
SEQ ID NO:86:大肠杆菌的ptsI基因(1782bp)
SEQ ID NO:87:大肠杆菌的PtsI氨基酸序列(575AA)
SEQ ID NO:88:attR-cat-attL-PtacM2-SD-间隔序列
SEQ ID NO:89:attR-cat-attL-PtacM3-SD-间隔序列
SEQ ID NO:90:PtacM2gldA::Cm序列
SEQ ID NO:91:PtacM3gldA::Cm序列
SEQ ID NO:92:PtacMfsaB-gldA::Cm序列
SEQ ID NO:93:atL-Ptac-fsaB(fsaB+gldA染色体上强化用PCR引物)
SEQ ID NO:94:atR-Ptac-fsaB(fsaB+gldA染色体上强化用PCR引物)
工业实用性
通过使用本发明的微生物,可以高效地从甘油发酵生产L-氨基酸。
序列表
<110>味之素株式会社(Ajinomoto Co.,Inc.)
<120>L-氨基酸的生产方法
<130>C876-C7306
<150>JP 2007-041724
<151>2007-02-22
<160>94
<170>PatentIn version 3.1
<210>1
<211>1104
<212>DNA
<213>大肠杆菌(Escherichia coli)
<220>
<221>CDS
<222>(1)..(1104)
<223>
<400>1
atg gac cgc att att caa tca ccg ggt aaa tac atc cag ggc gct gat 48
Met Asp Arg Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp
1 5 10 15
gtg att aat cgt ctg ggc gaa tac ctg aag ccg ctg gca gaa cgc tgg 96
Val Ile Asn Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Arg Trp
20 25 30
tta gtg gtg ggt gac aaa ttt gtt tta ggt ttt gct caa tcc act gtc 144
Leu Val Val Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val
35 40 45
gag aaa agc ttt aaa gat gct gga ctg gta gta gaa att gcg ccg ttt 192
Glu Lys Ser Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe
50 55 60
ggc ggt gaa tgt tcg caa aat gag atc gac cgt ctg cgt ggc atc gcg 240
Gly Gly Glu Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala
65 70 75 80
gag act gcg cag tgt ggc gca att ctc ggt atc ggt ggc gga aaa acc 288
Glu Thr Ala Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr
85 90 95
ctc gat act gcc aaa gca ctg gca cat ttc atg ggt gtt ccg gta gcg 336
Leu Asp Thr Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala
100 105 110
atc gca ccg act atc gcc tct acc gat gca ccg tgc agc gca ttg tct 384
Ile Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser
115 120 125
gtt atc tac acc gat gag ggt gag ttt gac cgc tat ctg ctg ttg cca 432
Val Ile Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro
130 135 140
aat aac ccg aat atg gtc att gtc gac acc aaa atc gtc gct ggc gca 480
Asn Asn Pro Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Ala
145 150 155 160
cct gca cgt ctg tta gcg gcg ggt atc ggc gat gcg ctg gca acc tgg 528
Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp
165 170 175
ttt gaa gcg cgt gcc tgc tct cgt agc ggc gcg acc acc atg gcg ggc 576
Phe Glu Ala Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly
180 185 190
ggc aag tgc acc cag gct gcg ctg gca ctg gct gaa ctg tgc tac aac 624
Gly Lys Cys Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn
195 200 205
acc ctg ctg gaa gaa ggc gaa aaa gcg atg ctt gct gcc gaa cag cat 672
Thr Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His
210 215 220
gta gtg act ccg gcg ctg gag cgc gtg att gaa gcg aac acc tat ttg 720
Val Val Thr Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu
225 230 235 240
agc ggt gtt ggt ttt gaa agt ggt ggt ctg gct gcg gcg cac gca gtg 768
Ser Gly Val Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val
245 250 255
cat aac ggc ctg acc gct atc ccg gac gcg cat cac tat tat cac ggt 816
His Asn Gly Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly
260 265 270
gaa aaa gtg gca ttc ggt acg ctg acg cag ctg gtt ctg gaa aat gcg 864
Glu Lys Val Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala
275 280 285
ccg gtg gag gaa atc gaa acc gta gct gcc ctt agc cat gcg gta ggt 912
Pro Val Glu Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly
290 295 300
ttg cca ata act ctc gct caa ctg gat att aaa gaa gat gtc ccg gcg 960
Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Lys Glu Asp Val Pro Ala
305 310 315 320
aaa atg cga att gtg gca gaa gcg gca tgt gca gaa ggt gaa acc att 1008
Lys Met Arg Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile
325 330 335
cac aac atg cct ggc ggc gcg acg cca gat cag gtt tac gcc gct ctg 1056
His Asn Met Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu
340 345 350
ctg gta gcc gac cag tac ggt cag cgt ttc ctg caa gag tgg gaa taa 1104
Leu Val Ala Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu
355 360 365
<210>2
<211>367
<212>PRT
<213>大肠杆菌
<400>2
Met Asp Arg Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp
1 5 10 15
Val Ile Asn Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Arg Trp
20 25 30
Leu Val Val Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val
35 40 45
Glu Lys Ser Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe
50 55 60
Gly Gly Glu Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala
65 70 75 80
Glu Thr Ala Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr
85 90 95
Leu Asp Thr Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala
100 105 110
Ile Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser
115 120 125
Val Ile Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro
130 135 140
Asn Asn Pro Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Ala
145 150 155 160
Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp
165 170 175
Phe Glu Ala Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly
180 185 190
Gly Lys Cys Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn
195 200 205
Thr Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His
210 215 220
Val Val Thr Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu
225 230 235 240
Ser Gly Val Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val
245 250 255
His Asn Gly Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly
260 265 270
Glu Lys Val Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala
275 280 285
Pro Val Glu Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly
290 295 300
Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Lys Glu Asp Val Pro Ala
305 310 315 320
Lys Met Arg Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile
325 330 335
His Asn Met Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu
340 345 350
Leu Val Ala Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu
355 360 365
<210>3
<211>1755
<212>DNA
<213>酿酒酵母(Saccharomyces cerevisiae)
<220>
<221>CDS
<222>(1)..(1755)
<223>
<400>3
atg tcc gct aaa tcg ttt gaa gtc aca gat cca gtc aat tca agt ctc 48
Met Ser Ala Lys Ser Phe Glu Val Thr Asp Pro Val Asn Ser Ser Leu
1 5 10 15
aaa ggg ttt gcc ctt gct aac ccc tcc att acg ctg gtc cct gaa gaa 96
Lys Gly Phe Ala Leu Ala Asn Pro Ser Ile Thr Leu Val Pro Glu Glu
20 25 30
aaa att ctc ttc aga aag acc gat tcc gac aag atc gca tta att tct 144
Lys Ile Leu Phe Arg Lys Thr Asp Ser Asp Lys Ile Ala Leu Ile Ser
35 40 45
ggt ggt ggt agt gga cat gaa cct aca cac gcc ggt ttc att ggt aag 192
Gly Gly Gly Ser Gly His Glu Pro Thr His Ala Gly Phe Ile Gly Lys
50 55 60
ggt atg ttg agt ggc gcc gtg gtt ggc gaa att ttt gca tcc cct tca 240
Gly Met Leu Ser Gly Ala Val Val Gly Glu Ile Phe Ala Ser Pro Ser
65 70 75 80
aca aaa cag att tta aat gca atc cgt tta gtc aat gaa aat gcg tct 288
Thr Lys Gln Ile Leu Asn Ala Ile Arg Leu Val Asn Glu Asn Ala Ser
85 90 95
ggc gtt tta ttg att gtg aag aac tac aca ggt gat gtt ttg cat ttt 336
Gly Val Leu Leu Ile Val Lys Asn Tyr Thr Gly Asp Val Leu His Phe
100 105 110
ggt ctg tcc gct gag aga gca aga gcc ttg ggt att aac tgc cgc gtt 384
Gly Leu Ser Ala Glu Arg Ala Arg Ala Leu Gly Ile Asn Cys Arg Val
115 120 125
gct gtc ata ggt gat gat gtt gca gtt ggc aga gaa aag ggt ggt atg 432
Ala Val Ile Gly Asp Asp Val Ala Val Gly Arg Glu Lys Gly Gly Met
130 135 140
gtt ggt aga aga gca ttg gca ggt acc gtt ttg gtt cat aag att gta 480
Val Gly Arg Arg Ala Leu Ala Gly Thr Val Leu Val His Lys Ile Val
145 150 155 160
ggt gcc ttc gca gaa gaa tat tct agt aag tat ggc tta gac ggt aca 528
Gly Ala Phe Ala Glu Glu Tyr Ser Ser Lys Tyr Gly Leu Asp Gly Thr
165 170 175
gct aaa gtg gct aaa att atc aac gac aat ttg gtg acc att gga tct 576
Ala Lys Val Ala Lys Ile Ile Asn Asp Asn Leu Val Thr Ile Gly Ser
180 185 190
tct tta gac cat tgt aaa gtt cct ggc agg aaa ttc gaa agt gaa tta 624
Ser Leu Asp His Cys Lys Val Pro Gly Arg Lys Phe Glu Ser Glu Leu
195 200 205
aac gaa aaa caa atg gaa ttg ggt atg ggt att cat aac gaa cct ggt 672
Asn Glu Lys Gln Met Glu Leu Gly Met Gly Ile His Asn Glu Pro Gly
210 215 220
gtg aaa gtt tta gac cct att cct tct acc gaa gac ttg atc tcc aag 720
Val Lys Val Leu Asp Pro Ile Pro Ser Thr Glu Asp Leu Ile Ser Lys
225 230 235 240
tat atg cta cca aaa cta ttg gat cca aac gat aag gat aga gct ttt 768
Tyr Met Leu Pro Lys Leu Leu Asp Pro Asn Asp Lys Asp Arg Ala Phe
245 250 255
gta aag ttt gat gaa gat gat gaa gtt gtc ttg tta gtt aac aat ctc 816
Val Lys Phe Asp Glu Asp Asp Glu Val Val Leu Leu Val Asn Asn Leu
260 265 270
ggc ggt gtt tct aat ttt gtt att agt tct atc act tcc aaa act acg 864
Gly Gly Val Ser Asn Phe Val Ile Ser Ser Ile Thr Ser Lys Thr Thr
275 280 285
gat ttc tta aag gaa aat tac aac ata acc ccg gtt caa aca att gct 912
Asp Phe Leu Lys Glu Asn Tyr Asn Ile Thr Pro Val Gln Thr Ile Ala
290 295 300
ggc aca ttg atg acc tcc ttc aat ggt aat ggg ttc agt atc aca tta 960
Gly Thr Leu Met Thr Ser Phe Asn Gly Asn Gly Phe Ser Ile Thr Leu
305 310 315 320
cta aac gcc act aag gct aca aag gct ttg caa tct gat ttt gag gag 1008
Leu Asn Ala Thr Lys Ala Thr Lys Ala Leu Gln Ser Asp Phe Glu Glu
325 330 335
atc aaa tca gta cta gac ttg ttg aac gca ttt acg aac gca ccg ggc 1056
Ile Lys Ser Val Leu Asp Leu Leu Asn Ala Phe Thr Asn Ala Pro Gly
340 345 350
tgg cca att gca gat ttt gaa aag act tct gcc cca tct gtt aac gat 1104
Trp Pro Ile Ala Asp Phe Glu Lys Thr Ser Ala Pro Ser Val Asn Asp
355 360 365
gac ttg tta cat aat gaa gta aca gca aag gcc gtc ggt acc tat gac 1152
Asp Leu Leu His Asn Glu Val Thr Ala Lys Ala Val Gly Thr Tyr Asp
370 375 380
ttt gac aag ttt gct gag tgg atg aag agt ggt gct gaa caa gtt atc 1200
Phe Asp Lys Phe Ala Glu Trp Met Lys Ser Gly Ala Glu Gln Val Ile
385 390 395 400
aag agc gaa ccg cac att acg gaa cta gac aat caa gtt ggt gat ggt 1248
Lys Ser Glu Pro His Ile Thr Glu Leu Asp Asn Gln Val Gly Asp Gly
405 410 415
gat tgt ggt tac act tta gtg gca gga gtt aaa ggc atc acc gaa aac 1296
Asp Cys Gly Tyr Thr Leu Val Ala Gly Val Lys Gly Ile Thr Glu Asn
420 425 430
ctt gac aag ctg tcg aag gac tca tta tct cag gcg gtt gcc caa att 1344
Leu Asp Lys Leu Ser Lys Asp Ser Leu Ser Gln Ala Val Ala Gln Ile
435 440 445
tca gat ttc att gaa ggc tca atg gga ggt act tct ggt ggt tta tat 1392
Ser Asp Phe Ile Glu Gly Ser Met Gly Gly Thr Ser Gly Gly Leu Tyr
450 455 460
tct att ctt ttg tcg ggt ttt tca cac gga tta att cag gtt tgt aaa 1440
Ser Ile Leu Leu Ser Gly Phe Ser His Gly Leu Ile Gln Val Cys Lys
465 470 475 480
tca aag gat gaa ccc gtc act aag gaa att gtg gct aag tca ctc gga 1488
Ser Lys Asp Glu Pro Val Thr Lys Glu Ile Val Ala Lys Ser Leu Gly
485 490 495
att gca ttg gat act tta tac aaa tat aca aag gca agg aag gga tca 1536
Ile Ala Leu Asp Thr Leu Tyr Lys Tyr Thr Lys Ala Arg Lys Gly Ser
500 505 510
tcc acc atg att gat gct tta gaa cca ttc gtt aaa gaa ttt act gca 1584
Ser Thr Met Ile Asp Ala Leu Glu Pro Phe Val Lys Glu Phe Thr Ala
515 520 525
tct aag gat ttc aat aag gcg gta aaa gct gca gag gaa ggt gct aaa 1632
Ser Lys Asp Phe Asn Lys Ala Val Lys Ala Ala Glu Glu Gly Ala Lys
530 535 540
tcc act gct aca ttc gag gcc aaa ttt ggc aga gct tcg tat gtc ggc 1680
Ser Thr Ala Thr Phe Glu Ala Lys Phe Gly Arg Ala Ser Tyr Val Gly
545 550 555 560
gat tca tct caa gta gaa gat cct ggt gca gta ggc cta tgt gag ttt 1728
Asp Ser Ser Gln Val Glu Asp Pro Gly Ala Val Gly Leu Cys Glu Phe
565 570 575
ttg aag ggg gtt caa agc gcc ttg taa 1755
Leu Lys Gly Val Gln Ser Ala Leu
580
<210>4
<211>584
<212>PRT
<213>酿酒酵母
<400>4
Met Ser Ala Lys Ser Phe Glu Val Thr Asp Pro Val Asn Ser Ser Leu
1 5 10 15
Lys Gly Phe Ala Leu Ala Asn Pro Ser Ile Thr Leu Val Pro Glu Glu
20 25 30
Lys Ile Leu Phe Arg Lys Thr Asp Ser Asp Lys Ile Ala Leu Ile Ser
35 40 45
Gly Gly Gly Ser Gly His Glu Pro Thr His Ala Gly Phe Ile Gly Lys
50 55 60
Gly Met Leu Ser Gly Ala Val Val Gly Glu Ile Phe Ala Ser Pro Ser
65 70 75 80
Thr Lys Gln Ile Leu Asn Ala Ile Arg Leu Val Asn Glu Asn Ala Ser
85 90 95
Gly Val Leu Leu Ile Val Lys Asn Tyr Thr Gly Asp Val Leu His Phe
100 105 110
Gly Leu Ser Ala Glu Arg Ala Arg Ala Leu Gly Ile Asn Cys Arg Val
115 120 125
Ala Val Ile Gly Asp Asp Val Ala Val Gly Arg Glu Lys Gly Gly Met
130 135 140
Val Gly Arg Arg Ala Leu Ala Gly Thr Val Leu Val His Lys Ile Val
145 150 155 160
Gly Ala Phe Ala Glu Glu Tyr Ser Ser Lys Tyr Gly Leu Asp Gly Thr
165 170 175
Ala Lys Val Ala Lys Ile Ile Asn Asp Asn Leu Val Thr Ile Gly Ser
180 185 190
Ser Leu Asp His Cys Lys Val Pro Gly Arg Lys Phe Glu Ser Glu Leu
195 200 205
Asn Glu Lys Gln Met Glu Leu Gly Met Gly Ile His Asn Glu Pro Gly
210 215 220
Val Lys Val Leu Asp Pro Ile Pro Ser Thr Glu Asp Leu Ile Ser Lys
225 230 235 240
Tyr Met Leu Pro Lys Leu Leu Asp Pro Asn Asp Lys Asp Arg Ala Phe
245 250 255
Val Lys Phe Asp Glu Asp Asp Glu Val Val Leu Leu Val Asn Asn Leu
260 265 270
Gly Gly Val Ser Asn Phe Val Ile Ser Ser Ile Thr Ser Lys Thr Thr
275 280 285
Asp Phe Leu Lys Glu Asn Tyr Asn Ile Thr Pro Val Gln Thr Ile Ala
290 295 300
Gly Thr Leu Met Thr Ser Phe Asn Gly Asn Gly Phe Ser Ile Thr Leu
305 310 315 320
Leu Asn Ala Thr Lys Ala Thr Lys Ala Leu Gln Ser Asp Phe Glu Glu
325 330 335
Ile Lys Ser Val Leu Asp Leu Leu Asn Ala Phe Thr Asn Ala Pro Gly
340 345 350
Trp Pro Ile Ala Asp Phe Glu Lys Thr Ser Ala Pro Ser Val Asn Asp
355 360 365
Asp Leu Leu His Asn Glu Val Thr Ala Lys Ala Val Gly Thr Tyr Asp
370 375 380
Phe Asp Lys Phe Ala Glu Trp Met Lys Ser Gly Ala Glu Gln Val Ile
385 390 395 400
Lys Ser Glu Pro His Ile Thr Glu Leu Asp Asn Gln Val Gly Asp Gly
405 410 415
Asp Cys Gly Tyr Thr Leu Val Ala Gly Val Lys Gly Ile Thr Glu Asn
420 425 430
Leu Asp Lys Leu Ser Lys Asp Ser Leu Ser Gln Ala Val Ala Gln Ile
435 440 445
Ser Asp Phe Ile Glu Gly Ser Met Gly Gly Thr Ser Gly Gly Leu Tyr
450 455 460
Ser Ile Leu Leu Ser Gly Phe Ser His Gly Leu Ile Gln Val Cys Lys
465 470 475 480
Ser Lys Asp Glu Pro Val Thr Lys Glu Ile Val Ala Lys Ser Leu Gly
485 490 495
Ile Ala Leu Asp Thr Leu Tyr Lys Tyr Thr Lys Ala Arg Lys Gly Ser
500 505 510
Ser Thr Met Ile Asp Ala Leu Glu Pro Phe Val Lys Glu Phe Thr Ala
515 520 525
Ser Lys Asp Phe Asn Lys Ala Val Lys Ala Ala Glu Glu Gly Ala Lys
530 535 540
Ser Thr Ala Thr Phe Glu Ala Lys Phe Gly Arg Ala Ser Tyr Val Gly
545 550 555 560
Asp Ser Ser Gln Val Glu Asp Pro Gly Ala Val Gly Leu Cys Glu Phe
565 570 575
Leu Lys Gly Val Gln Ser Ala Leu
580
<210>5
<211>1695
<212>DNA
<213>根瘤土壤杆菌(Agrobacterium tumefaciens)
<220>
<221>CDS
<222>(1)..(1695)
<223>
<400>5
atg aag aag ctc atc aac gat cca tcc acc gtt gtc cgg gac atg ctg 48
Met Lys Lys Leu Ile Asn Asp Pro Ser Thr Val Val Arg Asp Met Leu
1 5 10 15
gag ggc atc gtg gcg ctc agc ccc gaa acc att ctg ctt cag gat gag 96
Glu Gly Ile Val Ala Leu Ser Pro Glu Thr Ile Leu Leu Gln Asp Glu
20 25 30
aac gtg gtc ata agg tcc ggc ctg cct gaa gcg gaa aag cgc aag gtt 144
Asn Val Val Ile Arg Ser Gly Leu Pro Glu Ala Glu Lys Arg Lys Val
35 40 45
gca gtg ctt tcg ggc ggt ggc agc gga cat gag ccg gcc cac gcc ggt 192
Ala Val Leu Ser Gly Gly Gly Ser Gly His Glu Pro Ala His Ala Gly
50 55 60
tat gtc ggc acg ggc atg ttg acg gtt gcg gtg gcg ggc gat gtc ttc 240
Tyr Val Gly Thr Gly Met Leu Thr Val Ala Val Ala Gly Asp Val Phe
65 70 75 80
act tcg ccg agc acc gac gcg gtt ctc gcc ggc atc agg gcc gcg gcc 288
Thr Ser Pro Ser Thr Asp Ala Val Leu Ala Gly Ile Arg Ala Ala Ala
85 90 95
ggc cct gcc ggt gcg ctg gtc atc gtc aag aac tat acc ggc gac cgg 336
Gly Pro Ala Gly Ala Leu Val Ile Val Lys Asn Tyr Thr Gly Asp Arg
100 105 110
ctg aat ttc ggc ttg gcg gcg gag ctg gcg agg gcc gaa gga atc cct 384
Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Glu Gly Ile Pro
115 120 125
gtc gag atc gtt gtc gtc gcc gac gac gtt gcc ttg aag gat acg gtt 432
Val Glu Ile Val Val Val Ala Asp Asp Val Ala Leu Lys Asp Thr Val
130 135 140
ccc gcc gag cgc cgc cgc ggg att gcg ggg acg gtg ctc gtg cac aag 480
Pro Ala Glu Arg Arg Arg Gly Ile Ala Gly Thr Val Leu Val His Lys
145 150 155 160
ctc gcg ggg gcc gca gcg gaa aag ggc ctt cct ctt caa gag gtg gcc 528
Leu Ala Gly Ala Ala Ala Glu Lys Gly Leu Pro Leu Gln Glu Val Ala
165 170 175
cgc atc gcc cgg gac gct gcc gcc aag cta tct tcc atg ggt gtc tcg 576
Arg Ile Ala Arg Asp Ala Ala Ala Lys Leu Ser Ser Met Gly Val Ser
180 185 190
ctg gga tcc tgc acg ctg ccg gct gtc ggc aag ccg ggc ttc gtg ctt 624
Leu Gly Ser Cys Thr Leu Pro Ala Val Gly Lys Pro Gly Phe Val Leu
195 200 205
ggc gag acc gaa atc gaa gtc ggg ctc ggt atc cat ggc gag cag ggc 672
Gly Glu Thr Glu Ile Glu Val Gly Leu Gly Ile His Gly Glu Gln Gly
210 215 220
gtg cag cgg atg ccc att gcc tcg gct gac gcg ctc gtg cag ctg gtg 720
Val Gln Arg Met Pro Ile Ala Ser Ala Asp Ala Leu Val Gln Leu Val
225 230 235 240
atc gaa acg atc gaa gcc gac ggc aag ctc gcc ggc ggc aat cgc gtc 768
Ile Glu Thr Ile Glu Ala Asp Gly Lys Leu Ala Gly Gly Asn Arg Val
245 250 255
gct ctg ctg gtc aac ggc ctg ggg gca acg ccg ccg atg gaa ctc gcc 816
Ala Leu Leu Val Asn Gly Leu Gly Ala Thr Pro Pro Met Glu Leu Ala
260 265 270
atc gtc gca cgg tcg gca gtc gcg cgg ctg gag gcg aaa ggc atc gtc 864
Ile Val Ala Arg Ser Ala Val Ala Arg Leu Glu Ala Lys Gly Ile Val
275 280 285
gtg gaa cgt gcc tgg gcc ggc acc ttc ctt tca gcc ctc gat atg ccc 912
Val Glu Arg Ala Trp Ala Gly Thr Phe Leu Ser Ala Leu Asp Met Pro
290 295 300
ggg ttt tcg ttg tcg gtc atg cag gtt gac gac gca gcg ctc agc ctc 960
Gly Phe Ser Leu Ser Val Met Gln Val Asp Asp Ala Ala Leu Ser Leu
305 310 315 320
atc gac gcg cca acc gag gct ggc gca tgg ccg cgc ggc ggt gcg gtg 1008
Ile Asp Ala Pro Thr Glu Ala Gly Ala Trp Pro Arg Gly Gly Ala Val
325 330 335
aac cgc aag cgg gtt ctg cct tcg gca aac gcc gaa aag acc gtg gtt 1056
Asn Arg Lys Arg Val Leu Pro Ser Ala Asn Ala Glu Lys Thr Val Val
340 345 350
gcg aca aac aag atg acg gcg gcc ggc gag cgg ctt cgt tcg ggc gcg 1104
Ala Thr Asn Lys Met Thr Ala Ala Gly Glu Arg Leu Arg Ser Gly Ala
355 360 365
gaa cgg tcc gcg aga gcc ctg atc gct gcg gag ccc agg ctg acg caa 1152
Glu Arg Ser Ala Arg Ala Leu Ile Ala Ala Glu Pro Arg Leu Thr Gln
370 375 380
ctc gat agc gtt gca ggt gac ggc gac ctc ggc gcc agc atg gtg cgt 1200
Leu Asp Ser Val Ala Gly Asp Gly Asp Leu Gly Ala Ser Met Val Arg
385 390 395 400
ggc ggc gag gcg atc ctt gcg ctg ccg aaa gag agt ttc ggc gac gtc 1248
Gly Gly Glu Ala Ile Leu Ala Leu Pro Lys Glu Ser Phe Gly Asp Val
405 410 415
tcc gat gga ttg atg gcg atg gcc aat gcg atg cgc aag gcc atc ggc 1296
Ser Asp Gly Leu Met Ala Met Ala Asn Ala Met Arg Lys Ala Ile Gly
420 425 430
gga agc tcg ggg ccg ttc tat gcg aca ggc ctc atg cgc gct tcg cga 1344
Gly Ser Ser Gly Pro Phe Tyr Ala Thr Gly Leu Met Arg Ala Ser Arg
435 440 445
cag ctg gca ggg atc gat gag cca gcg gcc cag cag atg gcg gaa gca 1392
Gln Leu Ala Gly Ile Asp Glu Pro Ala Ala Gln Gln Met Ala Glu Ala
450 455 460
ttc gtg gcg gct gtt gcg gcg gtc tcg gaa ctt ggc ggt gcg aaa ccg 1440
Phe Val Ala Ala Val Ala Ala Val Ser Glu Leu Gly Gly Ala Lys Pro
465 470 475 480
ggc gat cgc acg atg atc gat gcg ctt tat ccg gca gcg aaa acc ttc 1488
Gly Asp Arg Thr Met Ile Asp Ala Leu Tyr Pro Ala Ala Lys Thr Phe
485 490 495
agg gac aag ctt gtg aca ggc gct tcg gca gaa gaa gcc tgg caa tcc 1536
Arg Asp Lys Leu Val Thr Gly Ala Ser Ala Glu Glu Ala Trp Gln Ser
500 505 510
gcg gtg gcg gcg ggc gag gtg ggc gca gag gcg aca gcg tcc atg aag 1584
Ala Val Ala Ala Gly Glu Val Gly Ala Glu Ala Thr Ala Ser Met Lys
515 520 525
cca cgg ctc ggg cgc gca agt tac ctc ggt gag cgg gcc gtc ggc cat 1632
Pro Arg Leu Gly Arg Ala Ser Tyr Leu Gly Glu Arg Ala Val Gly His
530 535 540
ccc gat ggg ggc gcg gtc gcc gtt ggc atc tgg ctc aaa gct atc gag 1680
Pro Asp Gly Gly Ala Val Ala Val Gly Ile Trp Leu Lys Ala Ile Glu
545 550 555 560
gct gcg atc tcg tga 1695
Ala Ala Ile Ser
<210>6
<211>564
<212>PRT
<213>根瘤土壤杆菌
<400>6
Met Lys Lys Leu Ile Asn Asp Pro Ser Thr Val Val Arg Asp Met Leu
1 5 10 15
Glu Gly Ile Val Ala Leu Ser Pro Glu Thr Ile Leu Leu Gln Asp Glu
20 25 30
Asn Val Val Ile Arg Ser Gly Leu Pro Glu Ala Glu Lys Arg Lys Val
35 40 45
Ala Val Leu Ser Gly Gly Gly Ser Gly His Glu Pro Ala His Ala Gly
50 55 60
Tyr Val Gly Thr Gly Met Leu Thr Val Ala Val Ala Gly Asp Val Phe
65 70 75 80
Thr Ser Pro Ser Thr Asp Ala Val Leu Ala Gly Ile Arg Ala Ala Ala
85 90 95
Gly Pro Ala Gly Ala Leu Val Ile Val Lys Asn Tyr Thr Gly Asp Arg
100 105 110
Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Glu Gly Ile Pro
115 120 125
Val Glu Ile Val Val Val Ala Asp Asp Val Ala Leu Lys Asp Thr Val
130 135 140
Pro Ala Glu Arg Arg Arg Gly Ile Ala Gly Thr Val Leu Val His Lys
145 150 155 160
Leu Ala Gly Ala Ala Ala Glu Lys Gly Leu Pro Leu Gln Glu Val Ala
165 170 175
Arg Ile Ala Arg Asp Ala Ala Ala Lys Leu Ser Ser Met Gly Val Ser
180 185 190
Leu Gly Ser Cys Thr Leu Pro Ala Val Gly Lys Pro Gly Phe Val Leu
195 200 205
Gly Glu Thr Glu Ile Glu Val Gly Leu Gly Ile His Gly Glu Gln Gly
210 215 220
Val Gln Arg Met Pro Ile Ala Ser Ala Asp Ala Leu Val Gln Leu Val
225 230 235 240
Ile Glu Thr Ile Glu Ala Asp Gly Lys Leu Ala Gly Gly Asn Arg Val
245 250 255
Ala Leu Leu Val Asn Gly Leu Gly Ala Thr Pro Pro Met Glu Leu Ala
260 265 270
Ile Val Ala Arg Ser Ala Val Ala Arg Leu Glu Ala Lys Gly Ile Val
275 280 285
Val Glu Arg Ala Trp Ala Gly Thr Phe Leu Ser Ala Leu Asp Met Pro
290 295 300
Gly Phe Ser Leu Ser Val Met Gln Val Asp Asp Ala Ala Leu Ser Leu
305 310 315 320
Ile Asp Ala Pro Thr Glu Ala Gly Ala Trp Pro Arg Gly Gly Ala Val
325 330 335
Asn Arg Lys Arg Val Leu Pro Ser Ala Asn Ala Glu Lys Thr Val Val
340 345 350
Ala Thr Asn Lys Met Thr Ala Ala Gly Glu Arg Leu Arg Ser Gly Ala
355 360 365
Glu Arg Ser Ala Arg Ala Leu Ile Ala Ala Glu Pro Arg Leu Thr Gln
370 375 380
Leu Asp Ser Val Ala Gly Asp Gly Asp Leu Gly Ala Ser Met Val Arg
385 390 395 400
Gly Gly Glu Ala Ile Leu Ala Leu Pro Lys Glu Ser Phe Gly Asp Val
405 410 415
Ser Asp Gly Leu Met Ala Met Ala Asn Ala Met Arg Lys Ala Ile Gly
420 425 430
Gly Ser Ser Gly Pro Phe Tyr Ala Thr Gly Leu Met Arg Ala Ser Arg
435 440 445
Gln Leu Ala Gly Ile Asp Glu Pro Ala Ala Gln Gln Met Ala Glu Ala
450 455 460
Phe Val Ala Ala Val Ala Ala Val Ser Glu Leu Gly Gly Ala Lys Pro
465 470 475 480
Gly Asp Arg Thr Met Ile Asp Ala Leu Tyr Pro Ala Ala Lys Thr Phe
485 490 495
Arg Asp Lys Leu Val Thr Gly Ala Ser Ala Glu Glu Ala Trp Gln Ser
500 505 510
Ala Val Ala Ala Gly Glu Val Gly Ala Glu Ala Thr Ala Ser Met Lys
515 520 525
Pro Arg Leu Gly Arg Ala Ser Tyr Leu Gly Glu Arg Ala Val Gly His
530 535 540
Pro Asp Gly Gly Ala Val Ala Val Gly Ile Trp Leu Lys Ala Ile Glu
545 550 555 560
Ala Ala Ile Ser
<210>7
<211>1659
<212>DNA
<213>弗氏柠檬酸杆菌(Citrobacter freundii)
<220>
<221>CDS
<222>(1)..(1659)
<223>
<400>7
atg tct caa ttc ttt ttt aac caa cgc acc cat ctt gtg agc gac gtc 48
Met Ser Gln Phe Phe Phe Asn Gln Arg Thr His Leu Val Ser Asp Val
1 5 10 15
atc gac ggg gcg att atc gcc agc cca tgg aat aac ctg gcg cgt ctg 96
Ile Asp Gly Ala Ile Ile Ala Ser Pro Trp Asn Asn Leu Ala Arg Leu
20 25 30
gaa agc gat ccg gcc att cgc atc gtg gtc cgt cgt gac ctt aat aaa l44
Glu Ser Asp Pro Ala Ile Arg Ile Val Val Arg Arg Asp Leu Asn Lys
35 40 45
aat aac gta gcg gtc att tcc ggc ggc ggt tcg gga cac gaa ccc gcg 192
Asn Asn Val Ala Val Ile Ser Gly Gly Gly Ser Gly His Glu Pro Ala
50 55 60
cac gtt ggg ttt atc ggt aaa ggc atg cta acc gct gcg gtc tgc ggc 240
His Val Gly Phe Ile Gly Lys Gly Met Leu Thr Ala Ala Val Cys Gly
65 70 75 80
gac gtt ttc gcc tcc ccg agc gtg gat gct gta ctg acc gcg att cag 288
Asp Val Phe Ala Ser Pro Ser Val Asp Ala Val Leu Thr Ala Ile Gln
85 90 95
gcg gtg acc ggt gag gct ggc tgt ttg ttg att gtg aaa aac tac acc 336
Ala Val Thr Gly Glu Ala Gly Cys Leu Leu Ile Val Lys Asn Tyr Thr
100 105 110
ggt gac cgt ctt aat ttc ggt ctc gcc gcc gag aag gcg cgt cgc ctt 384
Gly Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Lys Ala Arg Arg Leu
115 120 125
ggc tat aac gtt gaa atg ctg att gtc ggc gac gac atc tcc ctg ccg 432
Gly Tyr Asn Val Glu Met Leu Ile Val Gly Asp Asp Ile Ser Leu Pro
130 135 140
gat aac aaa cac cca cgt ggc att gcg gga act atc ctg gtg cat aaa 480
Asp Asn Lys His Pro Arg Gly Ile Ala Gly Thr Ile Leu Val His Lys
145 150 155 160
atc gca ggc tat ttt gcc gaa cgc ggc tat aac ctc gcc acc gtc ctg 528
Ile Ala Gly Tyr Phe Ala Glu Arg Gly Tyr Asn Leu Ala Thr Val Leu
165 170 175
cgt gaa gcg cag tac gca gcc agc aac acc ttt agc ctg ggc gta gcg 576
Arg Glu Ala Gln Tyr Ala Ala Ser Asn Thr Phe Ser Leu Gly Val Ala
180 185 190
ctt tcc agc tgt cat ctg ccg caa gaa acc gac gca gcc cct cgt cat 624
Leu Ser Ser Cys His Leu Pro Gln Glu Thr Asp Ala Ala Pro Arg His
195 200 205
cat ccg ggt cat gcg gag ctg ggt atg gga att cac ggc gaa cca ggc 672
His Pro Gly His Ala Glu Leu Gly Met Gly Ile His Gly Glu Pro Gly
210 215 220
gca tcg gtt atc gac acc caa aac agt gcg caa gtg gta aac ctg atg 720
Ala Ser Val Ile Asp Thr Gln Asn Ser Ala Gln Val Val Asn Leu Met
225 230 235 240
gtg gat aaa ctg ctg gcc gcc ctg cct gaa acc ggt cgt ctg gcg gtg 768
Val Asp Lys Leu Leu Ala Ala Leu Pro Glu Thr Gly Arg Leu Ala Val
245 250 255
atg att aat aat ctt ggc ggc gtt tcc gtg gcc gaa atg gcc atc atc 816
Met Ile Asn Asn Leu Gly Gly Val Ser Val Ala Glu Met Ala Ile Ile
260 265 270
acc cgc gaa ctc gcc agc agc ccg ctg cac tcg cgt atc gac tgg cta 864
Thr Arg Glu Leu Ala Ser Ser Pro Leu His Ser Arg Ile Asp Trp Leu
275 280 285
att ggc ccg gcc tcg ctg gtc acc gcg ctg gat atg aaa ggc ttc tca 912
Ile Gly Pro Ala Ser Leu Val Thr Ala Leu Asp Met Lys Gly Phe Ser
290 295 300
ctg acg gcc atc gtg ctg gaa gag agc atc gaa aaa gca ctg ctc acc 960
Leu Thr Ala Ile Val Leu Glu Glu Ser Ile Glu Lys Ala Leu Leu Thr
305 310 315 320
gaa gtg gaa acc agc aac tgg ccg acg ccg gtc cca ccg cgt gaa atc 1008
Glu Val Glu Thr Ser Asn Trp Pro Thr Pro Val Pro Pro Arg Glu Ile
325 330 335
acc tgc gta gtg tca tct cac gct agc gcc cgc gtg gaa ttc cag cct l056
Thr Cys Val Val Ser Ser His Ala Ser Ala Arg Val Glu Phe Gln Pro
340 345 350
tcg gca aac gcc ctg gtg gcc ggg att gtg gag ctg gtc acc gca acc 1104
Ser Ala Asn Ala Leu Val Ala Gly Ile Val Glu Leu Val Thr Ala Thr
355 360 365
ctt tcc gat ctg gag act cat ctg aat gcg ctg gac gcc aaa gtc ggc 1152
Leu Ser Asp Leu Glu Thr His Leu Asn Ala Leu Asp Ala Lys Val Gly
370 375 380
gat ggc gat acc ggt tcg acc ttt gcc gcc gcg gcg cgt gaa att gcc 1200
Asp Gly Asp Thr Gly Ser Thr Phe Ala Ala Ala Ala Arg Glu Ile Ala
385 390 395 400
agc ctg ctg cat cgc cag cag ctg ccg ctg aat aac ctt gcc acg ctg 1248
Ser Leu Leu His Arg Gln Gln Leu Pro Leu Asn Asn Leu Ala Thr Leu
405 410 415
ttc gcg ctg att ggc gaa cgt ctg acc gtg gtg atg ggc ggt tcc agc 1296
Phe Ala Leu Ile Gly Glu Arg Leu Thr Val Val Met Gly Gly Ser Ser
420 425 430
ggt gtg ctg atg tca atc ttc ttt acc gcc gcc ggg cag aaa ctg gaa 1344
Gly Val Leu Met Ser Ile Phe Phe Thr Ala Ala Gly Gln Lys Leu Glu
435 440 445
cag ggc gct aac gtt gtc gaa gcg cta aat acg ggg ctg gcg cag atg 1392
Gln Gly Ala Asn Val Val Glu Ala Leu Asn Thr Gly Leu Ala Gln Met
450 455 460
aag ttc tac ggc ggc gca gac gaa ggc gat cgc acg atg att gat gcg 1440
Lys Phe Tyr Gly Gly Ala Asp Glu Gly Asp Arg Thr Met Ile Asp Ala
465 470 475 480
ctg caa ccg gcc ctg acc tcg ctg ctc gca cag ccg aaa aat ctg cag 1488
Leu Gln Pro Ala Leu Thr Ser Leu Leu Ala Gln Pro Lys Asn Leu Gln
485 490 495
gcc gca ttc gac gcc gcg caa gcg gga gcc gaa cga acc tgt ttg tcg 1536
Ala Ala Phe Asp Ala Ala Gln Ala Gly Ala Glu Arg Thr Cys Leu Ser
500 505 510
agc aaa gcc aat gcg ggt cgc gca tcg tat ctg agc agc gaa agc ctg 1584
Ser Lys Ala Asn Ala Gly Arg Ala Ser Tyr Leu Ser Ser Glu Ser Leu
515 520 525
ctc gga aat atg gac ccc ggc gcg cag cgc cta gcg atg gtg ttt aaa 1632
Leu Gly Asn Met Asp Pro Gly Ala Gln Arg Leu Ala Met Val Phe Lys
530 535 540
gcg cta gcg gag agt gag ctg ggc taa 1659
Ala Leu Ala Glu Ser Glu Leu Gly
545 550
<210>8
<211>552
<212>PRT
<213>弗氏柠檬酸杆菌
<400>8
Met Ser Gln Phe Phe Phe Asn Gln Arg Thr His Leu Val Ser Asp Val
1 5 10 15
Ile Asp Gly Ala Ile Ile Ala Ser Pro Trp Asn Asn Leu Ala Arg Leu
20 25 30
Glu Ser Asp Pro Ala Ile Arg Ile Val Val Arg Arg Asp Leu Asn Lys
35 40 45
Asn Asn Val Ala Val Ile Ser Gly Gly Gly Ser Gly His Glu Pro Ala
50 55 60
His Val Gly Phe Ile Gly Lys Gly Met Leu Thr Ala Ala Val Cys Gly
65 70 75 80
Asp Val Phe Ala Ser Pro Ser Val Asp Ala Val Leu Thr Ala Ile Gln
85 90 95
Ala Val Thr Gly Glu Ala Gly Cys Leu Leu Ile Val Lys Asn Tyr Thr
100 105 110
Gly Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Lys Ala Arg Arg Leu
115 120 125
Gly Tyr Asn Val Glu Met Leu Ile Val Gly Asp Asp Ile Ser Leu Pro
130 135 140
Asp Asn Lys His Pro Arg Gly Ile Ala Gly Thr Ile Leu Val His Lys
145 150 155 160
Ile Ala Gly Tyr Phe Ala Glu Arg Gly Tyr Asn Leu Ala Thr Val Leu
165 170 175
Arg Glu Ala Gln Tyr Ala Ala Ser Asn Thr Phe Ser Leu Gly Val Ala
180 185 190
Leu Ser Ser Cys His Leu Pro Gln Glu Thr Asp Ala Ala Pro Arg His
195 200 205
His Pro Gly His Ala Glu Leu Gly Met Gly Ile His Gly Glu Pro Gly
2l0 215 220
Ala Ser Val Ile Asp Thr Gln Asn Ser Ala Gln Val Val Asn Leu Met
225 230 235 240
Val Asp Lys Leu Leu Ala Ala Leu Pro Glu Thr Gly Arg Leu Ala Val
245 250 255
Met Ile Asn Asn Leu Gly Gly Val Ser Val Ala Glu Met Ala Ile Ile
260 265 270
Thr Arg Glu Leu Ala Ser Ser Pro Leu His Ser Arg Ile Asp Trp Leu
275 280 285
Ile Gly Pro Ala Ser Leu Val Thr Ala Leu Asp Met Lys Gly Phe Ser
290 295 300
Leu Thr Ala Ile Val Leu Glu Glu Ser Ile Glu Lys Ala Leu Leu Thr
305 310 315 320
Glu Val Glu Thr Ser Asn Trp Pro Thr Pro Val Pro Pro Arg Glu Ile
325 330 335
Thr Cys Val Val Ser Ser His Ala Ser Ala Arg Val Glu Phe Gln Pro
340 345 350
Ser Ala Asn Ala Leu Val Ala Gly Ile Val Glu Leu Val Thr Ala Thr
355 360 365
Leu Ser Asp Leu Glu Thr His Leu Asn Ala Leu Asp Ala Lys Val Gly
370 375 380
Asp Gly Asp Thr Gly Ser Thr Phe Ala Ala Ala Ala Arg Glu Ile Ala
385 390 395 400
Ser Leu Leu His Arg Gln Gln Leu Pro Leu Asn Asn Leu Ala Thr Leu
405 410 415
Phe Ala Leu Ile Gly Glu Arg Leu Thr Val Val Met Gly Gly Ser Ser
420 425 430
Gly Val Leu Met Ser Ile Phe Phe Thr Ala Ala Gly Gln Lys Leu Glu
435 440 445
Gln Gly Ala Asn Val Val Glu Ala Leu Asn Thr Gly Leu Ala Gln Met
450 455 460
Lys Phe Tyr Gly Gly Ala Asp Glu Gly Asp Arg Thr Met Ile Asp Ala
465 470 475 480
Leu Gln Pro Ala Leu Thr Ser Leu Leu Ala Gln Pro Lys Asn Leu Gln
485 490 495
Ala Ala Phe Asp Ala Ala Gln Ala Gly Ala Glu Arg Thr Cys Leu Ser
500 505 510
Ser Lys Ala Asn Ala Gly Arg Ala Ser Tyr Leu Ser Ser Glu Ser Leu
515 520 525
Leu Gly Asn Met Asp Pro Gly Ala Gln Arg Leu Ala Met Val Phe Lys
530 535 540
Ala Leu Ala Glu Ser Glu Leu Gly
545 550
<210>9
<211>1740
<212>DNA
<213>人工序列
<220>
<223>attR-cat-attL-PtacM-SD-spacer
<400>9
tctagacgct caagttagta taaaaaagct gaacgagaaa cgtaaaatga tataaatatc 60
aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat 120
atgcagtcac tatgaatcaa ctacttagat ggtattagtg acctgtaaca gactgcagtg 180
gtcgaaaaaa aaagcccgca ctgtcaggtg cgggcttttt tctgtgttaa gcttcgacga 240
atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg 300
caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt 360
aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc 420
gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg 480
gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga 540
ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca 600
ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat 660
tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga 720
acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca 780
ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt 840
acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca 900
actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta 960
tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cggatccggc 1020
caagctagct tggctctagc tagagcgccc ggttgacgct gctagtgtta cctagcgatt 1080
tgtatcttac tgcatgttac ttcatgttgt caatacctgt ttttcgtgcg acttatcagg 1140
ctgtctactt atccggagat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata 1200
gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct 1260
ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag 1320
tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata 1380
accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg 1440
ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat 1500
taaagcttat cgatgataag ctgtcaaaca tgagaattcg aaatcaaata atgattttat 1560
tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt tttataatgc 1620
caacttagta taaaaaagca ggcttcaaga tctctcccca tccccctgtt cacaattaat 1680
catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca ggagactgcc 1740
<210>10
<211>80
<212>DNA
<213>人工序列
<220>
<223>PtacM
<220>
<221>启动子
<222>(1)..(80)
<223>PtacM
<400>10
ctccccatcc ccctgttcac aattaatcat cggctcgtat aatgtgtgga attgtgagcg 60
gataacaatt tcacacagga 80
<210>11
<211>2844
<212>DNA
<213>人工序列
<220>
<223>PtacMgldA::Cm
<400>11
tctagacgct caagttagta taaaaaagct gaacgagaaa cgtaaaatga tataaatatc 60
aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat 120
atgcagtcac tatgaatcaa ctacttagat ggtattagtg acctgtaaca gactgcagtg 180
gtcgaaaaaa aaagcccgca ctgtcaggtg cgggcttttt tctgtgttaa gcttcgacga 240
atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg 300
caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt 360
aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc 420
gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg 480
gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga 540
ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca 600
ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat 660
tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga 720
acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca 780
ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt 840
acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca 900
actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta 960
tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cggatccggc 1020
caagctagct tggctctagc tagagcgccc ggttgacgct gctagtgtta cctagcgatt 1080
tgtatcttac tgcatgttac ttcatgttgt caatacctgt ttttcgtgcg acttatcagg 1140
ctgtctactt atccggagat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata 1200
gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct 1260
ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag 1320
tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata 1380
accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg 1440
ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat 1500
taaagcttat cgatgataag ctgtcaaaca tgagaattcg aaatcaaata atgattttat 1560
tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt tttataatgc 1620
caacttagta taaaaaagca ggcttcaaga tctctcccca tccccctgtt cacaattaat 1680
catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca ggagactgcc 1740
atggaccgca ttattcaatc accgggtaaa tacatccagg gcgctgatgt gattaatcgt 1800
ctgggcgaat acctgaagcc gctggcagaa cgctggttag tggtgggtga caaatttgtt 1860
ttaggttttg ctcaatccac tgtcgagaaa agctttaaag atgctggact ggtagtagaa 1920
attgcgccgt ttggcggtga atgttcgcaa aatgagatcg accgtctgcg tggcatcgcg 1980
gagactgcgc agtgtggcgc aattctcggt atcggtggcg gaaaaaccct cgatactgcc 2040
aaagcactgg cacatttcat gggtgttccg gtagcgatcg caccgactat cgcctctacc 2100
gatgcaccgt gcagcgcatt gtctgttatc tacaccgatg agggtgagtt tgaccgctat 2160
ctgctgttgc caaataaccc gaatatggtc attgtcgaca ccaaaatcgt cgctggcgca 2220
cctgcacgtc tgttagcggc gggtatcggc gatgcgctgg caacctggtt tgaagcgcgt 2280
gcctgctctc gtagcggcgc gaccaccatg gcgggcggca agtgcaccca ggctgcgctg 2340
gcactggctg aactgtgcta caacaccctg ctggaagaag gcgaaaaagc gatgcttgct 2400
gccgaacagc atgtagtgac tccggcgctg gagcgcgtga ttgaagcgaa cacctatttg 2460
agcggtgttg gttttgaaag tggtggtctg gctgcggcgc acgcagtgca taacggcctg 2520
accgctatcc cggacgcgca tcactattat cacggtgaaa aagtggcatt cggtacgctg 2580
acgcagctgg ttctggaaaa tgcgccggtg gaggaaatcg aaaccgtagc tgcccttagc 2640
catgcggtag gtttgccaat aactctcgct caactggata ttaaagaaga tgtcccggcg 2700
aaaatgcgaa ttgtggcaga agcggcatgt gcagaaggtg aaaccattca caacatgcct 2760
ggcggcgcga cgccagatca ggtttacgcc gctctgctgg tagccgacca gtacggtcag 2820
cgtttcctgc aagagtggga ataa 2844
<210>12
<211>77
<212>DNA
<213>人工序列
<220>
<223>atL-ptac-g1dA
<400>12
acatcagcgc cctggatgta tttacccggt gattgaataa tgcggtccat ggcagtctcc 60
tgtgtgaaat tgttatc 77
<210>13
<211>77
<212>DNA
<213>人工序列
<220>
<223>atL-Ptac-fsaB1
<400>13
aacgccgcct ctgccgacgc tatcgccagc ctgctgcaac atgaactgga actgtaaatc 60
tagacgctca agttagt 77
<210>14
<211>50
<212>DNA
<213>人工序列
<220>
<223>pMW-dak1F
<400>14
tgattacgcc aagcttagga ggttaaatgt ccgctaaatc gtttgaagtc 50
<210>15
<211>52
<212>DNA
<213>人工序列
<220>
<223>pMW-dak1R
<400>15
atcctctaga gtcgacgcgg ccgctactta caaggcgctt tgaaccccct tc 52
<210>16
<211>846
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(846)
<223>
<400>16
atg agt caa aca tca acc ttg aaa ggc cag tgc att gct gaa ttc ctc 48
Met Ser Gln Thr Ser Thr Leu Lys Gly Gln Cys Ile Ala Glu Phe Leu
1 5 10 15
ggt acc ggg ttg ttg att ttc ttc ggt gtg ggt tgc gtt gca gca cta 96
Gly Thr Gly Leu Leu Ile Phe Phe Gly Val Gly Cys Val Ala Ala Leu
20 25 30
aaa gtc gct ggt gcg tct ttt ggt cag tgg gaa atc agt gtc att tgg 144
Lys Val Ala Gly Ala Ser Phe Gly Gln Trp Glu Ile Ser Val Ile Trp
35 40 45
gga ctg ggg gtg gca atg gcc atc tac ctg acc gca ggg gtt tcc ggc 192
Gly Leu Gly Val Ala Met Ala Ile Tyr Leu Thr Ala Gly Val Ser Gly
50 55 60
gcg cat ctt aat ccc gct gtt acc att gca ttg tgg ctg ttt gcc tgt 240
Ala His Leu Asn Pro Ala Val Thr Ile Ala Leu Trp Leu Phe Ala Cys
65 70 75 80
ttc gac aag cgc aaa gtt att cct ttt atc gtt tca caa gtt gcc ggc 288
Phe Asp Lys Arg Lys Val Ile Pro Phe Ile Val Ser Gln Val Ala Gly
85 90 95
gct ttc tgt gct gcg gct tta gtt tac ggg ctt tac tac aat tta ttt 336
Ala Phe Cys Ala Ala Ala Leu Val Tyr Gly Leu Tyr Tyr Asn Leu Phe
100 105 110
ttc gac ttc gag cag act cat cac att gtt cgc ggc agc gtt gaa agt 384
Phe Asp Phe Glu Gln Thr His His Ile Val Arg Gly Ser Val Glu Ser
115 120 125
gtt gat ctg gct ggc act ttc tct act tac cct aat cct cat atc aat 432
Val Asp Leu Ala Gly Thr Phe Ser Thr Tyr Pro Asn Pro His Ile Asn
130 135 140
ttt gtg cag gct ttc gca gtt gag atg gtg att acc gct att ctg atg 480
Phe Val Gln Ala Phe Ala Val Glu Met Val Ile Thr Ala Ile Leu Met
145 150 155 160
ggg ctg atc ctg gcg tta acg gac gat ggc aac ggt gta cca cgc ggc 528
Gly Leu Ile Leu Ala Leu Thr Asp Asp Gly Asn Gly Val Pro Arg Gly
165 170 175
cct ttg gct ccc ttg ctg att ggt cta ctg att gcg gtc att ggc gca 576
Pro Leu Ala Pro Leu Leu Ile Gly Leu Leu Ile Ala Val Ile Gly Ala
180 185 190
tct atg ggc cca ttg aca ggt ttt gcc atg aac cca gcg cgt gac ttc 624
Ser Met Gly Pro Leu Thr Gly Phe Ala Met Asn Pro Ala Arg Asp Phe
195 200 205
ggt ccg aaa gtc ttt gcc tgg ctg gcg ggc tgg ggc aat gtc gcc ttt 672
Gly Pro Lys Val Phe Ala Trp Leu Ala Gly Trp Gly Asn Val Ala Phe
210 215 220
acc ggc ggc aga gac att cct tac ttc ctg gtg ccg ctt ttc ggc cct 720
Thr Gly Gly Arg Asp Ile Pro Tyr Phe Leu Val Pro Leu Phe Gly Pro
225 230 235 240
atc gtt ggc gcg att gta ggt gca ttt gcc tac cgc aaa ctg att ggt 768
Ile Val Gly Ala Ile Val Gly Ala Phe Ala Tyr Arg Lys Leu Ile Gly
245 250 255
cgc cat ttg cct tgc gat atc tgt gtt gtg gaa gaa aag gaa acc aca 816
Arg His Leu Pro Cys Asp Ile Cys Val Val Glu Glu Lys Glu Thr Thr
260 265 270
act cct tca gaa caa aaa gct tcg ctg taa 846
Thr Pro Ser Glu Gln Lys Ala Ser Leu
275 280
<210>17
<211>281
<212>PRT
<213>大肠杆菌
<400>17
Met Ser Gln Thr Ser Thr Leu Lys Gly Gln Cys Ile Ala Glu Phe Leu
1 5 10 15
Gly Thr Gly Leu Leu Ile Phe Phe Gly Val Gly Cys Val Ala Ala Leu
20 25 30
Lys Val Ala Gly Ala Ser Phe Gly Gln Trp Glu Ile Ser Val Ile Trp
35 40 45
Gly Leu Gly Val Ala Met Ala Ile Tyr Leu Thr Ala Gly Val Ser Gly
50 55 60
Ala His Leu Asn Pro Ala Val Thr Ile Ala Leu Trp Leu Phe Ala Cys
65 70 75 80
Phe Asp Lys Arg Lys Val Ile Pro Phe Ile Val Ser Gln Val Ala Gly
85 90 95
Ala Phe Cys Ala Ala Ala Leu Val Tyr Gly Leu Tyr Tyr Asn Leu Phe
100 105 110
Phe Asp Phe Glu Gln Thr His His Ile Val Arg Gly Ser Val Glu Ser
115 120 125
Val Asp Leu Ala Gly Thr Phe Ser Thr Tyr Pro Asn Pro His Ile Asn
130 135 140
Phe Val Gln Ala Phe Ala Val Glu Met Val Ile Thr Ala Ile Leu Met
145 150 155 160
Gly Leu Ile Leu Ala Leu Thr Asp Asp Gly Asn Gly Val Pro Arg Gly
165 170 175
Pro Leu Ala Pro Leu Leu Ile Gly Leu Leu Ile Ala Val Ile Gly Ala
180 185 190
Ser Met Gly Pro Leu Thr Gly Phe Ala Met Asn Pro Ala Arg Asp Phe
195 200 205
Gly Pro Lys Val Phe Ala Trp Leu Ala Gly Trp Gly Asn Val Ala Phe
210 215 220
Thr Gly Gly Arg Asp Ile Pro Tyr Phe Leu Val Pro Leu Phe Gly Pro
225 230 235 240
Ile Val Gly Ala Ile Val Gly Ala Phe Ala Tyr Arg Lys Leu Ile Gly
245 250 255
Arg His Leu Pro Cys Asp Ile Cys Val Val Glu Glu Lys Glu Thr Thr
260 265 270
Thr Pro Ser Glu Gln Lys Ala Ser Leu
275 280
<210>18
<211>768
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(768)
<223>
<400>18
atg cga cat cct tta gtg atg ggt aac tgg aaa ctg aac ggc agc cgc 48
Met Arg His Pro Leu Val Met Gly Asn Trp Lys Leu Asn Gly Ser Arg
1 5 10 15
cac atg gtt cac gag ctg gtt tct aac ctg cgt aaa gag ctg gca ggt 96
His Met Val His Glu Leu Val Ser Asn Leu Arg Lys Glu Leu Ala Gly
20 25 30
gtt gct ggc tgt gcg gtt gca atc gca cca ccg gaa atg tat atc gat 144
Val Ala Gly Cys Ala Val Ala Ile Ala Pro Pro Glu Met Tyr Ile Asp
35 40 45
atg gcg aag cgc gaa gct gaa ggc agc cac atc atg ctg ggt gcg caa 192
Met Ala Lys Arg Glu Ala Glu Gly Ser His Ile Met Leu Gly Ala Gln
50 55 60
aac gtg gac ctg aac ctg tcc ggc gca ttc acc ggt gaa acc tct gct 240
Asn Val Asp Leu Asn Leu Ser Gly Ala Phe Thr Gly Glu Thr Ser Ala
65 70 75 80
gct atg ctg aaa gac atc ggc gca cag tac atc atc atc ggt cac tct 288
Ala Met Leu Lys Asp Ile Gly Ala Gln Tyr Ile Ile Ile Gly His Ser
85 90 95
gaa cgt cgt act tac cac aaa gaa tct gac gaa ctg atc gcg aaa aaa 336
Glu Arg Arg Thr Tyr His Lys Glu Ser Asp Glu Leu Ile Ala Lys Lys
100 105 110
ttc gcg gtg ctg aaa gag cag ggc ctg act ccg gtt ctg tgc atc ggt 384
Phe Ala Val Leu Lys Glu Gln Gly Leu Thr Pro Val Leu Cys Ile Gly
115 120 125
gaa acc gaa gct gaa aat gaa gcg ggc aaa act gaa gaa gtt tgc gca 432
Glu Thr Glu Ala Glu Asn Glu Ala Gly Lys Thr Glu Glu Val Cys Ala
130 135 140
cgt cag atc gac gcg gta ctg aaa act cag ggt gct gcg gca ttc gaa 480
Arg Gln Ile Asp Ala Val Leu Lys Thr Gln Gly Ala Ala Ala Phe Glu
145 150 155 160
ggt gcg gtt atc gct tac gaa cct gta tgg gca atc ggt act ggc aaa 528
Gly Ala Val Ile Ala Tyr Glu Pro ValTrp Ala Ile Gly Thr Gly Lys
165 170 175
tct gca act ccg gct cag gca cag gct gtt cac aaa ttc atc cgt gac 576
Ser Ala Thr Pro Ala Gln Ala Gln Ala Val His Lys Phe Ile Arg Asp
180 185 190
cac atc gct aaa gtt gac gct aac atc gct gaa caa gtg atc att cag 624
His Ile Ala Lys Val Asp Ala Asn Ile Ala Glu Gln Val Ile Ile Gln
195 200 205
tac ggc ggc tct gta aac gcg tct aac gct gca gaa ctg ttt gct cag 672
Tyr Gly Gly Ser Val Asn Ala Ser Asn Ala Ala Glu Leu Phe Ala Gln
210 215 220
ccg gat atc gac ggc gcg ctg gtt ggt ggt gct tct ctg aaa gct gac 720
Pro Asp Ile Asp Gly Ala Leu Val Gly Gly Ala Ser Leu Lys Ala Asp
225 230 235 240
gcc ttc gca gta atc gtt aaa gct gca gaa gcg gct aaa cag gct taa 768
Ala Phe Ala Val Ile Val Lys Ala Ala Glu Ala Ala Lys Gln Ala
245 250 255
<210>19
<211>255
<212>PRT
<213>大肠杆菌
<400>19
Met Arg His Pro Leu Val Met Gly Asn Trp Lys Leu Asn Gly Ser Arg
1 5 10 15
His Met Val His Glu Leu Val Ser Asn Leu Arg Lys Glu Leu Ala Gly
20 25 30
Val Ala Gly Cys Ala Val Ala Ile Ala Pro Pro Glu Met Tyr Ile Asp
35 40 45
Met Ala Lys Arg Glu Ala Glu Gly Ser His Ile Met Leu Gly Ala Gln
50 55 60
Asn Val Asp Leu Asn Leu Ser Gly Ala Phe Thr Gly Glu Thr Ser Ala
65 70 75 80
Ala Met Leu Lys Asp Ile Gly Ala Gln Tyr Ile Ile Ile Gly His Ser
85 90 95
Glu Arg Arg Thr Tyr His Lys Glu Ser Asp Glu Leu Ile Ala Lys Lys
100 105 110
Phe Ala Val Leu Lys Glu Gln Gly Leu Thr Pro Val Leu Cys Ile Gly
115 120 125
Glu Thr Glu Ala Glu Asn Glu Ala Gly Lys Thr Glu Glu Val Cys Ala
130 135 140
Arg Gln Ile Asp Ala Val Leu Lys Thr Gln Gly Ala Ala Ala Phe Glu
145 150 155 160
Gly Ala Val Ile Ala Tyr Glu Pro Val Trp Ala Ile Gly Thr Gly Lys
165 170 175
Ser Ala Thr Pro Ala Gln Ala Gln Ala Val His Lys Phe Ile Arg Asp
180 185 190
His Ile Ala Lys Val Asp Ala Asn Ile Ala Glu Gln Val Ile Ile Gln
195 200 205
Tyr Gly Gly Ser Val Asn Ala Ser Asn Ala Ala Glu Leu Phe Ala Gln
210 215 220
Pro Asp Ile Asp Gly Ala Leu Val Gly Gly Ala Ser Leu Lys Ala Asp
225 230 235 240
Ala Phe Ala Val Ile Val Lys Ala Ala Glu Ala Ala Lys Gln Ala
245 250 255
<210>20
<211>1080
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1080)
<223>
<400>20
atg tct aag att ttt gat ttc gta aaa cct ggc gta atc act ggt gat 48
Met Ser Lys Ile Phe Asp Phe Val Lys Pro Gly Val Ile Thr Gly Asp
1 5 10 15
gac gta cag aaa gtt ttc cag gta gca aaa gaa aac aac ttc gca ctg 96
Asp Val Gln Lys Val Phe Gln Val Ala Lys Glu Asn Asn Phe Ala Leu
20 25 30
cca gca gta aac tgc gtc ggt act gac tcc atc aac gcc gta ctg gaa 144
Pro Ala Val Asn Cys Val Gly Thr Asp Ser Ile Asn Ala Val Leu Glu
35 40 45
acc gct gct aaa gtt aaa gcg ccg gtt atc gtt cag ttc tcc aac ggt 192
Thr Ala Ala Lys Val Lys Ala Pro Val Ile Val Gln Phe Ser Asn Gly
50 55 60
ggt gct tcc ttt atc gct ggt aaa ggc gtg aaa tct gac gtt ccg cag 240
Gly Ala Ser Phe Ile Ala Gly Lys Gly Val Lys Ser Asp Val Pro Gln
65 70 75 80
ggt gct gct atc ctg ggc gcg atc tct ggt gcg cat cac gtt cac cag 288
Gly Ala Ala Ile Leu Gly Ala Ile Ser Gly Ala His His Val His Gln
85 90 95
atg gct gaa cat tat ggt gtt ccg gtt atc ctg cac act gac cac tgc 336
Met Ala Glu His Tyr Gly Val Pro Val Ile Leu His Thr Asp His Cys
100 105 110
gcg aag aaa ctg ctg ccg tgg atc gac ggt ctg ttg gac gcg ggt gaa 384
Ala Lys Lys Leu Leu Pro Trp Ile Asp Gly Leu Leu Asp Ala Gly Glu
115 120 125
aaa cac ttc gca gct acc ggt aag ccg ctg ttc tct tct cac atg atc 432
Lys His Phe Ala Ala Thr Gly Lys Pro Leu Phe Ser Ser His Met Ile
130 135 140
gac ctg tct gaa gaa tct ctg caa gag aac atc gaa atc tgc tct aaa 480
Asp Leu Ser Glu Glu Ser Leu Gln Glu AsnIle GluIle Cys Ser Lys
145 150 155 160
tac ctg gag cgc atg tcc aaa atc ggc atg act ctg gaa atc gaa ctg 528
Tyr Leu Glu Arg Met Ser Lys Ile Gly Met Thr Leu Glu Ile Glu Leu
165 170 175
ggt tgc acc ggt ggt gaa gaa gac ggc gtg gac aac agc cac atg gac 576
Gly Cys Thr Gly Gly Glu Glu Asp Gly Val Asp Asn Ser His Met Asp
180 185 190
gct tct gca ctg tac acc cag ccg gaa gac gtt gat tac gca tac acc 624
Ala Ser Ala Leu Tyr Thr Gln Pro Glu Asp Val Asp Tyr Ala Tyr Thr
195 200 205
gaa ctg agc aaa atc agc ccg cgt ttc acc atc gca gcg tcc ttc ggt 672
Glu Leu Ser Lys Ile Ser Pro Arg Phe Thr Ile Ala Ala Ser Phe Gly
210 215 220
aac gta cac ggt gtt tac aag ccg ggt aac gtg gtt ctg act ccg acc 720
Asn Val His Gly Val Tyr Lys Pro Gly Asn Val Val Leu Thr Pro Thr
225 230 235 240
atc ctg cgt gat tct cag gaa tat gtt tcc aag aaa cac aac ctg ccg 768
Ile Leu Arg Asp Ser Gln Glu Tyr Val Ser Lys Lys His Asn Leu Pro
245 250 255
cac aac agc ctg aac ttc gta ttc cac ggt ggt tcc ggt tct act gct 816
His Asn Ser Leu Asn Phe Val Phe His Gly Gly Ser Gly Ser Thr Ala
260 265 270
cag gaa atc aaa gac tcc gta agc tac ggc gta gta aaa atg aac atc 864
Gln Glu Ile Lys Asp Ser Val Ser Tyr Gly Val Val Lys Met AsnIle
275 280 285
gat acc gat acc caa tgg gca acc tgg gaa ggc gtt ctg aac tac tac 912
Asp Thr Asp Thr Gln Trp Ala Thr Trp Glu Gly Val Leu Asn Tyr Tyr
290 295 300
aaa gcg aac gaa gct tat ctg cag ggt cag ctg ggt aac ccg aaa ggc 960
Lys Ala Asn Glu Ala Tyr Leu Gln Gly Gln Leu Gly Asn Pro Lys Gly
305 310 315 320
gaa gat cag ccg aac aag aaa tac tac gat ccg cgc gta tgg ctg cgt 1008
Glu Asp Gln Pro Asn Lys Lys Tyr Tyr Asp Pro Arg Val Trp Leu Arg
325 330 335
gcc ggt cag act tcg atg atc gct cgt ctg gag aaa gca ttc cag gaa 1056
Ala Gly Gln Thr Ser Met Ile Ala Arg Leu Glu Lys Ala Phe Gln Glu
340 345 350
ctg aac gcg atc gac gtt ctg taa 1080
Leu Asn Ala Ile Asp Val Leu
355
<210>21
<211>359
<212>PRT
<213>大肠杆菌
<400>21
Met Ser Lys Ile Phe Asp Phe Val Lys Pro Gly Val Ile Thr Gly Asp
1 5 10 15
Asp Val Gln Lys Val Phe Gln Val Ala Lys Glu Ash Asn Phe Ala Leu
20 25 30
Pro Ala Val Asn Cys Val Gly Thr Asp Ser Ile Asn Ala Val Leu Glu
35 40 45
Thr Ala Ala Lys Val Lys Ala Pro Val Ile Val Gln Phe Ser Asn Gly
50 55 60
Gly Ala Ser Phe Ile Ala Gly Lys Gly Val Lys Ser Asp Val Pro Gln
65 70 75 80
Gly Ala Ala Ile Leu Gly Ala Ile Ser Gly Ala His His Val His Gln
85 90 95
Met Ala Glu His Tyr Gly Val Pro Val Ile Leu His Thr Asp His Cys
100 105 110
Ala Lys Lys Leu Leu Pro Trp Ile Asp Gly Leu Leu Asp Ala Gly Glu
115 120 125
Lys His Phe Ala Ala Thr Gly Lys Pro Leu Phe Ser Ser His Met Ile
130 135 140
Asp Leu Ser Glu Glu Ser Leu Gln Glu Asn Ile Glu Ile Cys Ser Lys
145 150 155 160
Tyr Leu Glu Arg Met Ser Lys Ile Gly Met Thr Leu Glu Ile Glu Leu
165 170 175
Gly Cys Thr Gly Gly Glu Glu Asp Gly Val Asp Asn Ser His Met Asp
180 185 190
Ala Ser Ala Leu Tyr Thr Gln Pro Glu Asp Val Asp Tyr Ala Tyr Thr
195 200 205
Glu Leu Ser Lys Ile Ser Pro Arg Phe Thr Ile Ala Ala Ser Phe Gly
210 215 220
Asn Val His Gly Val Tyr Lys Pro Gly Asn Val Val Leu Thr Pro Thr
225 230 235 240
Ile Leu Arg Asp Ser Gln Glu Tyr Val Ser Lys Lys His Asn Leu Pro
245 250 255
His Asn Ser Leu Ash Phe Val Phe His Gly Gly Ser Gly Ser Thr Ala
260 265 270
Gln Glu Ile Lys Asp Ser Val Ser Tyr Gly Val Val Lys Met Asn Ile
275 280 285
Asp Thr Asp Thr Gln Trp Ala Thr Trp Glu Gly Val Leu Asn Tyr Tyr
290 295 300
Lys Ala Asn Glu Ala Tyr Leu Gln Gly Gln Leu Gly Asn Pro Lys Gly
305 310 315 320
Glu Asp Gln Pro Ash Lys Lys Tyr Tyr Asp Pro Arg Val Trp Leu Arg
325 330 335
Ala Gly Gln Thr Ser Met Ile Ala Arg Leu Glu Lys Ala Phe Gln Glu
340 345 350
Leu Asn Ala Ile Asp Val Leu
355
<210>22
<211>1011
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1011)
<223>
<400>22
atg aga cga gaa ctt gcc atc gaa ttt tcc cgc gtc acc gaa tca gcg 48
Met Arg Arg Glu Leu Ala Ile Glu Phe Ser Arg Val Thr Glu Ser Ala
1 5 10 15
gcg ctg gct ggc tac aaa tgg tta gga cgc ggc gat aaa aac acc gcg 96
Ala Leu Ala Gly Tyr Lys Trp Leu Gly Arg Gly Asp Lys Asn Thr Ala
20 25 30
gac ggc gcg gcg gta aac gcc atg cgt att atg ctc aac cag gtc aac 144
Asp Gly Ala Ala Val Asn Ala Met Arg Ile Met Leu Asn Gln Val Asn
35 40 45
att gac ggc acc atc gtc att ggt gaa ggt gaa atc gac gaa gca ccg 192
Ile Asp Gly Thr Ile Val Ile Gly Glu Gly Glu Ile Asp Glu Ala Pro
50 55 60
atg ctc tac att ggt gaa aaa gtc ggt act ggt cgc ggc gac gcg gta 240
Met Leu Tyr Ile Gly Glu Lys Val Gly Thr Gly Arg Gly Asp Ala Val
65 70 75 80
gat att gct gtt gat ccg att gaa ggc acg cgc atg acg gcg atg ggc 288
Asp Ile Ala Val Asp Pro Ile Glu Gly Thr Arg Met Thr Ala Met Gly
85 90 95
cag gct aac gcg ctg gcg gtg ctg gca gta ggc gat aaa ggc tgc ttc 336
Gln Ala Asn Ala Leu Ala Val Leu Ala Val Gly Asp Lys Gly Cys Phe
100 105 110
ctc aat gcg ccg gat atg tat atg gag aag ctg att gtc ggg ccg gga 384
Leu Asn Ala Pro Asp Met Tyr Met Glu Lys Leu Ile Val Gly Pro Gly
115 120 125
gcc aaa ggc acc att gat ctg aac ctg ccg ctg gcg gat aac ctg cgc 432
Ala Lys Gly Thr Ile Asp Leu Asn Leu Pro Leu Ala Asp Asn Leu Arg
130 135 140
aat gta gcg gcg gcg ctc ggc aaa ccg ttg agc gaa ctg acg gta acg 480
Asn Val Ala Ala Ala Leu Gly Lys Pro Leu Ser Glu Leu Thr Val Thr
145 150 155 160
att ctg gct aaa cca cgc cac gat gcc gtt atc gct gaa atg cag caa 528
Ile Leu Ala Lys Pro Arg His Asp Ala Val Ile Ala Glu Met Gln Gln
165 170 175
ctc ggc gta cgc gta ttt gct att ccg gac ggc gac gtt gcg gcc tca 576
Leu Gly Val Arg Val Phe Ala Ile Pro Asp Gly Asp Val Ala Ala Ser
180 185 190
att ctc acc tgt atg cca gac agc gaa gtt gac gtg ctg tac ggt att 624
Ile Leu Thr Cys Met Pro Asp Ser Glu Val Asp Val Leu Tyr Gly Ile
195 200 205
ggt ggc gcg ccg gaa ggc gta gtt tct gcg gcg gtg atc cgc gca tta 672
Gly Gly Ala Pro Glu Gly Val Val Ser Ala Ala Val Ile Arg Ala Leu
210 215 220
gat ggc gac atg aac ggt cgt ctg ctg gcg cgt cat gac gtc aaa ggc 720
Asp Gly Asp Met Asn Gly Arg Leu Leu Ala Arg His Asp Val Lys Gly
225 230 235 240
gac aac gaa gag aat cgt cgc att ggc gag cag gag ctg gca cgc tgc 768
Asp Asn Glu Glu Asn Arg Arg Ile Gly Glu Gln Glu Leu Ala Arg Cys
245 250 255
aaa gcg atg ggc atc gaa gcc ggt aaa gta ttg cgc ctg ggc gat atg 816
Lys Ala Met Gly Ile Glu Ala Gly Lys Val Leu Arg Leu Gly Asp Met
260 265 270
gcg cgc agc gat aac gtc atc ttc tct gcc acc ggt att acc aaa ggc 864
Ala Arg Ser Asp Asn Val Ile Phe Ser Ala Thr Gly Ile Thr Lys Gly
275 280 285
gat ctg ctg gaa ggc att agc cgc aaa ggc aat atc gcg act acc gaa 912
Asp Leu Leu Glu Gly Ile Ser Arg Lys Gly Asn Ile Ala Thr Thr Glu
290 295 300
acg ctg ctg atc cgc ggc aag tca cgc acc att cgc cgc att cag tcc 960
Thr Leu Leu Ile Arg Gly Lys Ser Arg Thr Ile Arg Arg Ile Gln Ser
305 310 315 320
atc cac tat ctg gat cgc aaa gac ccg gaa atg cag gtg cac atc ctc 1008
Ile His Tyr Leu Asp Arg Lys Asp Pro Glu Met Gln Val His Ile Leu
325 330 335
tga 1011
<210>23
<211>336
<212>PRT
<213>大肠杆菌
<400>23
Met Arg Arg Glu Leu Ala Ile Glu Phe Ser Arg Val Thr Glu Ser Ala
1 5 10 15
Ala Leu Ala Gly Tyr Lys Trp Leu Gly Arg Gly Asp Lys Asn Thr Ala
20 25 30
Asp Gly Ala Ala Val Asn Ala Met Arg Ile Met Leu Asn Gln Val Asn
35 40 45
Ile Asp Gly Thr Ile Val Ile Gly Glu Gly Glu Ile Asp Glu Ala Pro
50 55 60
Met Leu Tyr Ile Gly Glu Lys Val Gly Thr Gly Arg Gly Asp Ala Val
65 70 75 80
Asp Ile Ala Val Asp Pro Ile Glu Gly Thr Arg Met Thr Ala Met Gly
85 90 95
Gln Ala Asn Ala Leu Ala Val Leu Ala Val Gly Asp Lys Gly Cys Phe
100 105 110
Leu Asn Ala Pro Asp Met Tyr Met Glu Lys Leu Ile Val Gly Pro Gly
115 120 125
Ala Lys Gly Thr Ile Asp Leu Asn Leu Pro Leu Ala Asp Asn Leu Arg
130 135 140
Asn Val Ala Ala Ala Leu Gly Lys Pro Leu Ser Glu Leu Thr Val Thr
145 150 155 160
Ile Leu Ala Lys Pro Arg His Asp Ala Val Ile Ala Glu Met Gln Gln
165 170 175
Leu Gly Val Arg Val Phe Ala Ile Pro Asp Gly Asp Val Ala Ala Ser
180 185 190
Ile Leu Thr Cys Met Pro Asp Ser Glu Val Asp Val Leu Tyr Gly Ile
195 200 205
Gly Gly Ala Pro Glu Gly Val Val Ser Ala Ala Val Ile Arg Ala Leu
210 215 220
Asp Gly Asp Met Asn Gly Arg Leu Leu Ala Arg His Asp Val Lys Gly
225 230 235 240
Asp Asn Glu Glu Asn Arg Arg Ile Gly Glu Gln Glu Leu Ala Arg Cys
245 250 255
Lys Ala Met Gly Ile Glu Ala Gly Lys Val Leu Arg Leu Gly Asp Met
260 265 270
Ala Arg Ser Asp Asn Val Ile Phe Ser Ala Thr Gly Ile Thr Lys Gly
275 280 285
Asp Leu Leu Glu Gly Ile Ser Arg Lys Gly Asn Ile Ala Thr Thr Glu
290 295 300
Thr Leu Leu Ile Arg Gly Lys Ser Arg Thr Ile Arg Arg Ile Gln Ser
305 310 315 320
Ile His Tyr Leu Asp Arg Lys Asp Pro Glu Met Gln Val His Ile Leu
325 330 335
<210>24
<211>1509
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1509)
<223>
<400>24
atg act gaa aaa aaa tat atc gtt gcg ctc gac cag ggc acc acc agc 48
Met Thr Glu Lys Lys Tyr Ile Val Ala Leu Asp Gln Gly Thr Thr Ser
1 5 10 15
tcc cgc gcg gtc gta atg gat cac gat gcc aat atc att agc gtg tcg 96
Ser Arg Ala Val Val Met Asp His Asp Ala Asn Ile Ile Ser Val Ser
20 25 30
cag cgc gaa ttt gag caa atc tac cca aaa cca ggt tgg gta gaa cac 144
Gln Arg Glu Phe Glu Gln Ile Tyr Pro Lys Pro Gly Trp Val Glu His
35 40 45
gac cca atg gaa atc tgg gcc acc caa agc tcc acg ctg gta gaa gtg 192
Asp Pro Met Glu Ile Trp Ala Thr Gln Ser Ser Thr Leu Val Glu Val
50 55 60
ctg gcg aaa gcc gat atc agt tcc gat caa att gca gct atc ggt att 240
Leu Ala Lys Ala Asp Ile Ser Ser Asp Gln Ile Ala Ala Ile Gly Ile
65 70 75 80
acg aac cag cgt gaa acc act att gtc tgg gaa aaa gaa acc ggc aag 288
Thr Asn Gln Arg Glu Thr Thr Ile Val Trp Glu Lys Glu Thr Gly Lys
85 90 95
cct atc tat aac gcc att gtc tgg cag tgc cgt cgt acc gca gaa atc 336
Pro Ile Tyr Asn Ala Ile Val Trp Gln Cys Arg Arg Thr Ala Glu Ile
100 105 110
tgc gag cat tta aaa cgt gac ggt tta gaa gat tat atc cgc agc aat 384
Cys Glu His Leu Lys Arg Asp Gly Leu Glu Asp Tyr Ile Arg Ser Asn
115 120 125
acc ggt ctg gtg att gac ccg tac ttt tct ggc acc aaa gtg aag tgg 432
Thr Gly Leu Val Ile Asp Pro Tyr Phe Ser Gly Thr Lys Val Lys Trp
130 135 140
atc ctc gac cat gtg gaa ggc tct cgc gag cgt gca cgt cgt ggt gaa 480
Ile Leu Asp His Val Glu Gly Ser Arg Glu Arg Ala Arg Arg Gly Glu
145 150 155 160
ttg ctg ttt ggt acg gtt gat acg tgg ctt atc tgg aaa atg act cag 528
Leu Leu Phe Gly Thr Val Asp Thr Trp Leu Ile Trp Lys Met Thr Gln
165 170 175
ggc cgt gtc cat gtg acc gat tac acc aac gcc tct cgt acc atg ttg 576
Gly Arg Val His Val Thr Asp Tyr Thr Asn Ala Ser Arg Thr Met Leu
180 185 190
ttc aac atc cat acc ctg gac tgg gac gac aaa atg ctg gaa gtg ctg 624
Phe Asn Ile His Thr Leu Asp Trp Asp Asp Lys Met Leu Glu Val Leu
195 200 205
gat att ccg cgc gag atg ctg cca gaa gtg cgt cgt tct tcc gaa gta 672
Asp Ile Pro Arg Glu Met Leu Pro Glu Val Arg Arg Ser Ser Glu Val
210 215 220
tac ggt cag act aac att ggc ggc aaa ggc ggc acg cgt att cca atc 720
Tyr Gly Gln Thr Asn Ile Gly Gly Lys Gly Gly Thr Arg Ile Pro Ile
225 230 235 240
tcc ggg atc gcc ggt gac cag cag gcc gcg ctg ttt ggt cag ttg tgc 768
Ser Gly Ile Ala Gly Asp Gln Gln Ala Ala Leu Phe Gly Gln Leu Cys
245 250 255
gtg aaa gaa ggg atg gcg aag aac acc tat ggc act ggc tgc ttt atg 816
Val Lys Glu Gly Met Ala Lys Asn Thr Tyr Gly Thr Gly Cys Phe Met
260 265 270
ctg atg aac act ggc gag aaa gcg gtg aaa tca gaa aac ggc ctg ctg 864
Leu Met Asn Thr Gly Glu Lys Ala Val Lys Ser Glu Asn Gly Leu Leu
275 280 285
acc acc atc gcc tgc ggc ccg act ggc gaa gtg aac tat gcg ttg gaa 912
Thr Thr Ile Ala Cys Gly Pro Thr Gly Glu Val Asn Tyr Ala Leu Glu
290 295 300
ggt gcg gtg ttt atg gca ggc gca tcc att cag tgg ctg cgc gat gaa 960
Gly Ala Val Phe Met Ala Gly Ala Ser Ile Gln Trp Leu Arg Asp Glu
305 310 315 320
atg aag ttg att aac gac gcc tac gat tcc gaa tat ttc gcc acc aaa 1008
Met Lys Leu Ile Asn Asp Ala Tyr Asp Ser Glu Tyr Phe Ala Thr Lys
325 330 335
gtg caa aac acc aat ggt gtg tat gtg gtt ccg gca ttt acc ggg ctg 1056
Val Gln Asn Thr Asn Gly Val Tyr Val Val Pro Ala Phe Thr Gly Leu
340 345 350
ggt gcg ccg tac tgg gac ccg tat gcg cgc ggg gcg att ttc ggt ctg 1104
Gly Ala Pro Tyr Trp Asp Pro Tyr Ala Arg Gly Ala Ile Phe Gly Leu
355 360 365
act cgt ggg gtg aac gct aac cac att ata cgc gcg acg ctg gag tct 1152
Thr Arg Gly Val Asn Ala Asn His Ile Ile Arg Ala Thr Leu Glu Ser
370 375 380
att gct tat cag acg cgt gac gtg ctg gaa gcg atg cag gcc gac tct 1200
Ile Ala Tyr Gln Thr Arg Asp Val Leu Glu Ala Met Gln Ala Asp Ser
385 390 395 400
ggt atc cgt ctg cac gcc ctg cgc gtg gat ggt ggc gca gta gca aac 1248
Gly Ile Arg Leu His Ala Leu Arg Val Asp Gly Gly Ala Val Ala Asn
405 410 415
aat ttc ctg atg cag ttc cag tcc gat att ctc ggc acc cgc gtt gag 1296
Asn Phe Leu Met Gln Phe Gln Ser Asp Ile Leu Gly Thr Arg Val Glu
420 425 430
cgc ccg gaa gtg cgc gaa gtc acc gca ttg ggt gcg gcc tat ctc gca 1344
Arg Pro Glu Val Arg Glu Val Thr Ala Leu Gly Ala Ala Tyr Leu Ala
435 440 445
ggc ctg gcg gtt ggc ttc tgg cag aac ctc gac gag ctg caa gag aaa 1392
Gly Leu Ala Val Gly Phe Trp Gln Asn Leu Asp Glu Leu Gln Glu Lys
450 455 460
gcg gtg att gag cgc gag ttc cgt cca ggc atc gaa acc act gag cgt 1440
Ala Val Ile Glu Arg Glu Phe Arg Pro Gly Ile Glu Thr Thr Glu Arg
465 470 475 480
aat tac cgt tac gca ggc tgg aaa aaa gcg gtt aaa cgc gcg atg gcg 1488
Asn Tyr Arg Tyr Ala Gly Trp Lys Lys Ala Val Lys Arg Ala Met Ala
485 490 495
tgg gaa gaa cac gac gaa taa 1509
Trp Glu Glu His Asp Glu
500
<210>25
<211>502
<212>PRT
<213>大肠杆菌
<400>25
Met Thr Glu Lys Lys Tyr Ile Val Ala Leu Asp Gln Gly Thr Thr Ser
1 5 10 15
Ser Arg Ala Val Val Met Asp His Asp Ala Asn Ile Ile Ser Val Ser
20 25 30
Gln Arg Glu Phe Glu Gln Ile Tyr Pro Lys Pro Gly Trp Val Glu His
35 40 45
Asp Pro Met Glu Ile Trp Ala Thr Gln Ser Ser Thr Leu Val Glu Val
50 55 60
Leu Ala Lys Ala Asp Ile Ser Ser Asp Gln Ile Ala Ala Ile Gly Ile
65 70 75 80
Thr Asn Gln Arg Glu Thr Thr Ile Val Trp Glu Lys Glu Thr Gly Lys
85 90 95
Pro Ile Tyr Asn Ala Ile Val Trp Gln Cys Arg Arg Thr Ala Glu Ile
100 105 110
Cys Glu His Leu Lys Arg Asp Gly Leu Glu Asp Tyr Ile Arg Ser Asn
115 120 125
Thr Gly Leu Val Ile Asp Pro Tyr Phe Ser Gly Thr Lys Val Lys Trp
130 135 140
Ile Leu Asp His Val Glu Gly Ser Arg Glu Arg Ala Arg Arg Gly Glu
145 150 155 160
Leu Leu Phe Gly Thr Val Asp Thr Trp Leu Ile Trp Lys Met Thr Gln
165 170 175
Gly Arg Val His Val Thr Asp Tyr Thr Asn Ala Ser Arg Thr Met Leu
180 185 190
Phe Asn Ile His Thr Leu Asp Trp Asp Asp Lys Met Leu Glu Val Leu
195 200 205
Asp Ile Pro Arg Glu Met Leu Pro Glu Val Arg Arg Ser Ser Glu Val
210 215 220
Tyr Gly Gln Thr Asn Ile Gly Gly Lys Gly Gly Thr Arg Ile Pro Ile
225 230 235 240
Ser Gly Ile Ala Gly Asp Gln Gln Ala Ala Leu Phe Gly Gln Leu Cys
245 250 255
Val Lys Glu Gly Met Ala Lys Asn Thr Tyr Gly Thr Gly Cys Phe Met
260 265 270
Leu Met Asn Thr Gly Glu Lys Ala Val Lys Ser Glu Asn Gly Leu Leu
275 280 285
Thr Thr Ile Ala Cys Gly Pro Thr Gly Glu Val Asn Tyr Ala Leu Glu
290 295 300
Gly Ala Val Phe Met Ala Gly Ala Ser Ile Gln Trp Leu Arg Asp Glu
305 310 315 320
Met Lys Leu Ile Asn Asp Ala Tyr Asp Ser Glu Tyr Phe Ala Thr Lys
325 330 335
Val Gln Asn Thr Asn Gly Val Tyr Val Val Pro Ala Phe Thr Gly Leu
340 345 350
Gly Ala Pro Tyr Trp Asp Pro Tyr Ala Arg Gly Ala Ile Phe Gly Leu
355 360 365
Thr Arg Gly Val Asn Ala Asn His Ile Ile Arg Ala Thr Leu Glu Ser
370 375 380
Ile Ala Tyr Gln Thr Arg Asp Val Leu Glu Ala Met Gln Ala Asp Ser
385 390 395 400
Gly Ile Arg Leu His Ala Leu Arg Val Asp Gly Gly Ala Val Ala Asn
405 410 415
Asn Phe Leu Met Gln Phe Gln Ser Asp Ile Leu Gly Thr Arg Val Glu
420 425 430
Arg Pro Glu Val Arg Glu Val Thr Ala Leu Gly Ala Ala Tyr Leu Ala
435 440 445
Gly Leu Ala Val Gly Phe Trp Gln Asn Leu Asp Glu Leu Gln Glu Lys
450 455 460
Ala Val Ile Glu Arg Glu Phe Arg Pro Gly Ile Glu Thr Thr Glu Arg
465 470 475 480
Ash Tyr Arg Tyr Ala Gly Trp Lys Lys Ala Val Lys Arg Ala Met Ala
485 490 495
Trp Glu Glu His Asp Glu
500
<210>26
<211>1629
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1629)
<223>
<400>26
atg aaa act cgc gac tcg caa tca agt gac gtg att atc att ggc ggc 48
Met Lys Thr Arg Asp Ser Gln Ser Ser Asp Val Ile Ile Ile Gly Gly
1 5 10 15
ggc gca acg gga gcc ggg att gcc cgc gac tgt gcc ctg cgc ggg ctg 96
Gly Ala Thr Gly Ala Gly Ile Ala Arg Asp Cys Ala Leu Arg Gly Leu
20 25 30
cgc gtg att ttg gtt gag cgc cac gac arc gca acc ggt gcc acc ggg 144
Arg Val Ile Leu Val Glu Arg His Asp Ile Ala Thr Gly Ala Thr Gly
35 40 45
cgt aac cac ggc ctg ctg cac agc ggt gcg cgc tat gcg gta acc gat 192
Arg Asn His Gly Leu Leu His Ser Gly Ala Arg Tyr Ala Val Thr Asp
50 55 60
gcg gaa tcg gcc cgc gaa tgc att agt gaa aac cag atc ctg aaa cgc 240
Ala Glu Ser Ala Arg Glu Cys Ile Ser Glu Asn Gln Ile Leu Lys Arg
65 70 75 80
att gca cgt cac tgc gtt gaa cca acc aac ggc ctg ttt atc acc ctg 288
Ile Ala Arg His Cys Val Glu Pro Thr Asn Gly Leu Phe Ile Thr Leu
85 90 95
ccg gaa gat gac ctc tcc ttc cag gcc act ttt att cgc gcc tgc gaa 336
Pro Glu Asp Asp Leu Ser Phe Gln Ala Thr Phe Ile Arg Ala Cys Glu
100 105 110
gaa gca ggg atc agc gca gaa gct ata gac ccg cag caa gcg cgc att 384
Glu Ala Gly Ile Ser Ala Glu Ala Ile Asp Pro Gln Gln Ala Arg Ile
115 120 125
atc gaa cct gcc gtt aac ccg gca ctg att ggc gcg gtg aaa gtt ccg 432
Ile Glu Pro Ala Val Asn Pro Ala Leu Ile Gly Ala Val Lys Val Pro
130 135 140
gat ggc acc gtt gat cca ttt cgt ctg acc gca gca aac atg ctg gat 480
Asp Gly Thr Val Asp Pro Phe Arg Leu Thr Ala Ala Asn Met Leu Asp
145 150 155 160
gcc aaa gaa cac ggt gcc gtt atc ctt acc gct cat gaa gtc acg ggg 528
Ala Lys Glu His Gly Ala Val Ile Leu Thr Ala His Glu Val Thr Gly
165 170 175
ctg att cgt gaa ggc gcg acg gtg tgc ggt gtt cgt gta cgt aac cat 576
Leu Ile Arg Glu Gly Ala Thr Val Cys Gly Val Arg Val Arg Asn His
180 185 190
ctc acc ggc gaa act cag gcc ctt cat gca cct gtc gtg gtt aat gcc 624
Leu Thr Gly Glu Thr Gln Ala Leu His Ala Pro Val Val Val Asn Ala
195 200 205
gct ggg atc tgg ggg caa cac att gcc gaa tat gcc gat ctg cgc att 672
Ala Gly Ile Trp Gly Gln His Ile Ala Glu Tyr Ala Asp Leu Arg Ile
210 215 220
cgc atg ttc ccg gcg aaa gga tcg ctg ctg atc atg gat cac cgc att 720
Arg Met Phe Pro Ala Lys Gly Ser Leu Leu Ile Met Asp His Arg Ile
225 230 235 240
aac cag cat gtg atc aac cgc tgc cgt aaa cct tcc gac gcc gat att 768
Asn Gln His Val Ile Asn Arg Cys Arg Lys Pro Ser Asp Ala Asp Ile
245 250 255
ctg gtg cct ggc gat acc att tcg ctg att ggt acc acc tct tta cgt 816
Leu Val Pro Gly Asp Thr Ile Ser Leu Ile Gly Thr Thr Ser Leu Arg
260 265 270
att gat tac aac gag att gac gat aat cga gtg acg gca gaa gag gtt 864
Ile Asp Tyr Asn Glu Ile Asp Asp Asn Arg Val Thr Ala Glu Glu Val
275 280 285
gat att ctg ctg cgt gaa ggg gaa aaa ctg gcc ccc gtg atg gcg aaa 912
Asp Ile Leu Leu Arg Glu Gly Glu Lys Leu Ala Pro Val Met Ala Lys
290 295 300
acg cgc att ttg cgg gcc tat tct ggc gtg cgc ccg ctg gtt gcc agc 960
Thr Arg Ile Leu Arg Ala Tyr Ser Gly Val Arg Pro Leu Val Ala Ser
305 310 315 320
gat gac gac ccg agc gga cgt aac gtc agc cgt ggc atc gtg ctg ctc 1008
Asp Asp Asp Pro Ser Gly Arg Asn Val Ser Arg Gly Ile Val Leu Leu
325 330 335
gac cat gct gaa cgc gat ggt ctg gac gga ttt atc acc atc acc ggt 1056
Asp His Ala Glu Arg Asp Gly Leu Asp Gly Phe Ile Thr Ile Thr Gly
340 345 350
ggc aaa ctg atg acc tat cgg ctg atg gct gaa tgg gct acc gac gcg 1104
Gly Lys Leu Met Thr Tyr Arg Leu Met Ala Glu Trp Ala Thr Asp Ala
355 360 365
gta tgc cgc aaa ctg ggc aac acg cgc ccc tgt acg act gcc gat ctg 1152
Val Cys Arg Lys Leu Gly Asn Thr Arg Pro Cys Thr Thr Ala Asp Leu
370 375 380
gca ctg cct ggt tca caa gaa ccc gct gaa gtt acc ttg cgt aaa gtc 1200
Ala Leu Pro Gly Ser Gln Glu Pro Ala Glu Val Thr Leu Arg Lys Val
385 390 395 400
atc tcc ctg cct gcc ccg ctg cgc ggt tct gcg gtt tat cgt cat ggc 1248
Ile Ser Leu Pro Ala Pro Leu Arg Gly Ser Ala Val Tyr Arg His Gly
405 410 415
gat cgc acg cct gcc tgg ctg agc gaa ggc cgt ctg cac cgt agc ctg 1296
Asp Arg Thr Pro Ala Trp Leu Ser Glu Gly Arg Leu His Arg Ser Leu
420 425 430
gta tgt gag tgc gaa gcg gta act gcg ggt gaa gtg cag tac gcg gta 1344
Val Cys Glu Cys Glu Ala Val Thr Ala Gly Glu Val Gln Tyr Ala Val
435 440 445
gaa aat tta aac gtt aat agc ctg ctg gat tta cgc cgt cgt acc cgt 1392
Glu Asn Leu Asn Val Asn Ser Leu Leu Asp Leu Arg Arg Arg Thr Arg
450 455 460
gtg ggg atg ggc acc tgc cag ggc gaa ctc tgc gcc tgc cgc gct gcc 1440
Val Gly Met Gly Thr Cys Gln Gly Glu Leu Cys Ala Cys Arg Ala Ala
465 470 475 480
gga ctg ctg caa cgt ttt aac gtc acg acg tcc gcg caa tct atc gag 1488
Gly Leu Leu Gln Arg Phe Asn Val Thr Thr Ser Ala Gln Ser Ile Glu
485 490 495
caa ctt tcc acc ttc ctt aac gaa cgc tgg aaa ggc gtg caa ccc atc 1536
Gln Leu Ser Thr Phe Leu Asn Glu Arg Trp Lys Gly Val Gln Pro Ile
500 505 510
gcc tgg gga gat gca ctg cgc gaa agc gaa ttt acc cgc tgg gtt tat 1584
Ala Trp Gly Asp Ala Leu Arg Glu Ser Glu Phe Thr Arg Trp Val Tyr
515 520 525
cag gga ttg tgt ggt ctg gag aag gag cag aaa gat gcg ctt tga 1629
Gln Gly Leu Cys Gly Leu Glu Lys Glu Gln Lys Asp Ala Leu
530 535 540
<210>27
<211>542
<212>PRT
<213>大肠杆菌
<400>27
Met Lys Thr Arg Asp Ser Gln Ser Ser Asp Val Ile Ile Ile Gly Gly
1 5 10 15
Gly Ala Thr Gly Ala Gly Ile Ala Arg Asp Cys Ala Leu Arg Gly Leu
20 25 30
Arg Val Ile Leu Val Glu Arg His Asp Ile Ala Thr Gly Ala Thr Gly
35 40 45
Arg Asn His Gly Leu Leu His Ser Gly Ala Arg Tyr Ala Val Thr Asp
50 55 60
Ala Glu Ser Ala Arg Glu Cys Ile Ser Glu Asn Gln Ile Leu Lys Arg
65 70 75 80
Ile Ala Arg His Cys Val Glu Pro Thr Asn Gly Leu Phe Ile Thr Leu
85 90 95
Pro Glu Asp Asp Leu Ser Phe Gln Ala Thr Phe Ile Arg Ala Cys Glu
100 105 110
Glu Ala Gly Ile Ser Ala Glu Ala Ile Asp Pro Gln Gln Ala Arg Ile
115 120 125
Ile Glu Pro Ala Val Asn Pro Ala Leu Ile Gly Ala Val Lys Val Pro
130 135 140
Asp Gly Thr Val Asp Pro Phe Arg Leu Thr Ala Ala Asn Met Leu Asp
145 150 155 160
Ala Lys Glu His Gly Ala Val Ile Leu Thr Ala His Glu Val Thr Gly
165 170 175
Leu Ile Arg Glu Gly Ala Thr Val Cys Gly Val Arg Val Arg Asn His
180 185 190
Leu Thr Gly Glu Thr Gln Ala Leu His Ala Pro Val Val Val Asn Ala
195 200 205
Ala Gly Ile Trp Gly Gln His Ile Ala Glu Tyr Ala Asp Leu Arg Ile
210 215 220
Arg Met Phe Pro Ala Lys Gly Ser Leu Leu Ile Met Asp His Arg Ile
225 230 235 240
Asn Gln His Val Ile Asn Arg Cys Arg Lys Pro Ser Asp Ala Asp Ile
245 250 255
Leu Val Pro Gly Asp Thr Ile Ser Leu Ile Gly Thr Thr Ser Leu Arg
260 265 270
Ile Asp Tyr Asn Glu Ile Asp Asp Asn Arg Val Thr Ala Glu Glu Val
275 280 285
Asp Ile Leu Leu Arg Glu Gly Glu Lys Leu Ala Pro Val Met Ala Lys
290 295 300
Thr Arg Ile Leu Arg Ala Tyr Ser Gly Val Arg Pro Leu Val Ala Ser
305 310 315 320
Asp Asp Asp Pro Ser Gly Arg Asn Val Ser Arg Gly Ile Val Leu Leu
325 330 335
Asp His Ala Glu Arg Asp Gly Leu Asp Gly Phe Ile Thr Ile Thr Gly
340 345 350
Gly Lys Leu Met Thr Tyr Arg Leu Met Ala Glu Trp Ala Thr Asp Ala
355 360 365
Val Cys Arg Lys Leu Gly Asn Thr Arg Pro Cys Thr Thr Ala Asp Leu
370 375 380
Ala Leu Pro Gly Ser Gln Glu Pro Ala Glu Val Thr Leu Arg Lys Val
385 390 395 400
Ile Ser Leu Pro Ala Pro Leu Arg Gly Ser Ala Val Tyr Arg His Gly
405 410 415
Asp Arg Thr Pro Ala Trp Leu Ser Glu Gly Arg Leu His Arg Ser Leu
420 425 430
Val Cys Glu Cys Glu Ala Val Thr Ala Gly Glu Val Gln Tyr Ala Val
435 440 445
Glu Asn Leu Asn Val Asn Ser Leu Leu Asp Leu Arg Arg Arg Thr Arg
450 455 460
Val Gly Met Gly Thr Cys Gln Gly Glu Leu Cys Ala Cys Arg Ala Ala
465 470 475 480
Gly Leu Leu Gln Arg Phe Asn Val Thr Thr Ser Ala Gln Ser Ile Glu
485 490 495
Gln Leu Ser Thr Phe Leu Asn Glu Arg Trp Lys Gly Val Gln Pro Ile
500 505 510
Ala Trp Gly Asp Ala Leu Arg Glu Ser Glu Phe Thr Arg Trp Val Tyr
515 520 525
Gln Gly Leu Cys Gly Leu Glu Lys Glu Gln Lys Asp Ala Leu
530 535 540
<210>28
<211>1260
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1260)
<223>
<400>28
atg cgc ttt gat act gtc att atg ggc ggc ggc ctc gcc gga tta ctc 48
Met Arg Phe Asp Thr Val Ile Met Gly Gly Gly Leu Ala Gly Leu Leu
1 5 10 15
tgt ggc ctg caa ctg caa aaa cac ggc ctg cgc tgt gcc att gtc act 96
Cys Gly Leu Gln Leu Gln Lys His Gly Leu Arg Cys Ala Ile Val Thr
20 25 30
cgt ggt caa agc gca ctg cat ttc tca tcc gga tcg ctg gat ttg ctg 144
Arg Gly Gln Ser Ala Leu His Phe Ser Ser Gly Ser Leu Asp Leu Leu
35 40 45
agc cat ctg cca gat ggt caa ccg gtg aca gac att cac agt gga ctg 192
Ser His Leu Pro Asp Gly Gln Pro Val Thr Asp Ile His Ser Gly Leu
50 55 60
gaa tct ttg cgt cag cag gca cca gcc cat cct tac tcc ctt ctc gag 240
Glu Ser Leu Arg Gln Gln Ala Pro Ala His Pro Tyr Ser Leu Leu Glu
65 70 75 80
cca caa cgc gtg ctc gat ctc gct tgc cag gcg cag gca tta atc gct 288
Pro Gln Arg Val Leu Asp Leu Ala Cys Gln Ala Gln Ala Leu Ile Ala
85 90 95
gaa agc ggt gcg caa ttg cag ggc agc gta gaa ctt gct cac cag cgg 336
Glu Ser Gly Ala Gln Leu Gln Gly Ser Val Glu Leu Ala His Gln Arg
100 105 110
gtt acg ccg ctc ggc act ctg cgc tct acc tgg cta agt tcg cca gaa 384
Val Thr Pro Leu Gly Thr Leu Arg Ser Thr Trp Leu Ser Ser Pro Glu
115 120 125
gtc ccc gtc tgg ccg ctg ccc gcg aag aaa ata tgt gta gtg gga att 432
Val Pro Val Trp Pro Leu Pro Ala Lys Lys Ile Cys Val Val Gly Ile
130 135 140
agc ggc ctg atg gat ttt cag gcg cac ctt gcg gca gct tcg ttg cgt 480
Ser Gly Leu Met Asp Phe Gln Ala His Leu Ala Ala Ala Ser Leu Arg
145 150 155 160
gaa ctc ggc ctt gcc gtt gaa acc gca gaa ata gag ctg ccg gaa ctg 528
Glu Leu Gly Leu Ala Val Glu Thr Ala Glu Ile Glu Leu Pro Glu Leu
165 170 175
gat gtg ctg cgc aat aac gcc acc gaa ttt cgc gcg gtg aat atc gcc 576
Asp Val Leu Arg Asn Asn Ala Thr Glu Phe Arg Ala Val Asn Ile Ala
180 185 190
cgt ttc ctt gat aat gaa gaa aac tgg ccg ctg tta ctt gat gcg ctt 624
Arg Phe Leu Asp Asn Glu Glu Asn Trp Pro Leu Leu Leu Asp Ala Leu
195 200 205
att cct gtc gcc aat acc tgc gaa atg atc ctg atg ccc gcc tgc ttc 672
Ile Pro Val Ala Asn Thr Cys Glu Met Ile Leu Met Pro Ala Cys Phe
210 215 220
ggt ctg gcc gat gac aaa ctg tgg cgt tgg ttg aat gaa aaa cta cct 720
Gly Leu Ala Asp Asp Lys Leu Trp Arg Trp Leu Asn Glu Lys Leu Pro
225 230 235 240
tgt tca ctg atg ctt ttg cca acg ctg ccg cct tcc gtg ctg ggc att 768
Cys Ser Leu Met Leu Leu Pro Thr Leu Pro Pro Ser Val Leu Gly Ile
245 250 255
cgt ctg caa aac cag tta cag cgc cag ttt gtg cgc cag ggt ggc gtg 816
Arg Leu Gln Asn Gln Leu Gln Arg Gln Phe Val Arg Gln Gly Gly Val
260 265 270
tgg atg ccg ggc gat gaa gtg aaa aaa gtg acc tgt aaa aat ggc gta 864
Trp Met Pro Gly Asp Glu Val Lys Lys Val Thr Cys Lys Asn Gly Val
275 280 285
gtg aac gaa atc tgg acc cgc aat cac gcc gat att ccg cta cgt cca 912
Val Asn Glu Ile Trp Thr Arg Asn His Ala Asp Ile Pro Leu Arg Pro
290 295 300
cgt ttc gcg gtt ctc gcc agc ggc agt ttc ttt agt ggc gga ctg gta 960
Arg Phe Ala Val Leu Ala Ser Gly Ser Phe Phe Ser Gly Gly Leu Val
305 310 315 320
gcg gaa cgt aac ggc att cga gag ccg att ctc ggc ctt gat gtg cta 1008
Ala Glu Arg Asn Gly Ile Arg Glu Pro Ile Leu Gly Leu Asp Val Leu
325 330 335
caa acc gcc acg cgg ggt gaa tgg tat aag gga gat ttt ttt gcg ccg 1056
Gln Thr Ala Thr Arg Gly Glu Trp Tyr Lys Gly Asp Phe Phe Ala Pro
340 345 350
caa ccg tgg cag cag ttc ggt gta acc act gat gag acg cta cgc ccg 1104
Gln Pro Trp Gln Gln Phe Gly Val Thr Thr Asp Glu Thr Leu Arg Pro
355 360 365
tca cag gca ggg caa acc att gaa aac ctg ttt gcc atc ggt tcg gtg 1152
Ser Gln Ala Gly Gln Thr Ile Glu Asn Leu Phe Ala Ile Gly Ser Val
370 375 380
ctg ggc gga ttt gat ccc atc gcc cag gga tgc ggc ggc ggt gtt tgt 1200
Leu Gly Gly Phe Asp Pro Ile Ala Gln Gly Cys Gly Gly Gly Val Cys
385 390 395 400
gcc gtc agt gct tta cat gcc gct caa cag att gcc caa cgc gca gga 1248
Ala Val Ser Ala Leu His Ala Ala Gln Gln Ile Ala Gln Arg Ala Gly
405 410 415
ggc caa caa tga 1260
Gly Gln Gln
<210>29
<211>419
<212>PRT
<213>大肠杆菌
<400>29
Met Arg Phe Asp Thr Val Ile Met Gly Gly Gly Leu Ala Gly Leu Leu
1 5 10 15
Cys Gly Leu Gln Leu Gln Lys His Gly Leu Arg Cys Ala Ile Val Thr
20 25 30
Arg Gly Gln Ser Ala Leu His Phe Ser Ser Gly Ser Leu Asp Leu Leu
35 40 45
Ser His Leu Pro Asp Gly Gln Pro Val Thr Asp Ile His Ser Gly Leu
50 55 60
Glu Ser Leu Arg Gln Gln Ala Pro Ala His Pro Tyr Ser Leu Leu Glu
65 70 75 80
Pro Gln Arg Val Leu Asp Leu Ala Cys Gln Ala Gln Ala Leu Ile Ala
85 90 95
Glu Ser Gly Ala Gln Leu Gln Gly Ser Val Glu Leu Ala His Gln Arg
100 105 110
Val Thr Pro Leu Gly Thr Leu Arg Ser Thr Trp Leu Ser Ser Pro Glu
115 120 125
Val Pro Val Trp Pro Leu Pro Ala Lys Lys Ile Cys Val Val Gly Ile
130 135 140
Ser Gly Leu Met Asp Phe Gln Ala His Leu Ala Ala Ala Ser Leu Arg
145 150 155 160
Glu Leu Gly Leu Ala Val Glu Thr Ala Glu Ile Glu Leu Pro Glu Leu
165 170 175
Asp Val Leu Arg Asn Asn Ala Thr Glu Phe Arg Ala Val Asn Ile Ala
180 185 190
Arg Phe Leu Asp Asn Glu Glu Asn Trp Pro Leu Leu Leu Asp Ala Leu
195 200 205
Ile Pro Val Ala Asn Thr Cys Glu Met Ile Leu Met Pro Ala Cys Phe
210 215 220
Gly Leu Ala Asp Asp Lys Leu Trp Arg Trp Leu Asn Glu Lys Leu Pro
225 230 235 240
Cys Ser Leu Met Leu Leu Pro Thr Leu Pro Pro Ser Val Leu Gly Ile
245 250 255
Arg Leu Gln Asn Gln Leu Gln Arg Gln Phe Val Arg Gln Gly Gly Val
260 265 270
Trp Met Pro Gly Asp Glu Val Lys Lys Val Thr Cys Lys Asn Gly Val
275 280 285
Val Asn Glu Ile Trp Thr Arg Asn His Ala Asp Ile Pro Leu Arg Pro
290 295 300
Arg Phe Ala Val Leu Ala Ser Gly Ser Phe Phe Ser Gly Gly Leu Val
305 310 315 320
Ala Glu Arg Asn Gly Ile Arg Glu Pro Ile Leu Gly Leu Asp Val Leu
325 330 335
Gln Thr Ala Thr Arg Gly Glu Trp Tyr Lys Gly Asp Phe Phe Ala Pro
340 345 350
Gln Pro Trp Gln Gln Phe Gly Val Thr Thr Asp Glu Thr Leu Arg Pro
355 360 365
Ser Gln Ala Gly Gln Thr Ile Glu Asn Leu Phe Ala Ile Gly Ser Val
370 375 380
Leu Gly Gly Phe Asp Pro Ile Ala Gln Gly Cys Gly Gly Gly Val Cys
385 390 395 400
Ala Val Ser Ala Leu His Ala Ala Gln Gln Ile Ala Gln Arg Ala Gly
405 410 415
Gly Gln Gln
<210>30
<211>1191
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1191)
<223>
<400>30
atg aat gac acc agc ttc gaa aac tgc att aag tgc acc gtc tgc acc 48
Met Asn Asp Thr Ser Phe Glu Asn Cys Ile Lys Cys Thr Val Cys Thr
1 5 10 15
acc gcc tgc ccg gtg agc cgg gtg aat ccc ggt tat cca ggg cca aaa 96
Thr Ala Cys Pro Val Ser Arg Val Asn Pro Gly Tyr Pro Gly Pro Lys
20 25 30
caa gcc ggg ccg gat ggc gag cgt ctg cgt ttg aaa gat ggc gca ctg 144
Gln Ala Gly Pro Asp Gly Glu Arg Leu Arg Leu Lys Asp Gly Ala Leu
35 40 45
tat gac gag gcg ctg aaa tat tgc atc aac tgc aaa cgt tgt gaa gtc 192
Tyr Asp Glu Ala Leu Lys Tyr Cys Ile Asn Cys Lys Arg Cys Glu Val
50 55 60
gcc tgc ccg tcc gat gtg aag att ggc gat att atc cag cgc gcg cgg 240
Ala Cys Pro Ser Asp Val Lys Ile Gly Asp Ile Ile Gln Arg Ala Arg
65 70 75 80
gcg aaa tat gac acc acg cgc ccg tcg ctg cgt aat ttt gtg ttg agt 288
Ala Lys Tyr Asp Thr Thr Arg Pro Ser Leu Arg Asn Phe Val Leu Ser
85 90 95
cat acc gac ctg atg ggt agc gtt tcc acg ccg ttc gca cca atc gtc 336
His Thr Asp Leu Met Gly Ser Val Ser Thr Pro Phe Ala Pro Ile Val
100 105 110
aac acc gct acc tcg ctg aaa ccg gtg cgg cag ctg ctt gat gcg gcg 384
Asn Thr Ala Thr Ser Leu Lys Pro Val Arg Gln Leu Leu Asp Ala Ala
115 120 125
tta aaa atc gat cat cgc cgc acg cta ccg aaa tac tcc ttc ggc acg 432
Leu Lys Ile Asp His Arg Arg Thr Leu Pro Lys Tyr Ser Phe Gly Thr
130 135 140
ttc cgt cgc tgg tat cgc agc gtg geg gct cag caa gca caa tat aaa 480
Phe Arg Arg Trp Tyr Arg Ser Val Ala Ala Gln Gln Ala Gln Tyr Lys
145 150 155 160
gac cag gtc gct ttc ttt cac ggc tgc ttc gtt aac tac aac cat ccg 528
Asp Gln Val Ala Phe Phe His Gly Cys Phe Val Asn Tyr Asn His Pro
165 170 175
cag tta ggt aaa gat tta att aaa gtg ctc aac gca atg ggt acc ggt 576
Gln Leu Gly Lys Asp Leu Ile Lys Val Leu Asn Ala Met Gly Thr Gly
180 185 190
gta caa ctg ctc agc aaa gaa aaa tgc tgc ggc gta ccg cta atc gcc 624
Val Gln Leu Leu Ser Lys Glu Lys Cys Cys Gly Val Pro Leu Ile Ala
195 200 205
aac ggc ttt acc gat aaa gca cgc aaa cag gca att acg aat gta gag 672
Asn Gly Phe Thr Asp Lys Ala Arg Lys Gln Ala Ile Thr Asn Val Glu
2l0 215 220
tcg atc cgc gaa gct gtg gga gta aaa ggc att ccg gtg att gcc acc 720
Ser Ile Arg Glu Ala Val Gly Val Lys Gly Ile Pro Val Ile Ala Thr
225 230 235 240
tcc tca acc tgt aca ttt gcc ctg cgc gac gaa tac ccg gaa gtg ctg 768
Ser Ser Thr Cys Thr Phe Ala Leu Arg Asp Glu Tyr Pro Glu Val Leu
245 250 255
aat gtc gac aac aaa ggc ttg cgc gat cat atc gaa ctg gca acc cgc 816
Asn Val Asp Asn Lys Gly Leu Arg Asp His Ile Glu Leu Ala Thr Arg
260 265 270
tgg ctg tgg cgc aag ctg gac gaa ggc aaa acg tta ccg ctg aaa ccg 864
Trp Leu Trp Arg Lys Leu Asp Glu Gly Lys Thr Leu Pro Leu Lys Pro
275 280 285
ctg ccg ctg aaa gtg gtt tat cac act ccg tgc cat atg gaa aaa atg 912
Leu Pro Leu Lys Val Val Tyr His Thr Pro Cys His Met Glu Lys Met
290 295 300
ggc tgg acg ctc tac acc ctg gag ctg ttg cgt aac atc ccg ggg ctt 960
Gly Trp Thr Leu Tyr Thr Leu Glu Leu Leu Arg Asn Ile Pro Gly Leu
305 310 315 320
gag tta acg gtg ctg gat tcc cag tgc tgc ggt att gcg ggt act tac 1008
Glu Leu Thr Val Leu Asp Ser Gln Cys Cys Gly Ile Ala Gly Thr Tyr
325 330 335
ggt ttc aaa aaa gag aac tac ccc acc tca caa gcc atc ggc gca cca 1056
Gly Phe Lys Lys Glu Asn Tyr Pro Thr Ser Gln Ala Ile Gly Ala Pro
340 345 350
ctg ttc cgc cag ata gaa gaa agc ggc gca gat ctg gtg gtc acc gac 1104
Leu Phe Arg Gln Ile Glu Glu Ser Gly Ala Asp Leu Val Val Thr Asp
355 360 365
tgc gaa acc tgt aaa tgg cag att gag atg tcc aca agt ctt cgc tgc 1152
Cys Glu Thr Cys Lys Trp Gln Ile Glu Met Ser Thr Ser Leu Arg Cys
370 375 380
gaa cat ccg att acg cta ctg gcc cag gcg ctg gct taa 1191
Glu His Pro Ile Thr Leu Leu Ala Gln Ala Leu Ala
385 390 395
<210>31
<211>396
<212>PRT
<213>大肠杆菌
<400>31
Met Asn Asp Thr Ser Phe Glu Asn Cys Ile Lys Cys Thr Val Cys Thr
1 5 10 15
Thr Ala Cys Pro Val Ser Arg Val Asn Pro Gly Tyr Pro Gly Pro Lys
20 25 30
Gln Ala Gly Pro Asp Gly Glu Arg Leu Arg Leu Lys Asp Gly Ala Leu
35 40 45
Tyr Asp Glu Ala Leu Lys Tyr Cys Ile Asn Cys Lys Arg Cys Glu Val
50 55 60
Ala Cys Pro Ser Asp Val Lys Ile Gly Asp Ile Ile Gln Arg Ala Arg
65 70 75 80
Ala Lys Tyr Asp Thr Thr Arg Pro Ser Leu Arg Asn Phe Val Leu Ser
85 90 95
His Thr Asp Leu Met Gly Ser Val Ser Thr Pro Phe Ala Pro Ile Val
100 105 110
Asn Thr Ala Thr Ser Leu Lys Pro Val Arg Gln Leu Leu Asp Ala Ala
115 120 125
Leu Lys Ile Asp His Arg Arg Thr Leu Pro Lys Tyr Ser Phe Gly Thr
130 135 140
Phe Arg Arg Trp Tyr Arg Ser Val Ala Ala Gln Gln Ala Gln Tyr Lys
145 150 155 160
Asp Gln Val Ala Phe Phe His Gly Cys Phe Val Asn Tyr Asn His Pro
165 170 175
Gln Leu Gly Lys Asp Leu Ile Lys Val Leu Asn Ala Met Gly Thr Gly
180 185 190
Val Gln Leu Leu Ser Lys Glu Lys Cys Cys Gly Val Pro Leu Ile Ala
195 200 205
Asn Gly Phe Thr Asp Lys Ala Arg Lys Gln Ala Ile Thr Asn Val Glu
210 215 220
Ser Ile Arg Glu Ala Val Gly Val Lys Gly Ile Pro Val Ile Ala Thr
225 230 235 240
Ser Ser Thr Cys Thr Phe Ala Leu Arg Asp Glu Tyr Pro Glu Val Leu
245 250 255
Asn Val Asp Asn Lys Gly Leu Arg Asp His Ile Glu Leu Ala Thr Arg
260 265 270
Trp Leu Trp Arg Lys Leu Asp Glu Gly Lys Thr Leu Pro Leu Lys Pro
275 280 285
Leu Pro Leu Lys Val Val Tyr His Thr Pro Cys His Met Glu Lys Met
290 295 300
Gly Trp Thr Leu Tyr Thr Leu Glu Leu Leu Arg Asn Ile Pro Gly Leu
305 310 315 320
Glu Leu Thr Val Leu Asp Ser Gln Cys Cys Gly Ile Ala Gly Thr Tyr
325 330 335
Gly Phe Lys Lys Glu Asn Tyr Pro Thr Ser Gln Ala Ile Gly Ala Pro
340 345 350
Leu Phe Arg Gln Ile Glu Glu Ser Gly Ala Asp Leu Val Val Thr Asp
355 360 365
Cys Glu Thr Cys Lys Trp Gln Ile Glu Met Ser Thr Ser Leu Arg Cys
370 375 380
Glu His Pro Ile Thr Leu Leu Ala Gln Ala Leu Ala
385 390 395
<210>32
<211>1506
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1506)
<223>
<400>32
atg gaa acc aaa gat ctg att gtg ata ggg ggc ggc atc aat ggt gct 48
Met Glu Thr Lys Asp Leu Ile Val Ile Gly Gly Gly Ile Asn Gly Ala
1 5 10 15
ggt atc gcg gca gac gcc gct gga cgc ggt tta tcc gtg ctg atg ctg 96
Gly Ile Ala Ala Asp Ala Ala Gly Arg Gly Leu Ser Val Leu Met Leu
20 25 30
gag gcg cag gat ctc gct tgc gcg acc tct tcc gcc agt tca aaa ctc 144
Glu Ala Gln Asp Leu Ala Cys Ala Thr Ser Ser Ala Ser Ser Lys Leu
35 40 45
att cac ggt ggc ctg cgc tac ctt gag cac tat gaa ttc cgc ctg gtc 192
Ile His Gly Gly Leu Arg Tyr Leu Glu His Tyr Glu Phe Arg Leu Val
50 55 60
agc gag gcg ctg gct gaa cgt gaa gtg ctg ctg aaa atg gcc ccg cat 240
Ser Glu Ala Leu Ala Glu Arg Glu Val Leu Leu Lys Met Ala Pro His
65 70 75 80
atc gcc ttc ccg atg cgt ttt cgc ctg cca cat cgt ccg cat ctg cgc 288
Ile Ala Phe Pro Met Arg Phe Arg Leu Pro His Arg Pro His Leu Arg
85 90 95
ccg gcg tgg atg att cgc att ggt ctg ttt atg tac gat cat ctg ggt 336
Pro Ala Trp Met Ile Arg Ile Gly Leu Phe Met Tyr Asp His Leu Gly
100 105 110
aaa cgc acc agc ttg ccg gga tca act ggt ttg cgt ttt ggc gca aat 384
Lys Arg Thr Ser Leu Pro Gly Ser Thr Gly Leu Arg Phe Gly Ala Asn
115 120 125
tca gtg tta aaa ccg gaa att aag cgc gga ttc gaa tat tct gac tgt 432
Ser Val Leu Lys Pro Glu Ile Lys Arg Gly Phe Glu Tyr Ser Asp Cys
130 135 140
tgg gta gac gac gcc cgt ctg gta ctc gcc aac gcc cag atg gtg gtg 480
Trp Val Asp Asp Ala Arg Leu Val Leu Ala Asn Ala Gln Met Val Val
145 150 155 160
cgt aaa ggc ggc gaa gtg ctt act cgg act cgc gcc acc tct gct cgc 528
Arg Lys Gly Gly Glu Val Leu Thr Arg Thr Arg Ala Thr Ser Ala Arg
165 170 175
cgc gaa aac ggc ctg tgg att gtg gaa gcg gaa gat atc gat acc ggc 576
Arg Glu Asn Gly Leu Trp Ile Val Glu Ala Glu Asp Ile Asp Thr Gly
180 185 190
aaa aaa tat agc tgg caa gcg cgc ggc ttg gtt aac gcc acc ggc ccg 624
Lys Lys Tyr Ser Trp Gln Ala Arg Gly Leu Val Asn Ala Thr Gly Pro
195 200 205
tgg gtg aaa cag ttc ttc gac gac ggg atg cat ctg cct tcg cct tat 672
Trp Val Lys Gln Phe Phe Asp Asp Gly Met His Leu Pro Ser Pro Tyr
210 215 220
ggc att cgc ctg atc aaa ggc agc cat att gtg gtg ccg cgc gtg cat 720
Gly Ile Arg Leu Ile Lys Gly Ser His Ile Val Val Pro Arg Val His
225 230 235 240
acc cag aag caa gcc tac att ctg caa aac gaa gat aaa cgt att gtg 768
Thr Gln Lys Gln Ala Tyr Ile Leu Gln Asn Glu Asp Lys Arg Ile Val
245 250 255
ttc gtg atc ccg tgg atg gac gag ttt tcc atc atc ggc act acc gat 816
Phe Val Ile Pro Trp Met Asp Glu Phe Ser Ile Ile Gly Thr Thr Asp
260 265 270
gtc gag tac aaa ggc gat ccg aaa gcg gtg aag att gaa gag agt gaa 864
Val Glu Tyr Lys Gly Asp Pro Lys Ala Val Lys Ile Glu Glu Ser Glu
275 280 285
atc aat tac ctg ctg aat gtg tat aac acg cac ttt aaa aag cag tta 912
Ile Asn Tyr Leu Leu Asn Val Tyr Asn Thr His Phe Lys Lys Gln Leu
290 295 300
agc cgt gac gat atc gtc tgg acc tac tcc ggt gtg cgt ccg ctg tgt 960
Ser Arg Asp Asp Ile Val Trp Thr Tyr Ser Gly Val Arg Pro Leu Cys
305 310 315 320
gat gat gag tcc gac tcg ccg cag gct att acc cgt gat tac acc ctt 1008
Asp Asp Glu Ser Asp Ser Pro Gln Ala Ile Thr Arg Asp Tyr Thr Leu
325 330 335
gat att cat gat gaa aat ggc aaa gca ccg ctg ctg tcg gta ttc ggc 1056
Asp Ile His Asp Glu Asn Gly Lys Ala Pro Leu Leu Ser Val Phe Gly
340 345 350
ggt aag ctg acc acc tac cga aaa ctg gcg gaa cat gcg ctg gaa aaa 1104
Gly Lys Leu Thr Thr Tyr Arg Lys Leu Ala Glu His Ala Leu Glu Lys
355 360 365
cta acg ccg tat tat cag ggt att ggc ccg gca tgg acg aaa gag agt 1152
Leu Thr Pro Tyr Tyr Gln Gly Ile Gly Pro Ala Trp Thr Lys Glu Ser
370 375 380
gtg cta ccg ggt ggc gcc att gaa ggc gac cgc gac gat tat gcc gct 1200
Val Leu Pro Gly Gly Ala Ile Glu Gly Asp Arg Asp Asp Tyr Ala Ala
385 390 395 400
cgc ctg cgc cgc cgc tat ccg ttc ctg act gaa tcg ctg gcg cgt cat 1248
Arg Leu Arg Arg Arg Tyr Pro Phe Leu Thr Glu Ser Leu Ala Arg His
405 410 415
tac gct cgc act tac ggc agc aac agc gag ctg ctg ctc ggc aat gcg 1296
Tyr Ala Arg Thr Tyr Gly Ser Asn Ser Glu Leu Leu Leu Gly Asn Ala
420 425 430
gga acg gta agc gat ctc ggg gaa gat ttc ggt cat gag ttc tac gaa 1344
Gly Thr Val Ser Asp Leu Gly Glu Asp Phe Gly His Glu Phe Tyr Glu
435 440 445
gcg gag ctg aaa tac ctg gtg gat cac gaa tgg gtc cgc cgc gcc gac 1392
Ala Glu Leu Lys Tyr Leu Val Asp His Glu Trp Val Arg Arg Ala Asp
450 455 460
gac gcc ctg tgg cgt cgc aca aaa caa ggc atg tgg cta aat gcg gat 1440
Asp Ala Leu Trp Arg Arg Thr Lys Gln Gly Met Trp Leu Asn Ala Asp
465 470 475 480
caa caa tct cgt gtg agt cag tgg ctg gtg gag tat acg cag cag agg 1488
Gln Gln Ser Arg Val Ser Gln Trp Leu Val Glu Tyr Thr Gln Gln Arg
485 490 495
tta tcg ctg gcg tcg taa 1506
Leu Ser Leu Ala Ser
500
<210>33
<211>501
<212>PRT
<213>大肠杆菌
<400>33
Met Glu Thr Lys Asp Leu Ile Val Ile Gly Gly Gly Ile Asn Gly Ala
1 5 10 15
Gly Ile Ala Ala Asp Ala Ala Gly Arg Gly Leu Ser Val Leu Met Leu
20 25 30
Glu Ala Gln Asp Leu Ala Cys Ala Thr Ser Ser Ala Ser Ser Lys Leu
35 40 45
Ile Hls Gly Gly Leu Arg Tyr Leu Glu His Tyr Glu Phe Arg Leu Val
50 55 60
Ser Glu Ala Leu Ala Glu Arg Glu Val Leu Leu Lys Met Ala Pro His
65 70 75 80
Ile Ala Phe Pro Met Arg Phe Arg Leu Pro His Arg Pro His Leu Arg
85 90 95
Pro Ala Trp Met Ile Arg Ile Gly Leu Phe Met Tyr Asp His Leu Gly
100 105 110
Lys Arg Thr Ser Leu Pro Gly Ser Thr Gly Leu Arg Phe Gly Ala Asn
115 120 125
Ser Val Leu Lys Pro Glu Ile Lys Arg Gly Phe Glu Tyr Ser Asp Cys
130 135 140
Trp Val Asp Asp Ala Arg Leu Val Leu Ala Asn Ala Gln Met Val Val
145 150 155 160
Arg Lys Gly Gly Glu Val Leu Thr Arg Thr Arg Ala Thr Ser Ala Arg
165 170 175
Arg Glu Asn Gly Leu Trp Ile Val Glu Ala Glu Asp Ile Asp Thr Gly
180 185 190
Lys Lys Tyr Ser Trp Gln Ala Arg Gly Leu Val Asn Ala Thr Gly Pro
195 200 205
Trp Val Lys Gln Phe Phe Asp Asp Gly Met His Leu Pro Ser Pro Tyr
210 215 220
Gly Ile Arg Leu Ile Lys Gly Ser His Ile Val Val Pro Arg Val His
225 230 235 240
Thr Gln Lys Gln Ala Tyr Ile Leu Gln Asn Glu Asp Lys Arg Ile Val
245 250 255
Phe Val Ile Pro Trp Met Asp Glu Phe Ser Ile Ile Gly Thr Thr Asp
260 265 270
Val Glu Tyr Lys Gly Asp Pro Lys Ala Val Lys Ile Glu Glu Ser Glu
275 280 285
Ile Asn Tyr Leu Leu Asn Val Tyr Asn Thr His Phe Lys Lys Gln Leu
290 295 300
Ser Arg Asp Asp Ile Val Trp Thr Tyr Ser Gly Val Arg Pro Leu Cys
305 310 315 320
Asp Asp Glu Ser Asp Ser Pro Gln Ala Ile Thr Arg Asp Tyr Thr Leu
325 330 335
Asp Ile His Asp Glu Asn Gly Lys Ala Pro Leu Leu Ser Val Phe Gly
340 345 350
Gly Lys Leu Thr Thr Tyr Arg Lys Leu Ala Glu His Ala Leu Glu Lys
355 360 365
Leu Thr Pro Tyr Tyr Gln Gly Ile Gly Pro Ala Trp Thr Lys Glu Ser
370 375 380
Val Leu Pro Gly Gly Ala Ile Glu Gly Asp Arg Asp Asp Tyr Ala Ala
385 390 395 400
Arg Leu Arg Arg Arg Tyr Pro Phe Leu Thr Glu Ser Leu Ala Arg His
405 410 415
Tyr Ala Arg Thr Tyr Gly Ser Asn Ser Glu Leu Leu Leu Gly Asn Ala
420 425 430
Gly Thr Val Ser Asp Leu Gly Glu Asp Phe Gly His Glu Phe Tyr Glu
435 440 445
Ala Glu Leu Lys Tyr Leu Val Asp His Glu Trp Val Arg Arg Ala Asp
450 455 460
Asp Ala Leu Trp Arg Arg Thr Lys Gln Gly Met Trp Leu Asn Ala Asp
465 470 475 480
Gln Gln Ser Arg Val Ser Gln Trp Leu Val Glu Tyr Thr Gln Gln Arg
485 490 495
Leu Ser Leu Ala Ser
500
<210>34
<211>1071
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1071)
<223>
<400>34
atg aaa aaa ttg atc aat gat gtg caa gac gta ctg gac gaa caa ctg 48
Met Lys Lys Leu Ile Asn Asp Val Gln Asp Val Leu Asp Glu Gln Leu
1 5 10 15
gca gga ctg gcg aaa gcg cat cca tcg ctg aca ctg cat cag gat ccg 96
Ala Gly Leu Ala Lys Ala His Pro Ser Leu Thr Leu His Gln Asp Pro
20 25 30
gtg tat gtc acc cga gct gat gcc cct gtt gca gga aaa gtc gcc ctg 144
Val Tyr Val Thr Arg Ala Asp Ala Pro Val Ala Gly Lys Val Ala Leu
35 40 45
ctg tcg ggt ggc ggc agc gga cac gag ccg atg cac tgt ggt tat atc 192
Leu Ser Gly Gly Gly Ser Gly His Glu Pro Met His Cys Gly Tyr Ile
50 55 60
ggt cag ggg atg ctt tcg ggg gcc tgt ccg ggc gaa att ttc acc tca 240
Gly Gln Gly Met Leu Ser Gly Ala Cys Pro Gly Glu Ile Phc Thr Ser
65 70 75 80
ccg acg ccc gat aaa atc ttt gaa tgc gcc atg caa gtt gat ggc ggc 288
Pro Thr Pro Asp Lys Ile Phe Glu Cys Ala Met Gln Val Asp Gly Gly
85 90 95
gaa ggt gta ctg ttg att atc aaa aat tac acc ggc gat att ctt aac 336
Glu Gly Val Leu Leu Ile Ile Lys Asn Tyr Thr Gly Asp Ile Leu Asn
100 105 110
ttt gaa aca gcg acc gag tta ctg cac gat agc ggc gta aaa gtg acc 384
Phe Glu Thr Ala Thr Glu Leu Leu His Asp Ser Gly Val Lys Val Thr
115 120 125
act gtg gtc att gat gac gac gtt gcg gta aaa gac agt ctt tat act 432
Thr Val Val Ile Asp Asp Asp Val Ala Val Lys Asp Ser Leu Tyr Thr
130 135 140
gcc ggg cga cgc ggc gtt gcc aac acc gta tta att gaa aaa ctc gta 480
Ala Gly Arg Arg Gly Val Ala Asn Thr Val Leu Ile Glu Lys Leu Val
145 150 155 160
ggc gca gcg gcg gag cgt ggc gac tca ctg gac gcc tgt gcg gaa ctg 528
Gly Ala Ala Ala Glu Arg Gly Asp Ser Leu Asp Ala Cys Ala Glu Leu
165 170 175
ggg cgt aag ctg aat aat caa ggc cac tca ata ggt atc gct ctc ggt 576
Gly Arg Lys Leu Asn Asn Gln Gly His Ser Ile Gly Ile Ala Leu Gly
180 185 190
gcc tgt acc gtt cct gcc gcg ggc aaa cct tct ttt acc ctg gcg gat 624
Ala Cys Thr Val Pro Ala Ala Gly Lys Pro Ser Phe Thr Leu Ala Asp
195 200 205
aat gag atg gag ttt ggc gtc ggc att cat ggt gag ccg ggt att gac 672
Asn Glu Met Glu Phe Gly Val Gly Ile His Gly Glu Pro Gly Ile Asp
210 215 220
cgc cgc ccc ttc tct tcc ctt gat caa acc gtc gat gaa atg ttc gac 720
Arg Arg Pro Phe Ser Ser Leu Asp Gln Thr Val Asp Glu Met Phe Asp
225 230 235 240
acc ctg ctg gta aat ggc tca tac cat cgc act ttg cgt ttc tgg gat 768
Thr Leu Leu Val Asn Gly Ser Tyr His Arg Thr Leu Arg Phe Trp Asp
245 250 255
tat caa caa ggc agt tgg cag gaa gaa caa caa acc aaa caa ccg ctc 816
Tyr Gln Gln Gly Ser Trp Gln Glu Glu Gln Gln Thr Lys Gln Pro Leu
260 265 270
cag tct ggc gat cgg gtg att gcg ctg gtt aac aat ctt ggc gca act 864
Gln Ser Gly Asp Arg Val Ile Ala Leu Val Asn Asn Leu Gly Ala Thr
275 280 285
ccg ctt tct gag ctg tac ggc gtc tat aac cgc ctg acc aca cgt tgc 912
Pro Leu Ser Glu Leu Tyr Gly Val Tyr Asn Arg Leu Thr Thr Arg Cys
290 295 300
cag caa gcg gga ttg act atc gaa cgt aat tta att ggc gcg tac tgc 960
Gln Gln Ala Gly Leu Thr Ile Glu Arg Asn Leu Ile Gly Ala Tyr Cys
305 310 315 320
acc tca ctg gat atg acc ggt ttc tca atc acc tta ctg aaa gtt gat 1008
Thr Ser Leu Asp Met Thr Gly Phe Ser Ile Thr Leu Leu Lys Val Asp
325 330 335
gac gaa acg ctg gca ctc tgg gac gcc ccg gtc cac acc ccg gcc ctt 1056
Asp Glu Thr Leu Ala Leu Trp Asp Ala Pro Val His Thr Pro Ala Leu
340 345 350
aac tgg ggt aaa taa 1071
Asn Trp Gly Lys
355
<210>35
<211>356
<212>PRT
<213>大肠杆菌
<400>35
Met Lys Lys Leu Ile Asn Asp Val Gln Asp Val Leu Asp Glu Gln Leu
1 5 10 15
Ala Gly Leu Ala Lys Ala His Pro Ser Leu Thr Leu His Gln Asp Pro
20 25 30
Val Tyr Val Thr Arg Ala Asp Ala Pro Val Ala Gly Lys Val Ala Leu
35 40 45
Leu Ser Gly Gly Gly Ser Gly His Glu Pro Met His Cys Gly Tyr Ile
50 55 60
Gly Gln Gly Met Leu Ser Gly Ala Cys Pro Gly Glu Ile Phe Thr Ser
65 70 75 80
Pro Thr Pro Asp Lys Ile Phe Glu Cys Ala Met Gln Val Asp Gly Gly
85 90 95
Glu Gly Val Leu Leu Ile Ile Lys Asn Tyr Thr Gly Asp Ile Leu Asn
100 105 110
Phe Glu Thr Ala Thr Glu Leu Leu His Asp Ser Gly Val Lys Val Thr
115 120 125
Thr Val Val Ile Asp Asp Asp Val Ala Val Lys Asp Ser Leu Tyr Thr
130 135 140
Ala Gly Arg Arg Gly Val Ala Asn Thr Val Leu Ile Glu Lys Leu Val
145 150 155 160
Gly Ala Ala Ala Glu Arg Gly Asp Ser Leu Asp Ala Cys Ala Glu Leu
165 170 175
Gly Arg Lys Leu Asn Asn Gln Gly His Ser Ile Gly Ile Ala Leu Gly
180 185 190
Ala Cys Thr Val Pro Ala Ala Gly Lys Pro Ser Phe Thr Leu Ala Asp
195 200 205
Asn Glu Met Glu Phe Gly Val Gly Ile His Gly Glu Pro Gly Ile Asp
210 215 220
Arg Arg Pro Phe Ser Ser Leu Asp Gln Thr Val Asp Glu Met Phe Asp
225 230 235 240
Thr Leu Leu Val Asn Gly Ser Tyr His Arg Thr Leu Arg Phe Trp Asp
245 250 255
Tyr Gln Gln Gly Ser Trp Gln Glu Glu Gln Gln Thr Lys Gln Pro Leu
260 265 270
Gln Ser Gly Asp Arg Val Ile Ala Leu Val Asn Asn Leu Gly Ala Thr
275 280 285
Pro Leu Ser Glu Leu Tyr Gly Val Tyr Asn Arg Leu Thr Thr Arg Cys
290 295 300
Gln Gln Ala Gly Leu Thr Ile Glu Arg Asn Leu Ile Gly Ala Tyr Cys
305 310 315 320
Thr Ser Leu Asp Met Thr Gly Phe Ser Ile Thr Leu Leu Lys Val Asp
325 330 335
Asp Glu Thr Leu Ala Leu Trp Asp Ala Pro Val His Thr Pro Ala Leu
340 345 350
Asn Trp Gly Lys
355
<210>36
<211>633
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(633)
<223>
<400>36
atg tca ctg agc aga act caa att gtt aac tgg ctc act cgt tgt ggc 48
Met Ser Leu Ser Arg Thr Gln Ile Val Asn Trp Leu Thr Arg Cys Gly
1 5 10 15
gat att ttc agc acc gag agc gag tat ctt acc gga ctg gat cgc gaa 96
Asp Ile Phe Ser Thr Glu Ser Glu Tyr Leu Thr Gly Leu Asp Arg Glu
20 25 30
att ggc gat gct gac cac ggg cta aat atg aac cga ggc ttt agc aaa 144
Ile Gly Asp Ala Asp His Gly Leu Asn Met Asn Arg Gly Phe Ser Lys
35 40 45
gtg gtg gaa aaa ctc cct gct atc gca gat aaa gat atc ggt ttc att 192
Val Val Glu Lys Leu Pro Ala Ile Ala Asp Lys Asp Ile Gly Phe Ile
50 55 60
ctc aag aat acc ggt atg acg ctg ctt tcc agc gtc ggt ggt gcc agt 240
Leu Lys Asn Thr Gly Met Thr Leu Leu Ser Ser Val Gly Gly Ala Ser
65 70 75 80
ggt ccg ctg ttc ggt acc ttc ttt atc cgc gcc gca cag gcg acc cag 288
Gly Pro Leu Phe Gly Thr Phe Phe Ile Arg Ala Ala Gln Ala Thr Gln
85 90 95
gca cgg caa agc ctg aca ctg gaa gag ctt tat cag atg ttc cgc gat 336
Ala Arg Gln Ser Leu Thr Leu Glu Glu Leu Tyr Gln Met Phe Arg Asp
100 105 110
ggc gcg gac ggc gta atc agt cgc ggg aaa gcc gaa cct ggc gat aaa 384
Gly Ala Asp Gly Val Ile Ser Arg Gly Lys Ala Glu Pro Gly Asp Lys
115 120 125
acc atg tgt gat gtg tgg gtg ccg gtg gtg gaa tcg tta cgt cag tcc 432
Thr Met Cys Asp Val Trp Val Pro Val Val Glu Ser Leu Arg Gln Ser
130 135 140
agc gag caa aat ctc tct gtt ccg gtg gcg ctc gaa gct gcc agt agc 480
Ser Glu Gln Asn Leu Ser Val Pro Val Ala Leu Glu Ala Ala Ser Ser
145 150 155 160
atc gcc gaa tcc gct gca caa agt acg att acg atg caa gcc cgc aaa 528
Ile Ala Glu Ser Ala Ala Gln Ser Thr Ile Thr Met Gln Ala Arg Lys
165 170 175
ggc cgc gcc agt tat ctc ggt gaa cgc agt att ggt cac cag gat ccc 576
Gly Arg Ala Ser Tyr Leu Gly Glu Arg Ser Ile Gly His Gln Asp Pro
180 185 190
ggc gcg acc tcg gtg atg ttt atg atg caa atg ttg gcg tta gcc gca 624
Gly Ala Thr Ser Val Met Phe Met Met Gln Met Leu Ala Leu Ala Ala
195 200 205
aaa gag taa 633
Lys Glu
210
<210>37
<211>210
<212>PRT
<213>大肠杆菌
<400>37
Met Ser Leu Ser Arg Thr Gln Ile Val Asn Trp Leu Thr Arg Cys Gly
1 5 10 15
Asp Ile Phe Ser Thr Glu Ser Glu Tyr Leu Thr Gly Leu Asp Arg Glu
20 25 30
Ile Gly Asp Ala Asp His Gly Leu Asn Met Asn Arg Gly Phe Ser Lys
35 40 45
Val Val Glu Lys Leu Pro Ala Ile Ala Asp Lys Asp Ile Gly Phe Ile
50 55 60
Leu Lys Asn Thr Gly Met Thr Leu Leu Ser Ser Val Gly Gly Ala Ser
65 70 75 80
Gly Pro Leu Phe Gly Thr Phe Phe Ile Arg Ala Ala Gln Ala Thr Gln
85 90 95
Ala Arg Gln Ser Leu Thr Leu Glu Glu Leu Tyr Gln Met Phe Arg Asp
100 105 110
Gly Ala Asp Gly Val Ile Ser Arg Gly Lys Ala Glu Pro Gly Asp Lys
115 120 125
Thr Met Cys Asp Val Trp Val Pro Val Val Glu Ser Leu Arg Gln Ser
130 135 140
Ser Glu Gln Asn Leu Ser Val Pro Val Ala Leu Glu Ala Ala Ser Ser
145 150 155 160
Ile Ala Glu Ser Ala Ala Gln Ser Thr Ile Thr Met Gln Ala Arg Lys
165 170 175
Gly Arg Ala Ser Tyr Leu Gly Glu Arg Ser Ile Gly His Gln Asp Pro
180 185 190
Gly Ala Thr Ser Val Met Phe Met Met Gln Met Leu Ala Leu Ala Ala
195 200 205
Lys Glu
210
<210>38
<211>1419
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1419)
<223>
<400>38
atg gta aac ctg gtc ata gtt tca cat agc agc cga ctg gga gaa ggt 48
Met Val Asn Leu Val Ile Val Ser His Ser Ser Arg Leu Gly Glu Gly
1 5 10 15
gtc ggt gaa tta gcc cgt cag atg tta atg agt gat agt tgt aaa atc 96
Val Gly Glu Leu Ala Arg Gln Met Leu Met Ser Asp Ser Cys Lys Ile
20 25 30
gcc att gcc gcg gga att gac gat cca caa aat ccc att ggt acc gat 144
Ala Ile Ala Ala Gly Ile Asp Asp Pro Gln Asn Pro Ile Gly Thr Asp
35 40 45
gcc gtc aaa gtg atg gag gcc atc gaa tct gtt gct gat gcc gac cat 192
Ala Val Lys Val Met Glu Ala Ile Glu Ser Val Ala Asp Ala Asp His
50 55 60
gtg ctg gtc atg atg gat atg ggt agc gca tta ttg agt gct gaa act 240
Val Leu Val Met Met Asp Met Gly Ser Ala Leu Leu Ser Ala Glu Thr
65 70 75 80
gcg ctg gaa ttg ctg gct ccc gag atc gcc gca aaa gta cgt ttg tgt 288
Ala Leu Glu Leu Leu Ala Pro Glu Ile Ala Ala Lys Val Arg Leu Cys
85 90 95
gct gcg ccg ttg gtc gaa ggt aca ctg gca gca acg gtc agc gcg gcc 336
Ala Ala Pro Leu Val Glu Gly Thr Leu Ala Ala Thr Val Ser Ala Ala
100 105 110
tcg ggg gcg gat atc gac aaa gtt atc ttt gac gcc atg cat gcg ctg 384
Ser Gly Ala Asp Ile Asp Lys Val Ile Phe Asp Ala Met His Ala Leu
115 120 125
gaa gcc aaa cgt gaa caa ctg ggt tta ccg tcc tcc gac act gaa atc 432
Glu Ala Lys Arg Glu Gln Leu Gly Leu Pro Ser Ser Asp Thr Glu Ile
130 135 140
tct gac aca tgt cct gcg tac gat gaa gaa gcc cgt tct ctg gcg gtg 480
Ser Asp Thr Cys Pro Ala Tyr Asp Glu Glu Ala Arg Ser Leu Ala Val
145 150 155 160
gtc ata aaa aac cgt aac ggc ctg cat gta cgt ccg gcc tcc cgg ctg 528
Val Ile Lys Asn Arg Asn Gly Leu His Val Arg Pro Ala Ser Arg Leu
165 170 175
gtt tat acc tta tcg aca ttt aat gcc gat atg ttg ctg gaa aaa aac 576
Val Tyr Thr Leu Ser Thr Phe Asn Ala Asp Met Leu Leu Glu Lys Asn
180 185 190
ggc aaa tgc gtc aca cca gag agt att aac cag att gcg tta cta caa 624
Gly Lys Cys Val Thr Pro Glu Ser Ile Asn Gln Ile Ala Leu Leu Gln
195 200 205
gtt cgc tat aac gat acg ctg cgc ctg att gcg aaa ggg cca gaa gct 672
Val Arg Tyr Asn Asp Thr Leu Arg Leu Ile Ala Lys Gly Pro Glu Ala
210 215 220
gaa gag gca ctg atc gct ttc cgt cag ctg gct gaa gat aac ttt ggt 720
Glu Glu Ala Leu Ile Ala Phe Arg Gln Leu Ala Glu Asp Asn Phe Gly
225 230 235 240
gaa acg gag gaa gtc gct cca cct act ctg cgt ccc gtt ccg cct gtt 768
Glu Thr Glu Glu Val Ala Pro Pro Thr Leu Arg Pro Val Pro Pro Val
245 250 255
tcg ggt aaa gcc ttt tat tat caa cca gtt tta tgt acg gta cag gca 816
Ser Gly Lys Ala Phe Tyr Tyr Gln Pro Val Leu Cys Thr Val Gln Ala
260 265 270
aaa tca acc ctg acc gtg gaa gaa gaa caa gat cga tta cgc cag gct 864
Lys Ser Thr Leu Thr Val Glu Glu Glu Gln Asp Arg Leu Arg Gln Ala
275 280 285
att gac ttc acg tta tta gat ctg atg acg tta aca gcg aaa gca gaa 912
Ile Asp Phe Thr Leu Leu Asp Leu Met Thr Leu Thr Ala Lys Ala Glu
290 295 300
gcc agc ggg ctt gac gat att gcc gca atc ttt tct ggt cac cat aca 960
Ala Ser Gly Leu Asp Asp Ile Ala Ala Ile Phe Ser Gly His His Thr
305 310 315 320
ctg tta gat gat ccg gaa ctg ctg gcg gcg gca agc gaa ctc ctt cag 1008
Leu Leu Asp Asp Pro Glu Leu Leu Ala Ala Ala Ser Glu Leu Leu Gln
325 330 335
cat gaa cat tgc acg gca gaa tat gcc tgg cag caa gtt ctt aaa gaa 1056
His Glu His Cys Thr Ala Glu Tyr Ala Trp Gln Gln Val Leu Lys Glu
340 345 350
ctt agc cag caa tac cag caa ctg gat gat gaa tat cta caa gct cgc 1104
Leu Ser Gln Gln Tyr Gln Gln Leu Asp Asp Glu Tyr Leu Gln Ala Arg
355 360 365
tat att gat gtg gac gat ctt ctg cat cgc acc ctg gtc cac ctg acc 1152
Tyr Ile Asp Val Asp Asp Leu Leu His Arg Thr Leu Val His Leu Thr
370 375 380
caa acg aaa gaa gaa ctc ccg cag ttt aac tcg cca act att cta ctg 1200
Gln Thr Lys Glu Glu Leu Pro Gln Phe Asn Ser Pro Thr Ile Leu Leu
385 390 395 400
gcg gag aac att tat cct tcc aca gta ctg caa ctg gat ccg gcg gtt 1248
Ala Glu Asn Ile Tyr Pro Ser Thr Val Leu Gln Leu Asp Pro Ala Val
405 410 415
gta aaa ggt atc tgc ctt agc gcc gga agt ccg gta tcc cac agc gcc 1296
Val Lys Gly Ile Cys Leu Ser Ala Gly Ser Pro Val Ser His Ser Ala
420 425 430
cta atc gcc cgt gaa ctg ggg att ggc tgg att tgc cag cag ggt gag 1344
Leu Ile Ala Arg Glu Leu Gly Ile Gly Trp Ile Cys Gln Gln Gly Glu
435 440 445
aaa ctg tat gcg ata caa cca gaa gaa acg cta acg ctg gac gtt aaa 1392
Lys Leu Tyr Ala Ile Gln Pro Glu Glu Thr Leu Thr Leu Asp Val Lys
450 455 460
acg caa cgt ttc aac cgt cag ggt taa 1419
Thr Gln Arg Phe Asn Arg Gln Gly
465 470
<210>39
<211>472
<212>PRT
<213>大肠杆菌
<400>39
Met Val Asn Leu Val Ile Val Ser His Ser Ser Arg Leu Gly Glu Gly
1 5 10 15
Val Gly Glu Leu Ala Arg Gln Met Leu Met Ser Asp Ser Cys Lys Ile
20 25 30
Ala Ile Ala Ala Gly Ile Asp Asp Pro Gln Asn Pro Ile Gly Thr Asp
35 40 45
Ala Val Lys Val Met Glu Ala Ile Glu Ser Val Ala Asp Ala Asp His
50 55 60
Val Leu Val Met Met Asp Met Gly Ser Ala Leu Leu Ser Ala Glu Thr
65 70 75 80
Ala Leu Glu Leu Leu Ala Pro Glu Ile Ala Ala Lys Val Arg Leu Cys
85 90 95
Ala Ala Pro Leu Val Glu Gly Thr Leu Ala Ala Thr Val Ser Ala Ala
100 105 110
Ser Gly Ala Asp Ile Asp Lys Val Ile Phe Asp Ala Met His Ala Leu
115 120 125
Glu Ala Lys Arg Glu Gln Leu Gly Leu Pro Ser Ser Asp Thr Glu Ile
130 135 140
Ser Asp Thr Cys Pro Ala Tyr Asp Glu Glu Ala Arg Ser Leu Ala Val
145 150 155 160
Val Ile Lys Asn Arg Asn Gly Leu His Val Arg Pro Ala Ser Arg Leu
165 170 175
Val Tyr Thr Leu Ser Thr Phe Asn Ala Asp Met Leu Leu Glu Lys Asn
180 185 190
Gly Lys Cys Val Thr Pro Glu Ser Ile Asn Gln Ile Ala Leu Leu Gln
195 200 205
Val Arg Tyr Asn Asp Thr Leu Arg Leu Ile Ala Lys Gly Pro Glu Ala
210 215 220
Glu Glu Ala Leu Ile Ala Phe Arg Gln Leu Ala Glu Asp Asn Phe Gly
225 230 235 240
Glu Thr Glu Glu Val Ala Pro Pro Thr Leu Arg Pro Val Pro Pro Val
245 250 255
Ser Gly Lys Ala Phe Tyr Tyr Gln Pro Val Leu Cys Thr Val Gln Ala
260 265 270
Lys Ser Thr Leu Thr Val Glu Glu Glu Gln Asp Arg Leu Arg Gln Ala
275 280 285
Ile Asp Phe Thr Leu Leu Asp Leu Met Thr Leu Thr Ala Lys Ala Glu
290 295 300
Ala Ser Gly Leu Asp Asp Ile Ala Ala Ile Phe Ser Gly His His Thr
305 310 315 320
Leu Leu Asp Asp Pro Glu Leu Leu Ala Ala Ala Ser Glu Leu Leu Gln
325 330 335
His Glu His Cys Thr Ala Glu Tyr Ala Trp Gln Gln Val Leu Lys Glu
340 345 350
Leu Ser Gln Gln Tyr Gln Gln Leu Asp Asp Glu Tyr Leu Gln Ala Arg
355 360 365
Tyr Ile Asp Val Asp Asp Leu Leu His Arg Thr Leu Val His Leu Thr
370 375 380
Gln Thr Lys Glu Glu Leu Pro Gln Phe Asn Ser Pro Thr Ile Leu Leu
385 390 395 400
Ala Glu Asn Ile Tyr Pro Ser Thr Val Leu Gln Leu Asp Pro Ala Val
405 410 415
Val Lys Gly Ile Cys Leu Ser Ala Gly Ser Pro Val Ser His Ser Ala
420 425 430
Leu Ile Ala Arg Glu Leu Gly Ile Gly Trp Ile Cys Gln Gln Gly Glu
435 440 445
Lys Leu Tyr Ala Ile Gln Pro Glu Glu Thr Leu Thr Leu Asp Val Lys
450 455 460
Thr Gln Arg Phe Asn Arg Gln Gly
465 470
<210>40
<211>1776
<212>DNA
<213>粟酒裂殖酵母(Schizosaccharomyces pombe)
<220>
<221>CDS
<222>(1)..(1776)
<223>
<400>40
atg tct gta aaa caa ttt gta tct gaa gga cat ata gta cgc cca tac 48
Met Ser Val Lys Gln Phe Val Ser Glu Gly His Ile Val Arg Pro Tyr
1 5 10 15
ttg ctt gga ctt gct aga agt aac cct ggc cta act gtt att gaa cat 96
Leu Leu Gly Leu Ala Arg Ser Asn Pro Gly Leu Thr Val Ile Glu His
20 25 30
gat cgt gta att tac agg aca gcc tct gcc ccg ggt tct ggt gat ccg 144
Asp Arg Val Ile Tyr Arg Thr Ala Ser Ala Pro Gly Ser Gly Asp Pro
35 40 45
cct aaa gta act tta gtg tct ggt ggt ggt agt ggg cac gag cct aca 192
Pro Lys Val Thr Leu Val Ser Gly Gly Gly Ser Gly His Glu Pro Thr
50 55 60
cat gct ggt ttt gtt ggt gat gga gcc ttg gac gcc gta gct tgt gga 240
His Ala Gly Phe Val Gly Asp Gly Ala Leu Asp Ala Val Ala Cys Gly
65 70 75 80
gac att ttt gct tct ccc tct act aaa caa att tac tct gct ctc aaa 288
Asp Ile Phe Ala Ser Pro Ser Thr Lys Gln Ile Tyr Ser Ala Leu Lys
85 90 95
gct gtt gca tct cct aag gga act tta att att gtc aaa aat tac aca 336
Ala Val Ala Ser Pro Lys Gly Thr Leu Ile Ile Val Lys Asn Tyr Thr
100 105 110
ggt gat att att cat ttt ggc ttg gca gct gag aga gcc aag gca gct 384
Gly Asp Ile Ile His Phe Gly Leu Ala Ala Glu Arg Ala Lys Ala Ala
115 120 125
gga atg aac gtt gag ctt gtt gcc gta gga gat gat gtt tct gta ggc 432
Gly Met Asn Val Glu Leu Val Ala Val Gly Asp Asp Val Ser Val Gly
130 135 140
aaa aag aga ggt gca ctc gta gga cga aga ggt tta gga gcc act gtg 480
Lys Lys Arg Gly Ala Leu Val Gly Arg Arg Gly Leu Gly Ala Thr Val
145 150 155 160
cta gta cac aaa att gcg gga tct gca gca gca tta gga ctg gat ctg 528
Leu Val His Lys Ile Ala Gly Ser Ala Ala Ala Leu Gly Leu Asp Leu
165 170 175
cat caa gtt gct caa gtg gcc caa tca gtg att gac aat gct gct acc 576
His Gln Val Ala Gln Val Ala Gln Ser Val Ile Asp Asn Ala Ala Thr
180 185 190
ata gca gct tca ttg gat cac tgc gcc gtt ccc ggt cgc aaa ttt gaa 624
Ile Ala Ala Ser Leu Asp His Cys Ala Val Pro Gly Arg Lys Phe Glu
195 200 205
aca aac ttg ggt cca gac gag tat gag att gga atg ggt att cat aac 672
Thr Asn Leu Gly Pro Asp Glu Tyr Glu Ile Gly Met Gly Ile His Asn
210 215 220
gag cct gga acc ttc aaa tca tca cca ctt cca tct att ccc gag cta 720
Glu Pro Gly Thr Phe Lys Ser Ser Pro Leu Pro Ser Ile Pro Glu Leu
225 230 235 240
gtt aca gag atg ctt tcg att ttg ttt ggt gag aaa aac ccg gac aat 768
Val Thr Glu Met Leu Ser Ile Leu Phe Gly Glu Lys Asn Pro Asp Asn
245 250 255
agt ttc gta gag ttt tct tca aaa gac gat gtt att ctt cta gta aac 816
Ser Phe Val Glu Phe Ser Ser Lys Asp Asp Val Ile Leu Leu Val Asn
260 265 270
aat atg ggt ggt atg tcc aac tta gag ttg gga tat gct act gaa gta 864
Asn Met Gly Gly Met Ser Asn Leu Glu Leu Gly Tyr Ala Thr Glu Val
275 280 285
gtt tcg gaa caa tta gcc aaa cga ggt atc att cct aaa aga acc atg 912
Val Ser Glu Gln Leu Ala Lys Arg Gly Ile Ile Pro Lys Arg Thr Met
290 295 300
tct ggt act ttt gta acc gca ttg aac gga ccg ggt ttt gga att aca 960
Ser Gly Thr Phe Val Thr Ala Leu Asn Gly Pro Gly Phe Gly Ile Thr
305 310 315 320
ttg gtc aat gct tct aag gct act cca gat att ttc aaa tat ttt gac 1008
Leu Val Asn Ala Ser Lys Ala Thr Pro Asp Ile Phe Lys Tyr Phe Asp
325 330 335
ttg cca aca act gct agt gga tgg aac gtt tct tat cat aac gca aag 1056
Leu Pro Thr Thr Ala Ser Gly Trp Asn Val Ser Tyr His Asn Ala Lys
340 345 350
gac tgg gag gtt ttg gct gac ggc aag gtg cca aca gct ccc gct ttg 1104
Asp Trp Glu Val Leu Ala Asp Gly Lys Val Pro Thr Ala Pro Ala Leu
355 360 365
gag cat acc cgt aat gag aag cac agc ggt gta aag gct gac cca aag 1152
Glu His Thr Arg Asn Glu Lys His Ser Gly Val Lys Ala Asp Pro Lys
370 375 380
atg ttt act aaa att tta aaa gct gcc gtt gac gct atc aat gaa ttt 1200
Met Phe Thr Lys Ile Leu Lys Ala Ala Val Asp Ala Ile Asn Glu Phe
385 390 395 400
gag cca aag aca act tgg tac gat acg att gca gga gat ggt gat tgt 1248
Glu Pro Lys Thr Thr Trp Tyr Asp Thr Ile Ala Gly Asp Gly Asp Cys
405 410 415
gga aca acc ctt gtg aat ggt gga gag gcc atc ata aag gct att aat 1296
Gly Thr Thr Leu Val Asn Gly Gly Glu Ala Ile Ile Lys Ala Ile Asn
420 425 430
gat aaa tca att cga ttg gat gat ggt gta aat ggt att gac gat ttg 1344
Asp Lys Ser Ile Arg Leu Asp Asp Gly Val Asn Gly Ile Asp Asp Leu
435 440 445
gcc tat att gtt gag gat tcg atg gga ggt aca tcg ggt ggt ctt tac 1392
Ala Tyr Ile Val Glu Asp Ser Met Gly Gly Thr Ser Gly Gly Leu Tyr
450 455 460
tcg att tat ttg tct gcg cta gcc aaa gga gtt cgt gag tca ggc gat 1440
Ser Ile Tyr Leu Ser Ala Leu Ala Lys Gly Val Arg Glu Ser Gly Asp
465 470 475 480
tcc gaa tta tcc gtg cat acg ttt gcg ttt gca agc aag tat gca ctt 1488
Ser Glu Leu Ser Val His Thr Phe Ala Phe Ala Ser Lys Tyr Ala Leu
485 490 495
gac gct ctt ttc aaa tac act agg gcc cgt aaa gga ttc cgt act ctg 1536
Asp Ala Leu Phe Lys Tyr Thr Arg Ala Arg Lys Gly Phe Arg Thr Leu
500 505 510
atc gat gct att caa cca ttt gtt gaa act tta aat gaa ggt aag gga 1584
Ile Asp Ala Ile Gln Pro Phe Val Glu Thr Leu Asn Glu Gly Lys Gly
515 520 525
ctt gat gct gct gcg aaa gct gcc acg gaa ggt tct gaa caa act aga 1632
Leu Asp Ala Ala Ala Lys Ala Ala Thr Glu Gly Ser Glu Gln Thr Arg
530 535 540
aaa atg gat gct gtc gtt gga aga gcg tct tat gtt gct aaa gag gaa 1680
Lys Met Asp Ala Val Val Gly Arg Ala Ser Tyr Val Ala Lys Glu Glu
545 550 555 560
ctt cat aag ctt gat agt gag gga ggc tta ccg gat cct ggt gcc ttt 1728
Leu His Lys Leu Asp Ser Glu Gly Gly Leu Pro Asp Pro Gly Ala Phe
565 570 575
gcg tta gcg gcg atc ttg aaa gca att gtt gag gct agt gaa cat taa 1776
Ala Leu Ala Ala Ile Leu Lys Ala Ile Val Glu Ala Ser Glu His
580 585 590
<210>41
<211>591
<212>PRT
<213>粟酒裂殖酵母
<400>41
Met Ser Val Lys Gln Phe Val Ser Glu Gly His Ile Val Arg Pro Tyr
1 5 10 15
Leu Leu Gly Leu Ala Arg Ser Asn Pro Gly Leu Thr Val Ile Glu His
20 25 30
Asp Arg Val Ile Tyr Arg Thr Ala Ser Ala Pro Gly Ser Gly Asp Pro
35 40 45
Pro Lys Val Thr Leu Val Ser Gly Gly Gly Ser Gly His Glu Pro Thr
50 55 60
His Ala Gly Phe Val Gly Asp Gly Ala Leu Asp Ala Val Ala Cys Gly
65 70 75 80
Asp Ile Phe Ala Ser Pro Ser Thr Lys Gln Ile Tyr Ser Ala Leu Lys
85 90 95
Ala Val Ala Ser Pro Lys Gly Thr Leu Ile Ile Val Lys Asn Tyr Thr
100 105 110
Gly AspIle Ile Hi s Phe Gly Leu Ala Ala Glu Arg Ala Lys Ala Ala
115 120 125
Gly Met Asn Val Glu Leu Val Ala Val Gly Asp Asp Val Ser Val Gly
130 135 140
Lys Lys Arg Gly Ala Leu Val Gly Arg Arg Gly Leu Gly Ala Thr Val
145 150 155 160
Leu Val His Lys Ile Ala Gly Ser Ala Ala Ala Leu Gly Leu Asp Leu
165 170 175
His Gln Val Ala Gln Val Ala Gln Ser Val Ile Asp Asn Ala Ala Thr
180 185 190
Ile Ala Ala Ser Leu Asp His Cys Ala Val Pro Gly Arg Lys Phe Glu
195 200 205
Thr Asn Leu Gly Pro Asp Glu Tyr Glu Ile Gly Met Gly Ile His Asn
210 215 220
Glu Pro Gly Thr Phe Lys Ser Ser Pro Leu Pro Ser Ile Pro Glu Leu
225 230 235 240
Val Thr Glu Met Leu Ser Ile Leu Phe Gly Glu Lys Asn Pro Asp Asn
245 250 255
Ser Phe Val Glu Phe Ser Ser Lys Asp Asp Val Ile Leu Leu Val Asn
260 265 270
Asn Met Gly Gly Met Ser Asn Leu Glu Leu Gly Tyr Ala Thr Glu Val
275 280 285
Val Ser Glu Gln Leu Ala Lys Arg Gly Ile Ile Pro Lys Arg Thr Met
290 295 300
Ser Gly Thr Phe Val Thr Ala Leu Asn Gly Pro Gly Phe Gly Ile Thr
305 310 315 320
Leu Val Asn Ala Ser Lys Ala Thr Pro Asp Ile Phe Lys Tyr Phe Asp
325 330 335
Leu Pro Thr Thr Ala Ser Gly Trp Asn Val Ser Tyr His Asn Ala Lys
340 345 350
Asp Trp Glu Val Leu Ala Asp Gly Lys Val Pro Thr Ala Pro Ala Leu
355 360 365
Glu His Thr Arg Asn Glu Lys His Ser Gly Val Lys Ala Asp Pro Lys
370 375 380
Met Phe Thr Lys Ile Leu Lys Ala Ala Val Asp Ala Ile Asn Glu Phe
385 390 395 400
Glu Pro Lys Thr Thr Trp Tyr Asp Thr Ile Ala Gly Asp Gly Asp Cys
405 410 415
Gly Thr Thr Leu Val Asn Gly Gly Glu Ala Ile Ile Lys Ala Ile Asn
420 425 430
Asp Lys Ser Ile Arg Leu Asp Asp Gly Val Asn Gly Ile Asp Asp Leu
435 440 445
Ala Tyr Ile Val Glu Asp Ser Met Gly Gly Thr Ser Gly Gly Leu Tyr
450 455 460
Ser Ile Tyr Leu Ser Ala Leu Ala Lys Gly Val Arg Glu Ser Gly Asp
465 470 475 480
Ser Glu Leu Ser Val His Thr Phe Ala Phe Ala Ser Lys Tyr Ala Leu
485 490 495
Asp Ala Leu Phe Lys Tyr Thr Arg Ala Arg Lys Gly Phe Arg Thr Leu
500 505 510
Ile Asp Ala Ile Gln Pro Phe Val Glu Thr Leu Asn Glu Gly Lys Gly
515 520 525
Leu Asp Ala Ala Ala Lys Ala Ala Thr Glu Gly Ser Glu Gln Thr Arg
530 535 540
Lys Met Asp Ala Val Val Gly Arg Ala Ser Tyr Val Ala Lys Glu Glu
545 550 555 560
Leu His Lys Leu Asp Ser Glu Gly Gly Leu Pro Asp Pro Gly Ala Phe
565 570 575
Ala Leu Ala Ala Ile Leu Lys Ala Ile Val Glu Ala Ser Glu His
580 585 590
<210>42
<211>1830
<212>DNA
<213>Pichia angusta
<220>
<221>CDS
<222>(1)..(1830)
<223>
<400>42
atg tcg agc aaa cac tgg aat tac aag caa gac ctg gtc cat gcg cac 48
Met Ser Ser Lys His Trp Asn Tyr Lys Gln Asp Leu Val His Ala His
1 5 10 15
ctc aaa ggc ctg tgt cat gcc aat cca gac ctc caa ttc atc gaa tcc 96
Leu Lys Gly Leu Cys His Ala Asn Pro Asp Leu Gln Phe Ile Glu Ser
20 25 30
gag cgt gtg gtg atc aac aag cac tcc aag ccc gac aag gtg atg atc 144
Glu Arg Val Val Ile Asn Lys His Ser Lys Pro Asp Lys Val Met Ile
35 40 45
cta tct ggt gga ggg tct ggc cac gag cca ttg cac gct ggc ttt gtt 192
Leu Ser Gly Gly Gly Ser Gly His Glu Pro Leu His Ala Gly Phe Val
50 55 60
ggt gaa ggc tgt ttg gac gtt gga gtg gct ggt ttt gtt ttc gcc tcg 240
Gly Glu Gly Cys Leu Asp Val Gly Val Ala Gly Phe Val Phe Ala Ser
65 70 75 80
ccc tct aca aag cag att gtt tca ggt ttg aag gca aag ccc tca gac 288
Pro Ser Thr Lys Gln Ile Val Ser Gly Leu Lys Ala Lys Pro Ser Asp
85 90 95
aaa ggt acg cta att gtg gtg aaa aat tac acc ggc gac att ctt cac 336
Lys Gly Thr Leu Ile Val Val Lys Asn Tyr Thr Gly Asp Ile Leu His
100 105 110
ttt ggg ctt gct gca gag cgg gcc aag gcc gaa ggc gtc ccc gtg gag 384
Phe Gly Leu Ala Ala Glu Arg Ala Lys Ala Glu Gly Val Pro Val Glu
115 120 125
ctg cta att gtc cag gac gac gtt tct gtg ggc aga acc aag aac gga 432
Leu Leu Ile Val Gln Asp Asp Val Ser Val Gly Arg Thr Lys Asn Gly
130 135 140
atg gtg ggc aga cgc ggt ctg gcc ggt acg agt ctg gtg cac aag att 480
Met Val Gly Arg Arg Gly Leu Ala Gly Thr Ser Leu Val His Lys Ile
145 150 155 160
gtc ggt gcc aag gcc gcc aag gac tcg aac aaa gcc tcg ttg agc gag 528
Val Gly Ala Lys Ala Ala Lys Asp Ser Asn Lys Ala Ser Leu Ser Glu
165 170 175
gtg tac cag ctg ggc gag gcc gtg gtg gcc aat ctg gtg acc atc ggc 576
Val Tyr Gln Leu Gly Glu Ala Val Val Ala Asn Leu Val Thr Ile Gly
180 185 190
gcg tcg ctc gac cac tgc aca att ccg ggt aac aga cac cac gag tcc 624
Ala Ser Leu Asp His Cys Thr Ile Pro Gly Asn Arg His His Glu Ser
195 200 205
gag tcc gac gac gag gac gag cag aaa cat ctg ctc aag gag gac gag 672
Glu Ser Asp Asp Glu Asp Glu Gln Lys His Leu Leu Lys Glu Asp Glu
210 215 220
atc gag gtg ggt atg ggg atc cac aac gag tcg ggc atc aag cgc gtt 720
Ile Glu Val Gly Met Gly Ile His Asn Glu Ser Gly Ile Lys Arg Val
225 230 235 240
tcg cca atc ccg acc atc gac acg ctt gtg gca gac ctg ctc aag tac 768
Ser Pro Ile Pro Thr Ile Asp Thr Leu Val Ala Asp Leu Leu Lys Tyr
245 250 255
ttg ctc gac aag agc gac gag gag aga cac tat gtg gac ttc gac tcg 816
Leu Leu Asp Lys Ser Asp Glu Glu Arg His Tyr Val Asp Phe Asp Ser
260 265 270
tcg gac gag gtt gtg ctg atg atc aac aat ttg ggc ggc acg tcg aac 864
Ser Asp Glu Val Val Leu Met Ile Asn Asn Leu Gly Gly Thr Ser Asn
275 280 285
ctc gag ctg tac gct atc cag aac act gtt gtt gag cag ctg gct acc 912
Leu Glu Leu Tyr Ala Ile Gln Asn Thr Val Val Glu Gln Leu Ala Thr
290 295 300
gac tac aag atc aag ccc gca aga gtg tac acg ggc gcg tac acc acg 960
Asp Tyr Lys Ile Lys Pro Ala Arg Val Tyr Thr Gly Ala Tyr Thr Thr
305 310 315 320
tcg cta gac ggt cct ggt ttt tcc atc acg ttg ctg aac gtg act cgg 1008
Ser Leu Asp Gly Pro Gly Phe Ser Ile Thr Leu Leu Asn Val Thr Arg
325 330 335
gcg ggg ggc aag gag gtt ttc gat tgt ctg gac tac cca acc aag gtt 1056
Ala Gly Gly Lys Glu Val Phe Asp Cys Leu Asp Tyr Pro Thr Lys Val
340 345 350
cct ggg tgg aac tcg tcg tac aca acg gca gaa tgg gcg gcg aaa tcc 1104
Pro Gly Trp Asn Ser Ser Tyr Thr Thr Ala Glu Trp Ala Ala Lys Ser
355 360 365
gag tcg ttc gtc atc gac gct ccg cca gtg agc gac gcg tcg gcg acc 1152
Glu Ser Phe Val Ile Asp Ala Pro Pro Val Ser Asp Ala Ser Ala Thr
370 375 380
tcg aaa gtg cgg ttc tca agc agc aca gtc aag gct gtg ttg gag agc 1200
Ser Lys Val Arg Phe Ser Ser Ser Thr Val Lys Ala Val Leu Glu Ser
385 390 395 400
gga tgc aag aag ttg ctg acc aag gag cca aag atc acg ctg tac gac 1248
Gly Cys Lys Lys Leu Leu Thr Lys Glu Pro Lys Ile Thr Leu Tyr Asp
405 410 415
acg gtt gcg ggc gac ggc gac tgc ggc gag acg ctg gcc aac ggc gcg 1296
Thr Val Ala Gly Asp Gly Asp Cys Gly Glu Thr Leu Ala Asn Gly Ala
420 425 430
cac gcg atc ctg gac ctg ctg gct gcc gac aag ctg gag atc acc gac 1344
His Ala Ile Leu Asp Leu Leu Ala Ala Asp Lys Leu Glu Ile Thr Asp
435 440 445
ggt gtc cgg agt ctg acg cag atc act gac gtc gtc gaa acg gct atg 1392
Gly Val Arg Ser Leu Thr Gln Ile Thr Asp Val Val Glu Thr Ala Met
450 455 460
gga ggc acc tct ggt ggg ctt tac tcg atc ttt atc tct gca ctc gcc 1440
Gly Gly Thr Ser Gly Gly Leu Tyr Ser Ile Phe Ile Ser Ala Leu Ala
465 470 475 480
aag tcg ttg aag gac aga gag ctc cag cag ggc gga tac gag gtg acg 1488
Lys Ser Leu Lys Asp Arg Glu Leu Gln Gln Gly Gly Tyr Glu Val Thr
485 490 495
ccg cag atc ctg gct gca tcg ctc aag gac gcc ctg gag tcg ctg tac 1536
Pro Gln Ile Leu Ala Ala Ser Leu Lys Asp Ala Leu Glu Ser Leu Tyr
500 505 510
aga tac aca cgg gcc cgt gct ggc gac cgg act ctg atc gac gcg ctt 1584
Arg Tyr Thr Arg Ala Arg Ala Gly Asp Arg Thr Leu Ile Asp Ala Leu
515 520 525
gcg ccg ttt gtg gag cag ttt gcg gcc agc aag ggt gac ctc aac cag 1632
Ala Pro Phe Val Glu Gln Phe Ala Ala Ser Lys Gly Asp Leu Asn Gln
530 535 540
gct aac aag gcg tgc cac gag gga gca gag tca acg cga aag ctc aag 1680
Ala Asn Lys Ala Cys His Glu Gly Ala Glu Ser Thr Arg Lys Leu Lys
545 550 555 560
gct aag ttt ggc cgc gcg tcc tac gtc agc gag gag gag ttc aag ccg 1728
Ala Lys Phe Gly Arg Ala Ser Tyr Val Ser Glu Glu Glu Phe Lys Pro
565 570 575
ttt gag gcc gag ggc ggg ctg ccg gat ccc ggc gcc atc ggg ctt gct 1776
Phe Glu Ala Glu Gly Gly Leu Pro Asp Pro Gly Ala Ile Gly Leu Ala
580 585 590
gcg ctg gtc gac ggt ttt gcc gag gcg tac agc aaa ata ggc tcc aac 1824
Ala Leu Val Asp Gly Phe Ala Glu Ala Tyr Ser Lys Ile Gly Ser Asn
595 600 605
ttg tag 1830
Leu
<210>43
<211>609
<212>PRT
<213>Pichia angusta
<400>43
Met Ser Ser Lys His Trp Asn Tyr Lys Gln Asp Leu Val His Ala His
1 5 10 15
Leu Lys Gly Leu Cys His Ala Asn Pro Asp Leu Gln Phe Ile Glu Ser
20 25 30
Glu Arg Val Val Ile Asn Lys His Ser Lys Pro Asp Lys Val Met Ile
35 40 45
Leu Ser Gly Gly Gly Ser Gly His Glu Pro Leu His Ala Gly Phe Val
50 55 60
Gly Glu Gly Cys Leu Asp Val Gly Val Ala Gly Phe Val Phe Ala Ser
65 70 75 80
Pro Ser Thr Lys Gln Ile Val Ser Gly Leu Lys Ala Lys Pro Ser Asp
85 90 95
Lys Gly Thr Leu Ile Val Val Lys Asn Tyr Thr Gly Asp Ile Leu His
100 105 110
Phe Gly Leu Ala Ala Glu Arg Ala Lys Ala Glu Gly Val Pro Val Glu
115 120 125
Leu Leu Ile Val Gln Asp Asp Val Ser Val Gly Arg Thr Lys Asn Gly
130 135 140
Met Val Gly Arg Arg Gly Leu Ala Gly Thr Ser Leu Val His Lys Ile
145 150 155 160
Val Gly Ala Lys Ala Ala Lys Asp Ser Asn Lys Ala Ser Leu Ser Glu
165 170 175
Val Tyr Gln Leu Gly Glu Ala Val Val Ala Asn Leu Val Thr Ile Gly
180 185 190
Ala Ser Leu Asp His Cys Thr Ile Pro Gly Asn Arg His His Glu Ser
195 200 205
Glu Ser Asp Asp Glu Asp Glu Gln Lys His Leu Leu Lys Glu Asp Glu
210 215 220
Ile Glu Val Gly Met Gly Ile His Asn Glu Ser Gly Ile Lys Arg Val
225 230 235 240
Ser Pro Ile Pro Thr Ile Asp Thr Leu Val Ala Asp Leu Leu Lys Tyr
245 250 255
Leu Leu Asp Lys Ser Asp Glu Glu Arg His Tyr Val Asp Phe Asp Ser
260 265 270
Ser Asp Glu Val Val Leu Met Ile Asn Asn Leu Gly Gly Thr Ser Asn
275 280 285
Leu Glu Leu Tyr Ala Ile Gln Asn Thr Val Val Glu Gln Leu Ala Thr
290 295 300
Asp Tyr Lys Ile Lys Pro Ala Arg Val Tyr Thr Gly Ala Tyr Thr Thr
305 310 315 320
Ser Leu Asp Gly Pro Gly Phe Ser Ile Thr Leu Leu Asn Val Thr Arg
325 330 335
Ala Gly Gly Lys Glu Val Phe Asp Cys Leu Asp Tyr Pro Thr Lys Val
340 345 350
Pro Gly Trp Asn Ser Ser Tyr Thr Thr Ala Glu Trp Ala Ala Lys Ser
355 360 365
Glu Ser Phe Val Ile Asp Ala Pro Pro Val Ser Asp Ala Ser Ala Thr
370 375 380
Ser Lys Val Arg Phe Ser Ser Ser Thr Val Lys Ala Val Leu Glu Ser
385 390 395 400
Gly Cys Lys Lys Leu Leu Thr Lys Glu Pro Lys Ile Thr Leu Tyr Asp
405 410 415
Thr Val Ala Gly Asp Gly Asp Cys Gly Glu Thr Leu Ala Asn Gly Ala
420 425 430
His Ala Ile Leu Asp Leu Leu Ala Ala Asp Lys Leu Glu Ile Thr Asp
435 440 445
Gly Val Arg Ser Leu Thr Gln Ile Thr Asp Val Val Glu Thr Ala Met
450 455 460
Gly Gly Thr Ser Gly Gly Leu Tyr Ser Ile Phe Ile Ser Ala Leu Ala
465 470 475 480
Lys Ser Leu Lys Asp Arg Glu Leu Gln Gln Gly Gly Tyr Glu Val Thr
485 490 495
Pro Gln Ile Leu Ala Ala Ser Leu Lys Asp Ala Leu Glu Ser Leu Tyr
500 505 510
Arg Tyr Thr Arg Ala Arg Ala Gly Asp Arg Thr Leu Ile Asp Ala Leu
515 520 525
Ala Pro Phe Val Glu Gln Phe Ala Ala Ser Lys Gly Asp Leu Asn Gln
530 535 540
Ala Asn Lys Ala Cys His Glu Gly Ala Glu Ser Thr Arg Lys Leu Lys
545 550 555 560
Ala Lys Phe Gly Arg Ala Ser Tyr Val Ser Glu Glu Glu Phe Lys Pro
565 570 575
Phe Glu Ala Glu Gly Gly Leu Pro Asp Pro Gly Ala Ile Gly Leu Ala
580 585 590
Ala Leu Val Asp Gly Phe Ala Glu Ala Tyr Ser Lys Ile Gly Ser Asn
595 600 605
Leu
<210>44
<211>1827
<212>DNA
<213>巴斯德毕赤酵母(Pichia pastoris)
<220>
<221>CDS
<222>(1)..(1827)
<223>
<400>44
atg tct agt aaa cat tgg gat tac aag aaa gat ctg gta ctc agc cat 48
Met Ser Ser Lys His Trp Asp Tyr Lys Lys Asp Leu Val Leu Ser His
1 5 10 15
ctg gct gga tta tgt cag tcc aac ccc cat gta agg cta att gag tcc 96
Leu Ala Gly Leu Cys Gln Ser Asn Pro His Val Arg Leu Ile Glu Ser
20 25 30
gaa aga gtc gtc atc tcc gcc gag aac cag gag gac aag att acc ttg 144
Glu Arg Val Val Ile Ser Ala Glu Asn Gln Glu Asp Lys Ile Thr Leu
35 40 45
atc tct ggt gga ggt tca gga cac gag cct cta cat gct ggc ttt gtc 192
Ile Ser Gly Gly Gly Ser Gly His Glu Pro Leu His Ala Gly Phe Val
50 55 60
acc aag gac ggt ctt ttg gat gcc gct gtg gcc ggg ttc atc ttt gcc 240
Thr Lys Asp Gly Leu Leu Asp Ala Ala Val Ala Gly Phe Ile Phe Ala
65 70 75 80
tcg cca tct act aaa cag ata ttt agc gca atc aaa gcc aaa cct tcg 288
Ser Pro Ser Thr Lys Gln Ile Phe Ser Ala Ile Lys Ala Lys Pro Ser
85 90 95
aag aag gga act ttg atc atc gtc aag aac tat aca gga gac att ctt 336
Lys Lys Gly Thr Leu Ile Ile Val Lys Asn Tyr Thr Gly Asp Ile Leu
100 105 110
cac ttc ggt ctt gcc gcc gaa aag gcc aag gcc gaa ggt ctc aac gca 384
His Phe Gly Leu Ala Ala Glu Lys Ala Lys Ala Glu Gly Leu Asn Ala
115 120 125
gaa ctg ctg att gtt cag gac gat gtc tct gtt ggt aaa gcc aag aat 432
Glu Leu Leu Ile Val Gln Asp Asp Val Ser Val Gly Lys Ala Lys Asn
130 135 140
ggc ctg gta ggg cgt aga ggt ttg gct gga act tct tta gtg cac aag 480
Gly Leu Val Gly Arg Arg Gly Leu Ala Gly Thr Ser Leu Val His Lys
145 150 155 160
att ttg ggt gcc aaa gct tat tta caa aag gac aac ctg gaa ttg cat 528
Ile Leu Gly Ala Lys Ala Tyr Leu Gln Lys Asp Asn Leu Glu Leu His
165 170 175
cag ctg gtc act ttc ggt gag aag gtc gtt gcc aac ctg gtt aca att 576
Gln Leu Val Thr Phe Gly Glu Lys Val Val Ala Asn Leu Val Thr Ile
180 185 190
ggt gct tcc ttg gac cat gtc acc att cca gcc aga gca aac aaa cag 624
Gly Ala Ser Leu Asp His Val Thr Ile Pro Ala Arg Ala Asn Lys Gln
195 200 205
gag gaa gat gat tca gat gac gag cac ggc tac gaa gtg cta aag cac 672
Glu Glu Asp Asp Ser Asp Asp Glu His Gly Tyr Glu Val Leu Lys His
210 215 220
gat gaa ttt gaa atc gga atg ggt att cac aac gaa cca ggt atc aag 720
Asp Glu Phe Glu Ile Gly Met Gly Ile His Asn Glu Pro Gly Ile Lys
225 230 235 240
aag tcg tcg cca att cca aca gta gac gaa ttg gtg gct gag cta ctg 768
Lys Ser Ser Pro Ile Pro Thr Val Asp Glu Leu Val Ala Glu Leu Leu
245 250 255
gag tac ctt ctt tcg act acc gat aag gac cga aat tac gtc caa ttt 816
Glu Tyr Leu Leu Ser Thr Thr Asp Lys Asp Arg Asn Tyr Val Gln Phe
260 265 270
gac aag aat gat gaa gtt gtg cta ctg atc aat aac ttg gga gga acc 864
Asp Lys Asn Asp Glu Val Val Leu Leu Ile Asn Asn Leu Gly Gly Thr
275 280 285
tca gtg ttg gaa ttg tac gct att cag aac att gtt gtc gac caa ctg 912
Ser Val Leu Glu Leu Tyr Ala Ile Gln Asn Ile Val Val Asp Gln Leu
290 295 300
gcc tcc aaa tat tcc att aaa cca gtc cgc att ttc act ggc act ttc 960
Ala Ser Lys Tyr Ser Ile Lys Pro Val Arg Ile Phe Thr Gly Thr Phe
305 310 315 320
acc aca tca ctg gat ggg ccc ggt ttt tct atc act tta ttg aat gct 1008
Thr Thr Ser Leu Asp Gly Pro Gly Phe Ser Ile Thr Leu Leu Asn Ala
325 330 335
acc aag aca gga gat aaa gat atc cta aag ttt tta gac cac aag act 1056
Thr Lys Thr Gly Asp Lys Asp Ile Leu Lys Phe Leu Asp His Lys Thr
340 345 350
agt gcc cct ggc tgg aac tcc aac ata tca gac tgg tct ggc agg gtt 1104
Ser Ala Pro Gly Trp Asn Ser Asn Ile Ser Asp Trp Ser Gly Arg Val
355 360 365
gac aat ttt att gtg gct gcc cca gaa ata gac gaa gga gac agt tca 1152
Asp Asn Phe Ile Val Ala Ala Pro Glu Ile Asp Glu Gly Asp Ser Ser
370 375 380
tct aag gta tct gtt gat gct aag ctg tat gct gac ctt ctg gag tct 1200
Ser Lys Val Ser Val Asp Ala Lys Leu Tyr Ala Asp Leu Leu Glu Ser
385 390 395 400
ggt gtg aag aag gtc att tca aaa gag cca aag atc aca ttg tat gac 1248
Gly Val Lys Lys Val Ile Ser Lys Glu Pro Lys Ile Thr Leu Tyr Asp
405 410 415
act gtt gca gga gat ggt gat tgt ggt gaa acg ttg gcg aat ggt tcc 1296
Thr Val Ala Gly Asp Gly Asp Cys Gly Glu Thr Leu Ala Asn Gly Ser
420 425 430
aat gcc att tta aag gct ctg gct gaa gga aag ctt gat ctc aaa gac 1344
Asn Ala Ile Leu Lys Ala Leu Ala Glu Gly Lys Leu Asp Leu Lys Asp
435 440 445
ggt gtt aaa tca ctg gtg caa att aca gac atc gtg gaa act gcc atg 1392
Gly Val Lys Ser Leu Val Gln Ile Thr Asp Ile Val Glu Thr Ala Met
450 455 460
ggt gga aca tct gga ggt ctg tac tcc att ttc atc agc gct tta gca 1440
Gly Gly Thr Ser Gly Gly Leu Tyr Ser Ile Phe Ile Ser Ala Leu Ala
465 470 475 480
aag tct ttg aaa gag aag gaa ctt tcc gag ggt gcc tac aca ctg acc 1488
Lys Ser Leu Lys Glu Lys Glu Leu Ser Glu Gly Ala Tyr Thr Leu Thr
485 490 495
ctt gaa acg atc tct gga tcc tta cag gca gca ctt cag tct cta ttc 1536
Leu Glu Thr Ile Ser Gly Ser Leu Gln Ala Ala Leu Gln Ser Leu Phe
500 505 510
aaa tac aca aga gca cgt act gga gac aga act ctt att gac gct ttg 1584
Lys Tyr Thr Arg Ala Arg Thr Gly Asp Arg Thr Leu Ile Asp Ala Leu
515 520 525
gag cca ttt gta aag gaa ttc gcc aag tct aag gat ctg aaa ctt gca 1632
Glu Pro Phe Val Lys Glu Phe Ala Lys Ser Lys Asp Leu Lys Leu Ala
530 535 540
aac aaa gct gcc cat gat ggt gct gaa gcc aca aga aag ctc gaa gcc 1680
Asn Lys Ala Ala His Asp Gly Ala Glu Ala Thr Arg Lys Leu Glu Ala
545 550 555 560
aag ttt gga aga gca tct tat gtt gcc gaa gaa gag ttc aaa caa ttc 1728
Lys Phe Gly Arg Ala Ser Tyr Val Ala Glu Glu Glu Phe Lys Gln Phe
565 570 575
gaa agt gaa ggt gga cta cct gat cct ggt gct att ggt ctt gcc gct 1776
Glu Ser Glu Gly Gly Leu Pro Asp Pro Gly Ala Ile Gly Leu Ala Ala
580 585 590
ttg att tca gga att act gat gca tac ttc aaa tct gaa acc aag ttg 1824
Leu Ile Ser Gly Ile Thr Asp Ala Tyr Phe Lys Ser Glu Thr Lys Leu
595 600 605
tag 1827
<210>45
<211>608
<212>PRT
<213>巴斯德毕赤酵母
<400>45
Met Ser Ser Lys His Trp Asp Tyr Lys Lys Asp Leu Val Leu Ser His
1 5 10 15
Leu Ala Gly Leu Cys Gln Ser Asn Pro His Val Arg Leu Ile Glu Ser
20 25 30
Glu Arg Val Val Ile Ser Ala Glu Asn Gln Glu Asp Lys Ile Thr Leu
35 40 45
Ile Ser Gly Gly Gly Ser Gly His Glu Pro Leu His Ala Gly Phe Val
50 55 60
Thr Lys Asp Gly Leu Leu Asp Ala Ala Val Ala Gly Phe Ile Phe Ala
65 70 75 80
Ser Pro Ser Thr Lys Gln Ile Phe Ser Ala Ile Lys Ala Lys Pro Ser
85 90 95
Lys Lys Gly Thr Leu Ile Ile Val Lys Asn Tyr Thr Gly Asp Ile Leu
100 105 110
His Phe Gly Leu Ala Ala Glu Lys Ala Lys Ala Glu Gly Leu Asn Ala
115 120 125
Glu Leu Leu Ile Val Gln Asp Asp Val Ser Val Gly Lys Ala Lys Asn
130 135 140
Gly Leu Val Gly Arg Arg Gly Leu Ala Gly Thr Ser Leu Val His Lys
145 150 155 160
Ile Leu Gly Ala Lys Ala Tyr Leu Gln Lys Asp Asn Leu Glu Leu His
165 170 175
Gln Leu Val Thr Phe Gly Glu Lys Val Val Ala Asn Leu Val Thr Ile
180 185 190
Gly Ala Ser Leu Asp His Val Thr Ile Pro Ala Arg Ala Asn Lys Gln
195 200 205
Glu Glu Asp Asp Ser Asp Asp Glu His Gly Tyr Glu Val Leu Lys His
210 215 220
Asp Glu Phe Glu Ile Gly Met Gly Ile His Asn Glu Pro Gly Ile Lys
225 230 235 240
Lys Ser Ser Pro Ile Pro Thr Val Asp Glu Leu Val Ala Glu Leu Leu
245 250 255
Glu Tyr Leu Leu Ser Thr Thr Asp Lys Asp Arg Asn Tyr Va1 Gln Phe
260 265 270
Asp Lys Asn Asp Glu Val Val Leu Leu Ile Asn Asn Leu Gly Gly Thr
275 280 285
Ser Val Leu Glu Leu Tyr Ala Ile Gln Asn Ile Val Val Asp Gln Leu
290 295 300
Ala Ser Lys Tyr Ser Ile Lys Pro Val Arg Ile Phe Thr Gly Thr Phe
305 310 315 320
Thr Thr Ser Leu Asp Gly Pro Gly Phe Ser Ile Thr Leu Leu Asn Ala
325 330 335
Thr Lys Thr Gly Asp Lys Asp Ile Leu Lys Phe Leu Asp His Lys Thr
340 345 350
Ser Ala Pro Gly Trp Asn Ser Asn Ile Ser Asp Trp Ser Gly Arg Val
355 360 365
Asp Asn Phe Ile Val Ala Ala Pro Glu Ile Asp Glu Gly Asp Ser Ser
370 375 380
Ser Lys Val Ser Val Asp Ala Lys Leu Tyr Ala Asp Leu Leu Glu Ser
385 390 395 400
Gly Val Lys Lys Val Ile Ser Lys Glu Pro Lys Ile Thr Leu Tyr Asp
405 410 415
Thr Val Ala Gly Asp Gly Asp Cys Gly Glu Thr Leu Ala Asn Gly Ser
420 425 430
Asn Ala Ile Leu Lys Ala Leu Ala Glu Gly Lys Leu Asp Leu Lys Asp
435 440 445
Gly Val Lys Ser Leu Val Gln Ile Thr Asp Ile Val Glu Thr Ala Met
450 455 460
Gly Gly Thr Ser Gly Gly Leu Tyr Ser Ile Phe Ile Ser Ala Leu Ala
465 470 475 480
Lys Ser Leu Lys Glu Lys Glu Leu Ser Glu Gly Ala Tyr Thr Leu Thr
485 490 495
Leu Glu Thr Ile Ser Gly Ser Leu Gln Ala Ala Leu Gln Ser Leu Phe
500 505 510
Lys Tyr Thr Arg Ala Arg Thr Gly Asp Arg Thr Leu Ile Asp Ala Leu
515 520 525
Glu Pro Phe Val Lys Glu Phe Ala Lys Ser Lys Asp Leu Lys Leu Ala
530 535 540
Asn Lys Ala Ala His Asp Gly Ala Glu Ala Thr Arg Lys Leu Glu Ala
545 550 555 560
Lys Phe Gly Arg Ala Ser Tyr Val Ala Glu Glu Glu Phe Lys Gln Phe
565 570 575
Glu Ser Glu Gly Gly Leu Pro Asp Pro Gly Ala Ile Gly Leu Ala Ala
580 585 590
Leu Ile Ser Gly Ile Thr Asp Ala Tyr Phe Lys Ser Glu Thr Lys Leu
595 600 605
<210>46
<211>1824
<212>DNA
<213>汉逊德巴利酵母(Debaryomyces hansenii)
<220>
<221>CDS
<222>(1)..(1824)
<223>
<400>46
atg tcg tta gct aaa cac tgg gga tac tca aag gat ttg gtt ttg gaa 48
Met Ser Leu Ala Lys His Trp Gly Tyr Ser Lys Asp Leu Val Leu Glu
1 5 10 15
aat tta aag ggt ttg gct gct gca aac ccc aaa att tct ctt ata cca 96
Asn Leu Lys Gly Leu Ala Ala Ala Asn Pro Lys Ile Ser Leu Ile Pro
20 25 30
gca gaa aaa aca gtc gtg tat aat gat tcc tta caa tct aaa gga aac 144
Ala Glu Lys Thr Val Val Tyr Asn Asp Ser Leu Gln Ser Lys Gly Asn
35 40 45
aag aat att aat cgt ata atg gtt att tct gga gga ggt tca ggc cat 192
Lys Asn Ile Asn Arg Ile Met Val Ile Ser Gly Gly Gly Ser Gly His
50 55 60
gaa ccg tta cat gcg gga ttt gtt ggt gta aat gca tta gat gct gct 240
Glu Pro Leu His Ala Gly Phe Val Gly Val Asn Ala Leu Asp Ala Ala
65 70 75 80
gta tct ggt tca ata ttt gcc tcc cct tca gcc aag cag att ttt gca 288
Val Ser Gly Ser Ile Phe Ala Ser Pro Ser Ala Lys Gln Ile Phe Ala
85 90 95
gcc att aag tca att agc tcc aaa caa aac aat tct aaa ggt act ttg 336
Ala Ile Lys Ser Ile Ser Ser Lys Gln Asn Asn Ser Lys Gly Thr Leu
100 105 110
gta att gtc aaa aat tat aca gga gat gtt cta cat ttt gga ctt gct 384
Val Ile Val Lys Asn Tyr Thr Gly Asp Val Leu His Phe Gly Leu Ala
115 120 125
gtc gaa aga gca aaa gca cat ggc tac aaa ata gac atg ata att gtt 432
Val Glu Arg Ala Lys Ala His Gly Tyr Lys Ile Asp Met Ile Ile Val
130 135 140
ggt gat gac gca gct gta ggt agg tca aag ggt gga atg gtt gga aga 480
Gly Asp Asp Ala Ala Val Gly Arg Ser Lys Gly Gly Met Val Gly Arg
145 150 155 160
aga gca ttg gca gcc act gca ttg gtt cac aaa att gtt gga tct gct 528
Arg Ala Leu Ala Ala Thr Ala Leu Val His Lys Ile Val Gly Ser Ala
165 170 175
gct tct gaa att gaa gat cta agc aga ctt aaa ata ttg ggt gat tcc 576
Ala Ser Glu Ile Glu Asp Leu Ser Arg Leu Lys Ile Leu Gly Asp Ser
180 185 190
gtt gcg aat aat acg gtg acc att ggt gcc acc tta gat cat tgc tct 624
Val Ala Asn Asn Thr Val Thr Ile Gly Ala Thr Leu Asp His Cys Ser
195 200 205
gtt cct ggg cgt gat att gca aac ttt gag cct att ggc cag aat gat 672
Val Pro Gly Arg Asp Ile Ala Asn Phe Glu Pro Ile Gly Gln Asn Asp
210 215 220
gct gaa ata ggt cta ggg ata cat aac gaa act tcg gtc aag aag gta 720
Ala Glu Ile Gly Leu Gly Ile His Asn Glu Thr Ser Val Lys Lys Val
225 230 235 240
aac cct gta cca atg ata gac tca tta gtt cag gac tta tta gaa ttc 768
Asn Pro Val Pro Met Ile Asp Ser Leu Val Gln Asp Leu Leu Glu Phe
245 250 255
cta tta aac gaa aat gac aaa gat cgt tac ttt gta cca ttt gat ctc 816
Leu Leu Asn Glu Asn Asp Lys Asp Arg Tyr Phe Val Pro Phe Asp Leu
260 265 270
agt aat gat gaa acg gtc ttg ctt gtc aac aat ctc ggg ggt aca tct 864
Ser Asn Asp Glu Thr Val Leu Leu Val Asn Asn Leu Gly Gly Thr Ser
275 280 285
act tta gaa atg tat gct att aca aat tgc gtt atc gaa aca tta tac 912
Thr Leu Glu Met Tyr Ala Ile Thr Asn Cys Val Ile Glu Thr Leu Tyr
290 295 300
caa caa tac agt ttg aga cca aaa aaa gtg ata gtg gga gaa ttt gct 960
Gln Gln Tyr Ser Leu Arg Pro Lys Lys Val Ile Val Gly Glu Phe Ala
305 310 315 320
acg tct tta aat gcc cct ggg ttt tcg att acc tta ctt aac gtc tct 1008
Thr Ser Leu Asn Ala Pro Gly Phe Ser Ile Thr Leu Leu Asn Val Ser
325 330 335
tgt gca tct aaa caa tcc caa att tcc att tca cat ata atg agt tac 1056
Cys Ala Ser Lys Gln Ser Gln Ile Ser Ile Ser His Ile Met Ser Tyr
340 345 350
ttg gat ttg cca aca gat gcc cct ggt tgg aag gca cat ccg tgt ggg 1104
Leu Asp Leu Pro Thr Asp Ala Pro Gly Trp Lys Ala His Pro Cys Gly
355 360 365
ttt gga ctt gaa aga gac atc aat att gag aca tca atc aat ggt att 1152
Phe Gly Leu Glu Arg Asp Ile Asn Ile Glu Thr Ser Ile Asn Gly Ile
370 375 380
gat tct ttt gtc aag tca caa tta aag ctt tcc aga gaa caa cag acg 1200
Asp Ser Phe Val Lys Ser Gln Leu Lys Leu Ser Arg Glu Gln Gln Thr
385 390 395 400
gac ttt aga agc agt cta gtt aat ggg ttg gaa aaa tta tta gac aaa 1248
Asp Phe Arg Ser Ser Leu Val Asn Gly Leu Glu Lys Leu Leu Asp Lys
405 410 415
gaa cca agc att aca ttt tat gat act gtt gct ggt gat ggt gac tgt 1296
Glu Pro Ser Ile Thr Phe Tyr Asp Thr Val Ala Gly Asp Gly Asp Cys
420 425 430
ggt gaa acc tta gcg tct ggt gca aat gga ata ttg gaa tca tta agg 1344
Gly Glu Thr Leu Ala Ser Gly Ala Asn Gly Ile Leu Glu Ser Leu Arg
435 440 445
aac aac gaa atc tgc ttt gaa gat cca gtt tat tcc ata tct caa ata 1392
Asn Asn Glu Ile Cys Phe Glu Asp Pro Val Tyr Ser Ile Ser Gln Ile
450 455 460
gca aac att gta gag gat aaa atg ggc gga act tca gga ggc tta tat 1440
Ala Asn Ile Val Glu Asp Lys Met Gly Gly Thr Ser Gly Gly Leu Tyr
465 470 475 480
tca att ttc tta acc tcg ttg ata aaa cac ctt caa gat tgt act aca 1488
Ser Ile Phe Leu Thr Ser Leu Ile Lys His Leu Gln Asp Cys Thr Thr
485 490 495
ttg aac tta tgt gaa atg ttt gct agt tct ttg cat aat gcg cta tat 1536
Leu Asn Leu Cys Glu Met Phe Ala Ser Ser Leu His Asn Ala Leu Tyr
500 505 510
cag ggc tta tat aaa tac act agg gca cga gtg ggt gga aga act ttg 1584
Gln Gly Leu Tyr Lys Tyr Thr Arg Ala Arg Val Gly Gly Arg Thr Leu
515 520 525
att gat gct tta gag ccg ttt gtg aat acc ttc aac gac act cta aat 1632
Ile Asp Ala Leu Glu Pro Phe Val Asn Thr Phe Asn Asp Thr Leu Asn
530 535 540
ttc tca aag gct gcc cag gct gct atc gat gga tct gaa tcg act agg 1680
Phe Ser Lys Ala Ala Gln Ala Ala Ile Asp Gly Ser Glu Ser Thr Arg
545 550 555 560
aaa tta gct gca aaa ttt ggg aga gcg tct tat gtc aat gaa caa gaa 1728
Lys Leu Ala Ala Lys Phe Gly Arg Ala Ser Tyr Val Asn Glu Gln Glu
565 570 575
ttt aaa cag ttc gac gaa gaa gga ggc tta ccg gat ccg gga gct att 1776
Phe Lys Gln Phe Asp Glu Glu Gly Gly Leu Pro Asp Pro Gly Ala Ile
580 585 590
gga tta gcc acc tta att gct ggg ttt gcc ggt gtt gac tat aat taa 1824
Gly Leu Ala Thr Leu Ile Ala Gly Phe Ala Gly Val Asp Tyr Asn
595 600 605
<210>47
<211>607
<212>PRT
<213>汉逊德巴利酵母
<400>47
Met Ser Leu Ala Lys His Trp Gly Tyr Ser Lys Asp Leu Val Leu Glu
1 5 10 15
Asn Leu Lys Gly Leu Ala Ala Ala Asn Pro Lys Ile Ser Leu Ile Pro
20 25 30
Ala Glu Lys Thr Val Val Tyr Asn Asp Ser Leu Gln Ser Lys Gly Asn
35 40 45
Lys Asn Ile Asn Arg Ile Met Val Ile Ser Gly Gly Gly Ser Gly His
50 55 60
Glu Pro Leu His Ala Gly Phe Val Gly Val Asn Ala Leu Asp Ala Ala
65 70 75 80
Val Ser Gly Ser Ile Phe Ala Ser Pro Ser Ala Lys Gln Ile Phe Ala
85 90 95
Ala Ile Lys Ser Ile Ser Ser Lys Gln Asn Asn Ser Lys Gly Thr Leu
100 105 110
Val Ile Val Lys Asn Tyr Thr Gly Asp Val Leu His Phe Gly Leu Ala
115 120 125
Val Glu Arg Ala Lys Ala His Gly Tyr Lys Ile Asp Met Ile Ile Val
130 135 140
Gly Asp Asp Ala Ala Val Gly Arg Ser Lys Gly Gly Met Val Gly Arg
145 150 155 160
Arg Ala Leu Ala Ala Thr Ala Leu Val His Lys Ile Val Gly Ser Ala
165 170 175
Ala Ser Glu Ile Glu Asp Leu Ser Arg Leu Lys Ile Leu Gly Asp Ser
180 185 190
Val Ala Asn Asn Thr Val Thr Ile Gly Ala Thr Leu Asp His Cys Ser
195 200 205
Val Pro Gly Arg Asp Ile Ala Asn Phe Glu Pro Ile Gly Gln Asn Asp
210 215 220
Ala Glu Ile Gly Leu Gly Ile His Asn Glu Thr Ser Val Lys Lys Val
225 230 235 240
Asn Pro Val Pro Met Ile Asp Ser Leu Val Gln Asp Leu Leu Glu Phe
245 250 255
Leu Leu Asn Glu Asn Asp Lys Asp Arg Tyr Phe Val Pro Phe Asp Leu
260 265 270
Ser Asn Asp Glu Thr Val Leu Leu Val Asn Asn Leu Gly Gly Thr Ser
275 280 285
Thr Leu Glu Met Tyr Ala Ile Thr Asn Cys Val Ile Glu Thr Leu Tyr
290 295 300
Gln Gln Tyr Ser Leu Arg Pro Lys Lys Val Ile Val Gly Glu Phe Ala
305 310 315 320
Thr Ser Leu Asn Ala Pro Gly Phe Ser Ile Thr Leu Leu Asn Val Ser
325 330 335
Cys Ala Ser Lys Gln Ser Gln Ile Ser Ile Ser His Ile Met Ser Tyr
340 345 350
Leu Asp Leu Pro Thr Asp Ala Pro Gly Trp Lys Ala His Pro Cys Gly
355 360 365
Phe Gly Leu Glu Arg Asp Ile Asn Ile Glu Thr Ser Ile Asn Gly Ile
370 375 380
Asp Ser Phe Val Lys Ser Gln Leu Lys Leu Ser Arg Glu Gln Gln Thr
385 390 395 400
Asp Phe Arg Ser Ser Leu Val Asn Gly Leu Glu Lys Leu Leu Asp Lys
405 410 415
Glu Pro Ser Ile Thr Phe Tyr Asp Thr yal Ala Gly Asp Gly Asp Cys
420 425 430
Gly Glu Thr Leu Ala Ser Gly Ala Asn Gly Ile Leu Glu Ser Leu Arg
435 440 445
Asn Asn Glu Ile Cys Phe Glu Asp Pro Val Tyr Ser Ile Ser Gln Ile
450 455 460
Ala Asn Ile Val Glu Asp Lys Met Gly Gly Thr Ser Gly Gly Leu Tyr
465 470 475 480
Ser Ile Phe Leu Thr Ser Leu Ile Lys His Leu Gln Asp Cys Thr Thr
485 490 495
Leu Asn Leu Cys Glu Met Phe Ala Ser Ser Leu His Asn Ala Leu Tyr
500 505 510
Gln Gly Leu Tyr Lys Tyr Thr Arg Ala Arg Val Gly Gly Arg Thr Leu
515 520 525
Ile Asp Ala Leu Glu Pro Phe Val Asn Thr Phe Asn Asp Thr Leu Asn
530 535 540
Phe Ser Lys Ala Ala Gln Ala Ala Ile Asp Gly Ser Glu Ser Thr Arg
545 550 555 560
Lys Leu Ala Ala Lys Phe Gly Arg Ala Ser Tyr Val Asn Glu Gln Glu
565 570 575
Phe Lys Gln Phe Asp Glu Glu Gly Gly Leu Pro Asp Pro Gly Ala Ile
580 585 590
Gly Leu Ala Thr Leu Ile Ala Gly Phe Ala Gly Val Asp Tyr Asn
595 600 605
<210>48
<211>1752
<212>DNA
<213>蟑螂埃希氏菌(Escherichia blattae)
<220>
<221>CDS
<222>(1)..(1752)
<223>
<400>48
atg gtc tgc ccg cca gac ctt aac tca caa cct ttt act tcc cga cac 48
Met Val Cys Pro Pro Asp Leu Asn Ser Gln Pro Phe Thr Ser Arg His
1 5 10 15
acg cgc tac agc ccg ctg tac ggg tgt gtt gta cct aac gat cag gaa 96
Thr Arg Tyr Ser Pro Leu Tyr Gly Cys Val Val Pro Asn Asp Gln Glu
20 25 30
acc gtt atg tcg caa ttt ttt tat aac caa cgc gaa aat ctt gtc agt 144
Thr Val Met Ser Gln Phe Phe Tyr Asn Gln Arg Glu Asn Leu Val Ser
35 40 45
gac gcc atc gaa ggg gca atg att gcc agc ccg tgg aac aac ctg gcc 192
Asp Ala Ile Glu Gly Ala Met Ile Ala Ser Pro Trp Asn Asn Leu Ala
50 55 60
cgt ctg gag agc gat ccc gcc atc cgc atc gtg gtg cgc cgg gat ctg 240
Arg Leu Glu Ser Asp Pro Ala Ile Arg Ile Val Val Arg Arg Asp Leu
65 70 75 80
gat aaa tcc cgg gtg gcg gtg atc tcc ggg ggc ggt gcc ggt cac gaa 288
Asp Lys Ser Arg Val Ala Val Ile Ser Gly Gly Gly Ala Gly His Glu
85 90 95
ccg gcc cat gtg ggc ttt gtg ggt aaa ggc atg ctg acc gcc gcc gtg 336
Pro Ala His Val Gly Phe Val Gly Lys Gly Met Leu Thr Ala Ala Val
100 105 110
tgt ggt gat ctg ttt gcc tca ccg agc gtg gat gcg gtg ctt acc gcc 384
Cys Gly Asp Leu Phe Ala Ser Pro Ser Val Asp Ala Val Leu Thr Ala
115 120 125
att cag gcg gtc acc ggc gag gcg ggc tgc ctg ctg att gtc aaa aac 432
Ile Gln Ala Val Thr Gly Glu Ala Gly Cys Leu Leu Ile Val Lys Asn
130 135 140
tac acc ggt gac cgg ctg aac ttc ggg ctg gca gca gag aaa gcc cgg 480
Tyr Thr Gly Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Lys Ala Arg
145 150 155 160
cgc atg ggc tac aaa gtg gac atg gtg att gtg ggg gat gat atc tca 528
Arg Met Gly Tyr Lys Val Asp Met Val Ile Val Gly Asp Asp Ile Ser
165 170 175
ctg ccg gag aac aaa cac ccg cgc ggc att gcc ggg acg att atg atc 576
Leu Pro Glu Asn Lys His Pro Arg Gly Ile Ala Gly Thr Ile Met Ile
180 185 190
cac aaa gtg gcg ggg tac ttc gcc gaa acc ggc tgc aac ctg gac acg 624
His Lys Val Ala Gly Tyr Phe Ala Glu Thr Gly Cys Asn Leu Asp Thr
195 200 205
gtc gcc cgg gaa gcc cgg ctg gca atg gag cgg gta ttc agt att ggc 672
Val Ala Arg Glu Ala Arg Leu Ala Met Glu Arg Val Phe Ser Ile Gly
210 215 220
gtg gcc ctt tcc agc tgc cac tta ccg gcc gat ccg cag gat ggc gtg 720
Val Ala Leu Ser Ser Cys His Leu Pro Ala Asp Pro Gln Asp Gly Val
225 230 235 240
cgc cat cac ccg ggc cag gct gag ctg ggc atg ggg atc cac ggg gag 768
Arg His His Pro Gly Gln Ala Glu Leu Gly Met Gly Ile His Gly Glu
245 250 255
ccc ggc gca agc gtc atc gac acc cag aac agc acc gac att gtg cgc 816
Pro Gly Ala Ser Val Ile Asp Thr Gln Asn Ser Thr Asp Ile Val Arg
260 265 270
ctg atg gtg gca aaa atc cgc gct gcc ctg cct gaa acc ggc cgc ctg 864
Leu Met Val Ala Lys Ile Arg Ala Ala Leu Pro Glu Thr Gly Arg Leu
275 280 285
ctg ctg atg ctg aat aac ctt ggc ggc gtc tca gtc acc gaa atg gcg 912
Leu Leu Met Leu Asn Asn Leu Gly Gly Val Ser Val Thr Glu Met Ala
290 295 300
atc ctc acc cgg gag ctg gcc cac tgt gag ctt gcc acc cgc acc gac 960
Ile Leu Thr Arg Glu Leu Ala His Cys Glu Leu Ala Thr Arg Thr Asp
305 310 315 320
tgg ttg atg ggc ccg gca ccg ctg gtc agc gcc ctg gac atg aaa ggc 1008
Trp Leu Met Gly Pro Ala Pro Leu Val Ser Ala Leu Asp Met Lys Gly
325 330 335
ttt tcc atc acc gcc ctg gtc atg gaa gag agc att gaa aaa gcc ctg 1056
Phe Ser Ile Thr Ala Leu Val Met Glu Glu Ser Ile Glu Lys Ala Leu
340 345 350
ctg gca gac gtg gaa acc gca ggc tgg ctg ccg cca gtg cgc ctg cgc 1104
Leu Ala Asp Val Glu Thr Ala Gly Trp Leu Pro Pro Val Arg Leu Arg
355 360 365
gcc agc cag acc cag ccc tgc aat atc cgc agt gcc cgg gtg gca ttc 1152
Ala Ser Gln Thr Gln Pro Cys Asn Ile Arg Ser Ala Arg Val Ala Phe
370 375 380
acc ccg tca gac aac ccg gta gtg ggc cag tat gtg gag acc gtc acc 1200
Thr Pro Ser Asp Asn Pro Val Val Gly Gln Tyr Val Glu Thr Val Thr
385 390 395 400
gcc aca ctc agc gcc cag gaa gcg gaa ctt aac gcc ctg gat gcc aaa 1248
Ala Thr Leu Ser Ala Gln Glu Ala Glu Leu Asn Ala Leu Asp Ala Lys
405 410 415
gtg ggc gac ggc gat acc ggc tcc acc ttt gcc gcc ggg gcc cgc gcc 1296
Val Gly Asp Gly Asp Thr Gly Ser Thr Phe Ala Ala Gly Ala Arg Ala
420 425 430
att gca gaa ctg ctg cac cag cac cag ctg ccg ctc agc cag ctg gat 1344
Ile Ala Glu Leu Leu His Gln His Gln Leu Pro Leu Ser Gln Leu Asp
435 440 445
acc ctg tgc gcc ctg atc ggc gaa cgc tta aca gta gtg atg ggg ggc 1392
Thr Leu Cys Ala Leu Ile Gly Glu Arg Leu Thr Val Val Met Gly Gly
450 455 460
tcc agc ggt gtg ctg atg tcc atc ttc ttt acg gcc gca ggc cag gca 1440
Ser Ser Gly Val Leu Met Ser Ile Phe Phe Thr Ala Ala Gly Gln Ala
465 470 475 480
atc agc gaa ggg aaa ccg gtg gtg gcc gca ctc cag gcg ggc ctg gcg 1488
Ile Ser Glu Gly Lys Pro Val Val Ala Ala Leu Gln Ala Gly Leu Ala
485 490 495
cag atg aag tat tac ggc ggt gcc gat ctg ggt gac cgc acc ctg att 1536
Gln Met Lys Tyr Tyr Gly Gly Ala Asp Leu Gly Asp Arg Thr Leu Ile
500 505 510
gat gcc ctg caa cct gcg ctg gca gct ctg gca cag cat ccg ggg gat 1584
Asp Ala Leu Gln Pro Ala Leu Ala Ala Leu Ala Gln His Pro Gly Asp
515 520 525
ctg gcg gcg gca tac cag gca gcc cgg gac ggg gca gac gcc acc acc 1632
Leu Ala Ala Ala Tyr Gln Ala Ala Arg Asp Gly Ala Asp Ala Thr Thr
530 535 540
cgc gcc acc aaa gcc aat gcc ggt cgc gcc tct tac ctg aac agc gac 1680
Arg Ala Thr Lys Ala Asn Ala Gly Arg Ala Ser Tyr Leu Asn Ser Asp
545 550 555 560
agc ctg gcg ggc aat atg gac ccg ggc gct cac gca gtc gca atg gtc 1728
Ser Leu Ala Gly Asn Met Asp Pro Gly Ala His Ala Val Ala Met Val
565 570 575
ttt aag gcg ctg gcg ggc gcc tga 1752
Phe Lys Ala Leu Ala Gly Ala
580
<210>49
<211>583
<212>PRT
<213>蟑螂埃希氏菌
<400>49
Met Val Cys Pro Pro Asp Leu Asn Ser Gln Pro Phe Thr Ser Arg His
1 5 10 15
Thr Arg Tyr Ser Pro Leu Tyr Gly Cys Val Val Pro Asn Asp Gln Glu
20 25 30
Thr Val Met Ser Gln Phe Phe Tyr Asn Gln Arg Glu Asn Leu Val Ser
35 40 45
Asp Ala Ile Glu Gly Ala Met Ile Ala Ser Pro Trp Asn Asn Leu Ala
50 55 60
Arg Leu Glu Ser Asp Pro Ala Ile Arg Ile Val Val Arg Arg Asp Leu
65 70 75 80
Asp Lys Ser Arg Val Ala Val Ile Ser Gly Gly Gly Ala Gly His Glu
85 90 95
Pro Ala His Val Gly Phe Val Gly Lys Gly Met Leu Thr Ala Ala Val
100 105 110
Cys Gly Asp Leu Phe Ala Ser Pro Ser Val Asp Ala Val Leu Thr Ala
115 120 125
Ile Gln Ala Val Thr Gly Glu Ala Gly Cys Leu Leu Ile Val Lys Asn
130 135 140
Tyr Thr Gly Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Lys Ala Arg
145 150 155 160
Arg Met Gly Tyr Lys Val Asp Met Val Ile Val Gly Asp Asp Ile Ser
165 170 175
Leu Pro Glu Asn Lys His Pro Arg Gly Ile Ala Gly Thr Ile Met Ile
180 185 190
His Lys Val Ala Gly Tyr Phe Ala Glu Thr Gly Cys Asn Leu Asp Thr
195 200 205
Val Ala Arg Glu Ala Arg Leu Ala Met Glu Arg Val Phe Ser Ile Gly
210 215 220
Val Ala Leu Ser Ser Cys His Leu Pro Ala Asp Pro Gln Asp Gly Val
225 230 235 240
Arg His His Pro Gly Gln Ala Glu Leu Gly Met Gly Ile His Gly Glu
245 250 255
Pro Gly Ala Ser Val Ile Asp Thr Gln Asn Ser Thr Asp Ile Val Arg
260 265 270
Leu Met Val Ala Lys Ile Arg Ala Ala Leu Pro Glu Thr Gly Arg Leu
275 280 285
Leu Leu Met Leu Asn Asn Leu Gly Gly Val Ser Val Thr Glu Met Ala
290 295 300
Ile Leu Thr Arg Glu Leu Ala His Cys Glu Leu Ala Thr Arg Thr Asp
305 310 315 320
Trp Leu Met Gly Pro Ala Pro Leu Val Ser Ala Leu Asp Met Lys Gly
325 330 335
Phe Ser Ile Thr Ala Leu Val Met Glu Glu Ser Ile Glu Lys Ala Leu
340 345 350
Leu Ala Asp Val Glu Thr Ala Gly Trp Leu Pro Pro Val Arg Leu Arg
355 360 365
Ala Ser Gln Thr Gln Pro Cys Asn Ile Arg Ser Ala Arg Val Ala Phe
370 375 380
Thr Pro Ser Asp Asn Pro Val Val Gly Gln Tyr Val Glu Thr Val Thr
385 390 395 400
Ala Thr Leu Ser Ala Gln Glu Ala Glu Leu Asn Ala Leu Asp Ala Lys
405 410 415
Val Gly Asp Gly Asp Thr Gly Ser Thr Phe Ala Ala Gly Ala Arg Ala
420 425 430
Ile Ala Glu Leu Leu His Gln His Gln Leu Pro Leu Ser Gln Leu Asp
435 440 445
Thr Leu Cys Ala Leu Ile Gly Glu Arg Leu Thr Val Val Met Gly Gly
450 455 460
Ser Ser Gly Val Leu Met Ser lle Phe Phe Thr Ala Ala Gly Gln Ala
465 470 475 480
Ile Ser Glu Gly Lys Pro Val Val Ala Ala Leu Gln Ala Gly Leu Ala
485 490 495
Gln Met Lys Tyr Tyr Gly Gly Ala Asp Leu Gly Asp Arg Thr Leu Ile
500 505 510
Asp Ala Leu Gln Pro Ala Leu Ala Ala Leu Ala Gln His Pro Gly Asp
515 520 525
Leu Ala Ala Ala Tyr Gln Ala Ala Arg Asp Gly Ala Asp Ala Thr Thr
530 535 540
Arg Ala Thr Lys Ala Asn Ala Gly Arg Ala Ser Tyr Leu Asn Ser Asp
545 550 555 560
Ser Leu Ala Gly Asn Met Asp Pro Gly Ala His Ala Val Ala Met Val
565 570 575
Phe Lys Ala Leu Ala Gly Ala
580
<210>50
<211>1647
<212>DNA
<213>肠杆菌属菌种638
<220>
<22l>CDS
<222>(1)..(1647)
<223>
<400>50
atg tcc aga ttc ttt ttt aat gac cgc aaa cag ctg gtc aac gac gcc 48
Met Ser Arg Phe Phe Phe Asn Asp Arg Lys Gln Leu Val Asn Asp Ala
1 5 10 15
att gaa ggc ata ctg att tcc gcg ccg cac ggg aat ctt gtc aaa ctt 96
Ile Glu Gly Ile Leu Ile Ser Ala Pro His Gly Asn Leu Val Lys Leu
20 25 30
gat atc gat ccg gcc att cgg gtg gtt gcg cgt agc gac tgg gat aaa 144
Asp Ile Asp Pro Ala Ile Arg Val Val Ala Arg Ser Asp Trp Asp Lys
35 40 45
agc cgc gta gcg gtg att tcc ggt ggt ggg tcg ggg cac gaa ccc gct 192
Ser Arg Val Ala Val Ile Ser Gly Gly Gly Ser Gly His Glu Pro Ala
50 55 60
cat gcc gga ttt gtc ggc aaa ggg atg ttg acc gca gcc gtc tgt ggc 240
His Ala Gly Phe Val Gly Lys Gly Met Leu Thr Ala Ala Val Cys Gly
65 70 75 80
gat ctg ttt gcc tca ccg agc gta gat gcg gtg tta aac gcg att gtg 288
Asp Leu Phe Ala Ser Pro Ser Val Asp Ala Val Leu Asn Ala Ile Val
85 90 95
gcg gta acg ggc gat cgc ggt tgc ctg tta atc gtc aaa aat tat acc 336
Ala Val Thr Gly Asp Arg Gly Cys Leu Leu Ile Val Lys Asn Tyr Thr
100 105 110
ggc gat cgg ctt aac ttt ggc ctc gcg gcg gaa aag gcc aaa cgc tat 384
Gly Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Lys Ala Lys Arg Tyr
115 120 125
ggg ctg aag gtt gag atg gtg att gtt gct gat gac atc gcc ctg ccg 432
Gly Leu Lys Val Glu Met Val Ile Val Ala Asp Asp Ile Ala Leu Pro
130 135 140
gat aac aaa cag ccg cgt ggc att gcg ggt acg gcg ctg gta cac aaa 480
Asp Asn Lys Gln Pro Arg Gly Ile Ala Gly Thr Ala Leu Val His Lys
145 150 155 160
att gcc gga tat gca gcc gaa cag ggg aaa tca ctg gct gac gtg cgg 528
Ile Ala Gly Tyr Ala Ala Glu Gln Gly Lys Ser Leu Ala Asp Val Arg
165 170 175
gat att gcg cag cag gcc tgt gac aat atc tgg agc ctg ggc gtg gcg 576
Asp Ile Ala Gln Gln Ala Cys Asp Asn Ile Trp Ser Leu Gly Val Ala
180 185 190
atg caa acg tgc aac ctg ccg ggc agc gac gat gaa gaa ggg cgt atc 624
Met Gln Thr Cys Asn Leu Pro Gly Ser Asp Asp Glu Glu Gly Arg Ile
195 200 205
aag gat gga cat gtc gaa ctg ggg ctg ggc att cac ggc gag ccg ggc 672
Lys Asp Gly His Val Glu Leu Gly Leu Gly Ile His Gly Glu Pro Gly
210 215 220
gcg tcg gtg gtt gat acg cac aac agc aaa gag att atc gac acc ctg 720
Ala Ser Val Val Asp Thr His Asn Ser Lys Glu Ile Ile Asp Thr Leu
225 230 235 240
gtg aag ccg tta aaa gag acg gcc ggc gaa ggc aaa ttt gcg gtg ctg 768
Val Lys Pro Leu Lys Glu Thr Ala Gly Glu Gly Lys Phe Ala Val Leu
245 250 255
att aac aat ctc ggc ggt gta tcg gcg ctg gag atg gcg ctg ctc acg 816
Ile Asn Asn Leu Gly Gly Val Ser Ala Leu Glu Met Ala Leu Leu Thr
260 265 270
aaa gaa ctg gcg gat tct gcg ctg aaa gaa aat att gcg tat ctg att 864
Lys Glu Leu Ala Asp Ser Ala Leu Lys Glu Asn Ile Ala Tyr Leu Ile
275 280 285
ggc cct gcg ccg ctg gta agc tcg ctg gat atg aaa ggc ttt tcg ctg 912
Gly Pro Ala Pro Leu Val Ser Ser Leu Asp Met Lys Gly Phe Ser Leu
290 295 300
tca ctg tta cag ctt aac gat acc ttt gag aaa gcc att aac gca ccc 960
Ser Leu Leu Gln Leu Asn Asp Thr Phe Glu Lys Ala Ile Asn Ala Pro
305 310 315 320
gtc gaa act atc ggc tgg caa aag ccg gta gca ttc gcg cca tta cgc 1008
Val Glu Thr Ile Gly Trp Gln Lys Pro Val Ala Phe Ala Pro Leu Arg
325 330 335
acg ctt tcg cat act gcg att cag gat cgt gtt gaa ttt acg cct tcc 1056
Thr Leu Ser His Thr Ala Ile Gln Asp Arg Val Glu Phe Thr Pro Ser
340 345 350
ggg aac gac gag gtc gca gcg cga gtg gca gcg gcg acg caa acg ttg 1104
Gly Asn Asp Glu Val Ala Ala Arg Val Ala Ala Ala Thr Gln Thr Leu
355 360 365
ctc gct ctg gag aac cgt tta aat gcg ctg gac gcc aaa gtg ggc gac 1152
Leu Ala Leu Glu Asn Arg Leu Asn Ala Leu Asp Ala Lys Val Gly Asp
370 375 380
ggc gat acc ggg tcg act ttt gcg caa ggc gcg cgg gaa att gcg cag 1200
Gly Asp Thr Gly Ser Thr Phe Ala Gln Gly Ala Arg Glu Ile Ala Gln
385 390 395 400
ctt ctg gag caa aaa cag ctt ccg cta aac gat ctt tct aag ctg ctg 1248
Leu Leu Glu Gln Lys Gln Leu Pro Leu Asn Asp Leu Ser Lys Leu Leu
405 410 415
ttg ttg atc ggc gaa cgg ctg gcg acg gtc atg ggc ggg tcg agt ggc 1296
Leu Leu Ile Gly Glu Arg Leu Ala Thr Val Met Gly Gly Ser Ser Gly
420 425 430
gtc ctg atg tcg atc ttc ttc aca gct gcc gga cag aaa atg cat gac 1344
Val Leu Met Ser Ile Phe Phe Thr Ala Ala Gly Gln Lys Met His Asp
435 440 445
gga aaa tca ctg ccg gag gca ttg ctg agt ggg ctt gcg caa atg aag 1392
Gly Lys Ser Leu Pro Glu Ala Leu Leu Ser Gly Leu Ala Gln Met Lys
450 455 460
cat tac ggc gga gcg gat ctt ggc gat cgt acc ttg atc gac gcg cta 1440
His Tyr Gly Gly Ala Asp Leu Gly Asp Arg Thr Leu Ile Asp Ala Leu
465 470 475 480
cag cct gca ctg gag acg ctg cat aac ggc gat att cag gcg gct gcc 1488
Gln Pro Ala Leu Glu Thr Leu His Asn Gly Asp Ile Gln Ala Ala Ala
485 490 495
cag gca gcg aaa aaa ggc gca gac gct acg gct ggc atg caa aaa gcg 1536
Gln Ala Ala Lys Lys Gly Ala Asp Ala Thr Ala Gly Met Gln Lys Ala
500 505 510
gga gca ggg cgt tcg tcg tat gtg aat aaa gag aac ctg gaa ggt gta 1584
Gly Ala Gly Arg Ser Ser Tyr Val Asn Lys Glu Asn Leu Glu Gly Val
515 520 525
ata gat cct ggg gca gtg gcc gtt gca gag gtg ttt gcg gca gtg gcc 1632
Ile Asp Pro Gly Ala Val Ala Val Ala Glu Val Phe Ala Ala Val Ala
530 535 540
aaa gca aaa cag tag 1647
Lys Ala Lys Gln
545
<210>51
<211>548
<212>PRT
<213>肠杆菌属菌种638
<400>51
Met Ser Arg Phe Phe Phe Asn Asp Arg Lys Gln Leu Val Asn Asp Ala
1 5 10 15
Ile Glu Gly Ile Leu Ile Ser Ala Pro His Gly Asn Leu Val Lys Leu
20 25 30
Asp Ile Asp Pro Ala Ile Arg Val Val Ala Arg Ser Asp Trp Asp Lys
35 40 45
Ser Arg Val Ala Val Ile Ser Gly Gly Gly Ser Gly His Glu Pro Ala
50 55 60
His Ala Gly Phe Val Gly Lys Gly Met Leu Thr Ala Ala Val Cys Gly
65 70 75 80
Asp Leu Phe Ala Ser Pro Ser Val Asp Ala Val Leu Asn Ala Ile Val
85 90 95
Ala Val Thr Gly Asp Arg Gly Cys Leu Leu Ile Val Lys Asn Tyr Thr
100 105 110
Gly Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Lys Ala Lys Arg Tyr
115 120 125
Gly Leu Lys Val Glu Met Val Ile Val Ala Asp Asp Ile Ala Leu Pro
130 135 140
Asp Asn Lys Gln Pro Arg Gly Ile Ala Gly Thr Ala Leu Val His Lys
145 150 155 160
Ile Ala Gly Tyr Ala Ala Glu Gln Gly Lys Ser Leu Ala Asp Val Arg
165 170 175
Asp Ile Ala Gln Gln Ala Cys Asp Asn Ile Trp Ser Leu Gly Val Ala
180 185 190
Met Gln Thr Cys Asn Leu Pro Gly Ser Asp Asp Glu Glu Gly Arg Ile
195 200 205
Lys Asp Gly His Val Glu Leu Gly Leu Gly Ile His Gly Glu Pro Gly
210 215 220
Ala Ser Val Val Asp Thr His Asn Ser Lys Glu Ile Ile Asp Thr Leu
225 230 235 240
Val Lys Pro Leu Lys Glu Thr Ala Gly Glu Gly Lys Phe Ala Val Leu
245 250 255
Ile Asn Asn Leu Gly Gly Val Ser Ala Leu Glu Met Ala Leu Leu Thr
260 265 270
Lys Glu Leu Ala Asp Ser Ala Leu Lys Glu Asn Ile Ala Tyr Leu Ile
275 280 285
Gly Pro Ala Pro Leu Val Ser Ser Leu Asp Met Lys Gly Phe Ser Leu
290 295 300
Ser Leu Leu Gln Leu Asn Asp Thr Phe Glu Lys Ala Ile Asn Ala Pro
305 310 315 320
Val Glu Thr Ile Gly Trp Gln Lys Pro Val Ala Phe Ala Pro Leu Arg
325 330 335
Thr Leu Ser His Thr Ala Ile Gln Asp Arg Val Glu Phe Thr Pro Ser
340 345 350
Gly Asn Asp Glu Val Ala Ala Arg Val Ala Ala Ala Thr Gln Thr Leu
355 360 365
Leu Ala Leu Glu Asn Arg Leu Asn Ala Leu Asp Ala Lys Val Gly Asp
370 375 380
Gly Asp Thr Gly Ser Thr Phe Ala Gln Gly Ala Arg Glu Ile Ala Gln
385 390 395 400
Leu Leu Glu Gln Lys Gln Leu Pro Leu Asn Asp Leu Ser Lys Leu Leu
405 410 415
Leu Leu Ile Gly Glu Arg Leu Ala Thr Val Met Gly Gly Ser Ser Gly
420 425 430
Val Leu Met Ser Ile Phe Phe Thr Ala Ala Gly Gln Lys Met His Asp
435 440 445
Gly Lys Ser Leu Pro Glu Ala Leu Leu Ser Gly Leu Ala Gln Met Lys
450 455 460
His Tyr Gly Gly Ala Asp Leu Gly Asp Arg Thr Leu Ile Asp Ala Leu
465 470 475 480
Gln Pro Ala Leu Glu Thr Leu His Asn Gly Asp Ile Gln Ala Ala Ala
485 490 495
Gln Ala Ala Lys Lys Gly Ala Asp Ala Thr Ala Gly Met Gln Lys Ala
500 505 510
Gly Ala Gly Arg Ser Ser Tyr Val Asn Lys Glu Asn Leu Glu Gly Val
515 520 525
Ile AspPro Gly Ala Val Ala Val Ala Glu Val Phe Ala Ala Val Ala
530 535 540
Lys Ala Lys Gln
545
<210>52
<211>1695
<212>DNA
<213>假单胞菌属菌种CNPT3
<220>
<221>CDS
<222>(1)..(1695)
<223>
<400>52
atg gtt ata tta ttt ttt aac cta ctt tat ttt aaa cta ctt ttt gga 48
Met Val Ile Leu Phe Phe Asn Leu Leu Tyr Phe Lys Leu Leu Phe Gly
1 5 10 15
gta att atg agc cgt tta ttt att aat gat aaa gca acg tta gtc cat 96
Val Ile Met Ser Arg Leu Phe Ile Asn Asp Lys Ala Thr Leu Val His
20 25 30
gat gcg att gat ggc atc tta tat agc aat aag cat aat aac ctt gtt 144
Asp Ala Ile Asp Gly Ile Leu Tyr Ser Asn Lys His Asn Asn Leu Val
35 40 45
cgt ttg gat gtt gat ccg caa att aga att gtg acg cgt aat gat tgg 192
Arg Leu Asp Val Asp Pro Gln Ile Arg Ile Val Thr Arg Asn Asp Trp
50 55 60
cat cac gat aaa gtt gcc att atc tct ggt ggt ggc tct ggg cat gag 240
His His Asp Lys Val Ala Ile Ile Ser Gly Gly Gly Ser Gly His Glu
65 70 75 80
cct gct cat gtt ggt ttt att ggt aaa ggt atg tta acg gct gct gtt 288
Pro Ala His Val Gly Phe Ile Gly Lys Gly Met Leu Thr Ala Ala Val
85 90 95
tgt ggc gat gtt ttt gcc tca cca agt gtg gat gcg gtg tta aat gcc 336
Cys Gly Asp Val Phe Ala Ser Pro Ser Val Asp Ala Val Leu Asn Ala
100 105 110
ata gtc cat gtc aca gga gag aaa ggt tgt ttg gtc att gtt aaa aac 384
Ile Val His Val Thr Gly Glu Lys Gly Cys Leu Val Ile Val Lys Asn
115 120 125
tat acc ggc gac cgt ttg aac ttt ggc ctt gct tgt gaa aaa gcc aaa 432
Tyr Thr Gly Asp Arg Leu Asn Phe Gly Leu Ala Cys Glu Lys Ala Lys
130 135 140
aaa atg ggt tta aac gtt gaa atg gta ata gta gac gat gat atc tct 480
Lys Met Gly Leu Asn Val Glu Met Val Ile Val Asp Asp Asp Ile Ser
145 150 155 160
att cct gac aat ctt aaa cct cgt ggt att gct ggc aca ttg ttt gtc 528
Ile Pro Asp Asn Leu Lys Pro Arg Gly Ile Ala Gly Thr Leu Phe Val
165 170 175
cat aaa gtg gca ggt aat gca gca gag caa ggc gct tct tta aat gtc 576
His Lys Val Ala Gly Asn Ala Ala Glu Gln Gly Ala Ser Leu Asn Val
180 185 190
gtt aaa aag gca gcc caa ggt gcg att gat gca acg gca agt att ggc 624
Val Lys Lys Ala Ala Gln Gly Ala Ile Asp Ala Thr Ala Ser Ile Gly
195 200 205
ctc gca ctg aca agt tgc tct tta ccg gga gaa gaa tca acg cag cgt 672
Leu Ala Leu Thr Ser Cys Ser Leu Pro Gly Glu Glu Ser Thr Gln Arg
210 215 220
att gca gaa ggc aag gct gaa tta ggg tta ggt att cat gga gag cct 720
Ile Ala Glu Gly Lys Ala Glu Leu Gly Leu Gly Ile His Gly Glu Pro
225 230 235 240
ggc att aaa aca att gat gta aca tgt tgt cgt gat ctt gtg atg atc 768
Gly Ile Lys Thr Ile Asp Val Thr Cys Cys Arg Asp Leu Val Met Ile
245 250 255
atg gtt gat aaa cta aaa caa tcg ttt tct gct cca gat att aaa att 816
Met Val Asp Lys Leu Lys Gln Ser Phe Ser Ala Pro AspIle Lys Ile
260 265 270
gca gtg atg atc aac aat tta ggt ggc gtt tct cca tta gag atg agc 864
Ala Val Met Ile Asn Asn Leu Gly Gly Val Ser Pro Leu Glu Met Ser
275 280 285
ctt ata tgt aaa gac atc gtg gaa tct gag tta aaa aat aat att gag 912
Leu Ile Cys Lys Asp Ile Val Glu Ser Glu Leu Lys Asn Asn Ile Glu
290 295 300
tta gtg gtt ggg cct gct cca ttt atg acg gct att gat atg aaa gga 960
Leu Val Val Gly Pro Ala Pro Phe Met Thr Ala Ile Asp Met Lys Gly
305 310 315 320
ttt tca att tca gtg att gaa tta aca ggc gat cat gct caa gct ctg 1008
Phe Ser Ile Ser Val Ile Glu Leu Thr Gly Asp His Ala Gln Ala Leu
325 330 335
tgc gcc cct gtt gaa gtg gat gca tgg gtt gaa gcc att cca ttg cgc 1056
Cys Ala Pro Val Glu Val Asp Ala Trp Val Glu Ala Ile Pro Leu Arg
340 345 350
cca tta aat gtg ata aaa aaa gat aaa gtg tcc att aac ttt gca ttt 1104
Pro Leu Asn Val Ile Lys Lys Asp Lys Val Ser Ile Asn Phe Ala Phe
355 360 365
gaa gcg tct gaa aat gca cag gtt gca agt att gtt aaa aca gta acc 1152
Glu Ala Ser Glu Asn Ala Gln Val Ala Ser Ile Val Lys Thr Val Thr
370 375 380
ttg gct ttg att aat gca gaa aaa gag tta aat cgt ctg gat acg tta 1200
Leu Ala Leu Ile Asn Ala Glu Lys Glu Leu Asn Arg Leu Asp Thr Leu
385 390 395 400
gtg ggc gat ggg gat acc ggt tca acg ttc tct gcg ggt gct cga caa 1248
Val Gly Asp Gly Asp Thr Gly Ser Thr Phe Ser Ala Gly Ala Arg Gln
405 410 415
gtt tta gct gag ctc aat gcg ggt aat tta cca ctt aat gat act ggc 1296
Val Leu Ala Glu Leu Asn Ala Gly Asn Leu Pro Leu Asn Asp Thr Gly
420 425 430
gcg ttg ctt aat gtc att ggg gaa caa ctt gct acc gtt atg ggg gga 1344
Ala Leu Leu Asn Val Ile Gly Glu Gln Leu Ala Thr Val Met Gly Gly
435 440 445
tcg tca ggc gta tta ttc tct atc ttc ttc aca gca gcg ggt cat cat 1392
Ser Ser Gly Val Leu Phe Ser Ile Phe Phe Thr Ala Ala Gly His His
450 455 460
tac cag caa cat ggt gat aca gta caa gca tta caa gcc ggt tta caa 1440
Tyr Gln Gln His Gly Asp Thr Val Gln Ala Leu Gln Ala Gly Leu Gln
465 470 475 480
caa atg atg caa tac ggt gga gca aaa ccg ggt gat cgt acg atg att 1488
Gln Met Met Gln Tyr Gly Gly Ala Lys Pro Gly Asp Arg Thr Met Ile
485 490 495
gat gca atg tac cca gcc ttt atc gct tgg aaa aat gaa ggt ttt gaa 1536
Asp Ala Met Tyr Pro Ala Phe Ile Ala Trp Lys Asn Glu Gly Phe Glu
500 505 510
gct gcc att gtt gcg gct aaa ata ggg gca gaa agc aca gcc act atg 1584
Ala Ala Ile Val Ala Ala Lys Ile Gly Ala Glu Ser Thr Ala Thr Met
515 520 525
gtc gaa gcc aaa gca gga cgc tct tct tat tta aac agt gag agt tta 1632
Val Glu Ala Lys Ala Gly Arg Ser Ser Tyr Leu Asn Ser Glu Ser Leu
530 535 540
aaa ggt gtt aaa gat ccg ggc tca gtc gct gtt gag ttg gtt ttt gac 1680
Lys Gly Val Lys Asp Pro Gly Ser Val Ala Val Glu Leu Val Phe Asp
545 550 555 560
gcg ttt aat gtg taa 1695
Ala Phe Asn Val
<210>53
<211>564
<212>PRT
<213>假单胞菌属菌种CNPT3
<400>53
Met Val Ile Leu Phe Phe Asn Leu Leu Tyr Phe Lys Leu Leu Phe Gly
1 5 10 15
Val Ile Met Ser Arg Leu Phe Ile Asn Asp Lys Ala Thr Leu Val His
20 25 30
Asp Ala Ile Asp Gly Ile Leu Tyr Ser Asn Lys His Asn Asn Leu Val
35 40 45
Arg Leu Asp Val Asp Pro Gln Ile Arg Ile Val Thr Arg Asn Asp Trp
50 55 60
His His Asp Lys Val Ala Ile Ile Ser Gly Gly Gly Ser Gly His Glu
65 70 75 80
Pro Ala His Val Gly Phe Ile Gly Lys Gly Met Leu Thr Ala Ala Val
85 90 95
Cys Gly Asp Val Phe Ala Ser Pro Ser Val Asp Ala Val Leu Asn Ala
100 105 110
Ile Val His Val Thr Gly Glu Lys Gly Cys Leu Val Ile Val Lys Asn
115 120 125
Tyr Thr Gly Asp Arg Leu Asn Phe Gly Leu Ala Cys Glu Lys Ala Lys
130 135 140
Lys Met Gly Leu Asn Val Glu Met Val Ile Val Asp Asp Asp Ile Ser
145 150 155 160
Ile Pro Asp Asn Leu Lys Pro Arg Gly Ile Ala Gly Thr Leu Phe Val
165 170 175
His Lys Val Ala Gly Asn Ala Ala Glu Gln Gly Ala Ser Leu Asn Val
180 185 190
Val Lys Lys Ala Ala Gln Gly Ala Ile Asp Ala Thr Ala Ser Ile Gly
195 200 205
Leu Ala Leu Thr Ser Cys Ser Leu Pro Gly Glu Glu Ser Thr Gln Arg
210 215 220
Ile Ala Glu Gly Lys Ala Glu Leu Gly Leu Gly Ile His Gly Glu Pro
225 230 235 240
Gly Ile Lys Thr Ile Asp Val Thr Cys Cys Arg Asp Leu Val Met Ile
245 250 255
Met Val Asp Lys Leu Lys Gln Ser Phe Ser Ala Pro Asp Ile Lys Ile
260 265 270
Ala Val Met Ile Asn Asn Leu Gly Gly Val Ser Pro Leu Glu Met Ser
275 280 285
Leu Ile Cys Lys Asp Ile Val Glu Ser Glu Leu Lys Asn Asn Ile Glu
290 295 300
Leu Val Val Gly Pro Ala Pro Phe Met Thr Ala Ile Asp Met Lys Gly
305 310 315 320
Phe Ser Ile Ser Val Ile Glu Leu Thr Gly Asp His Ala Gln Ala Leu
325 330 335
Cys Ala Pro Val Glu Val Asp Ala Trp Val Glu Ala Ile Pro Leu Arg
340 345 350
Pro Leu Asn Val Ile Lys Lys Asp Lys Val Ser Ile Asn Phe Ala Phe
355 360 365
Glu Ala Ser Glu Asn Ala Gln Val Ala Ser Ile Val Lys Thr Val Thr
370 375 380
Leu Ala Leu Ile Asn Ala Glu Lys Glu Leu Asn Arg Leu Asp Thr Leu
385 390 395 400
Val Gly Asp Gly Asp Thr Gly Ser Thr Phe Ser Ala Gly Ala Arg Gln
405 410 415
Val Leu Ala Glu Leu Asn Ala Gly Asn Leu Pro Leu Asn Asp Thr Gly
420 425 430
Ala Leu Leu Asn Val Ile Gly Glu Gln Leu Ala Thr Val Met Gly Gly
435 440 445
Ser Ser Gly Val Leu Phe Ser Ile Phe Phe Thr Ala Ala Gly His His
450 455 460
Tyr Gln Gln His Gly Asp Thr Val Gln Ala Leu Gln Ala Gly Leu Gln
465 470 475 480
Gln Met Met Gln Tyr Gly Gly Ala Lys Pro Gly Asp Arg Thr Met Ile
485 490 495
Asp Ala Met Tyr Pro Ala Phe Ile Ala Trp Lys Asn Glu Gly Phe Glu
500 505 510
Ala Ala Ile Val Ala Ala Lys Ile Gly Ala Glu Ser Thr Ala Thr Met
515 520 525
Val Glu Ala Lys Ala Gly Arg Ser Ser Tyr Leu Asn Ser Glu Ser Leu
530 535 540
Lys Gly Val Lys Asp Pro Gly Ser Val Ala Val Glu Leu Val Phe Asp
545 550 555 560
Ala Phe Asn Val
<210>54
<211>1647
<212>DNA
<213>Stapia aggregata IAMl2614
<220>
<221>CDSl
<222>(1)..(1647)
<223>
<220>
<22l>CDS
<222>(1)..(1647)
<223>
<400>54
atg aag caa ttc atc aat acc aag gaa acg ctc gtc acc gaa gcg att 48
Met Lys Gln Phe Ile Asn Thr Lys Glu Thr Leu Val Thr Glu Ala Ile
1 5 10 15
gac ggc atg ttg cgc acg gcc ggc ggg cgg ctt gcc cgg ctt gac ggc 96
Asp Gly Met Leu Arg Thr Ala Gly Gly Arg Leu Ala Arg Leu Asp Gly
20 25 30
tat ccg cat atc aag gtg gtc gtg cgc acc gac tgg gac aaa tcg aag 144
Tyr Pro His Ile Lys Val Val Val Arg Thr Asp Trp Asp Lys Ser Lys
35 40 45
gtg gct ctg gtg tcc ggc ggc ggt tcc ggc cac gag ccg agc cat gcc 192
Val Ala Leu Val Ser Gly Gly Gly Ser Gly His Glu Pro Ser His Ala
50 55 60
ggt ttc gtc ggc cag ggc atg ctg acg gcg gcc gtc tgc ggc gaa gtg 240
Gly Phe Val Gly Gln Gly Met Leu Thr Ala Ala Val Cys Gly Glu Val
65 70 75 80
ttt gcc tcg cct tcc gtg gat gcg gtg ctg gcg ggc att ctg gcc gtc 288
Phe Ala Ser Pro Ser Val Asp Ala Val Leu Ala Gly Ile Leu Ala Val
85 90 95
acc ggc aag gct ggc tgc ctg ctg atc gtc aag aac tac acc ggc gac 336
Thr Gly Lys Ala Gly Cys Leu Leu Ile Val Lys Asn Tyr Thr Gly Asp
100 105 110
cgg ctg aac ttc ggt ctg gcc gcc gag cgg gcc cgc tcc ttc gga ctg 384
Arg Leu Asn Phe Gly Leu Ala Ala Glu Arg Ala Arg Ser Phe Gly Leu
115 120 125
aag gtc aac atg gtg atc gtc gac gac gac gtt gcc ctg ccg gac ctg 432
Lys Val Asn Met Val Ile Val Asp Asp Asp Val Ala Leu Pro Asp Leu
130 135 140
ccg cag gcg cgc ggt gtc gcc ggc acg ctg ttc gtg cac aag atc gcc 480
Pro Gln Ala Arg Gly Val Ala Gly Thr Leu Phe Val His Lys Ile Ala
145 150 155 160
gga gcg ctt gcc gat cag ggt gcg gat ctg gaa acc atc acg gag gcc 528
Gly Ala Leu Ala Asp Gln Gly Ala Asp Leu Glu Thr Ile Thr Glu Ala
165 170 175
gcc agg aaa acc atc ggc ggt gcg att tcc atc ggc atg tcg ctg gac 576
Ala Arg Lys Thr Ile Gly Gly Ala Ile Ser Ile Gly Met Ser Leu Asp
180 185 190
acc tgc acg gtg ccg gga tcg ccc aag gaa gac cgc att gca cac ggc 624
Thr Cys Thr Val Pro Gly Ser Pro Lys Glu Asp Arg Ile Ala His Gly
195 200 205
aag gcg gaa ctc gga ctt ggc att cac ggc gag gcg ggg atc gag cag 672
Lys Ala Glu Leu Gly Leu Gly Ile His Gly Glu Ala Gly Ile Glu Gln
210 215 220
gtc gac tat tcc aac gcc cgc gcg gcc atg gcc atg gtg gtg gac cgg 720
Val Asp Tyr Ser Asn Ala Arg Ala Ala Met Ala Met Val Val Asp Arg
225 230 235 240
ctg gcg ccg aac ctc tcg ccc gga ccg cat gtg gcg atc ctc aac aat 768
Leu Ala Pro Asn Leu Ser Pro Gly Pro His Val Ala Ile Leu Asn Asn
245 250 255
ctg ggc agc acg acg ccg ctg gaa atg tcg gtg ctt ctg gaa gaa ctc 816
Leu Gly Ser Thr Thr Pro Leu Glu Met Ser Val Leu Leu Glu Glu Leu
260 265 270
acg gct tcg cgc atc ggc agc cag atc cgc tgg gtc atc ggc ccg gcg 864
Thr Ala Ser Arg Ile Gly Ser Gln Ile Arg Trp Val Ile Gly Pro Ala
275 280 285
gcg atg atg acc tcg ctc gac atg cat ggg ttc tcc gtg tcg ctg ctg 912
Ala Met Met Thr Ser Leu Asp Met His Gly Phe Ser Val Ser Leu Leu
290 295 300
ccg gtc ggc aag acc gaa gaa gcc ttg ctg cag gcc ccg gtc gcg ccc 960
Pro Val Gly Lys Thr Glu Glu Ala Leu Leu Gln Ala Pro Val Ala Pro
305 310 315 320
tgg gca tgg ccc ggc tgc ctt gcg ctt ggc gca gtg tcc gtg ctg ccg 1008
Trp Ala Trp Pro Gly Cys Leu Ala Leu Gly Ala Val Ser Val Leu Pro
325 330 335
cta ccg gac ggc ctg acg ccg atc cag ccg ctg ccg tcc aag aac ccg 1056
Leu Pro Asp Gly Leu Thr Pro Ile Gln Pro Leu Pro Ser Lys Asn Pro
340 345 350
gag acg cgg aag ttc atc gag cgc tgc tgc gat atc ctg atc gcc gcc 1104
Glu Thr Arg Lys Phe Ile Glu Arg Cys Cys Asp Ile Leu Ile Ala Ala
355 360 365
gag gac gac ctc aat gcg ctc gac gcc aag tcg ggc gac ggc gac acc 1152
Glu Asp Asp Leu Asn Ala Leu Asp Ala Lys Ser Gly Asp Gly Asp Thr
370 375 380
ggc agc acg ctt gcc acc gcc gcc cgg gcg ctg gtg aag gcg ctc gac 1200
Gly Ser Thr Leu Ala Thr Ala Ala Arg Ala Leu Val Lys Ala Leu Asp
385 390 395 400
cgg ctg ccg ctg gcg gat ctc acc cag ctc tac cgg gcc atc ggg ctg 1248
Arg Leu Pro Leu Ala Asp Leu Thr Gln Leu Tyr Arg Ala Ile Gly Leu
405 410 415
gaa ctc agc cag acc atg ggc ggg tct tcc ggg gtg ctg ctg gcg atc 1296
Glu Leu Ser Gln Thr Met Gly Gly Ser Ser Gly Val Leu Leu Ala Ile
420 425 430
ttc ttt gcc gct gcg ggc gat gcg tcg tcc agc ggg cgc ggg gcc atc 1344
Phe Phe Ala Ala Ala Gly Asp Ala Ser Ser Ser Gly Arg Gly Ala Ile
435 440 445
gga gcc ctg aag gcc ggt ctt gac cgg atc atg cag gtg ggc ggc gcc 1392
Gly Ala Leu Lys Ala Gly Leu Asp Arg Ile Met Gln Val Gly Gly Ala
450 455 460
cag ccg ggc gac cgc acc atg atc gac gcg ctg ctg ccg gca ctg aat 1440
Gln Pro Gly Asp Arg Thr Met Ile Asp Ala Leu Leu Pro Ala Leu Asn
465 470 475 480
gcg ctg gaa aac ggt atc gag gct gcg gcg agc gag gcc cgt cag ggg 1488
Ala Leu Glu Asn Gly Ile Glu Ala Ala Ala Ser Glu Ala Arg Gln Gly
485 490 495
gcg gat gcg acg tcg cgg atc acg cgg gca cgc gcg ggc agg gcg tct 1536
Ala Asp Ala Thr Ser Arg Ile Thr Arg Ala Arg Ala Gly Arg Ala Ser
500 505 510
tat gtc tcc gag gcc agc ctc tcc gga cac aac gat ccg ggc gcg gaa 1584
Tyr Val Ser Glu Ala Ser Leu Ser Gly His Asn Asp Pro Gly Ala Glu
515 520 525
gcc gtt gcg cgg ctg ttc gag caa ttg acc ctt tct ccg gcc ctt acc 1632
Ala Val Ala Arg Leu Phe Glu Gln Leu Thr Leu Ser Pro Ala Leu Thr
530 535 540
tcc aag tcc gca tag 1647
Ser Lys Ser Ala
545
<210>55
<211>548
<212>PRT
<213>Stapia aggregata IAMl2614
<400>55
Met Lys Gln Phe Ile Asn Thr Lys Glu Thr Leu Val Thr Glu Ala Ile
1 5 10 15
Asp Gly Met Leu Arg Thr Ala Gly Gly Arg Leu Ala Arg Leu Asp Gly
20 25 30
Tyr Pro His Ile Lys Val Val Val Arg Thr Asp Trp Asp Lys Ser Lys
35 40 45
Val Ala Leu Val Ser Gly Gly Gly Ser Gly His Glu Pro Ser His Ala
50 55 60
Gly Phe Val Gly Gln Gly Met Leu Thr Ala Ala Val Cys Gly Glu Val
65 70 75 80
Phe Ala Ser Pro Ser Val Asp Ala Val Leu Ala Gly Ile Leu Ala Val
85 90 95
Thr Gly Lys Ala Gly Cys Leu Leu Ile Val Lys Asn Tyr Thr Gly Asp
100 105 110
Arg Leu Asn Phe Gly Leu Ala Ala Glu Arg Ala Arg Ser Phe Gly Leu
115 120 125
Lys Val Asn Met Val Ile Val Asp Asp Asp Val Ala Leu Pro Asp Leu
130 135 140
Pro Gln Ala Arg Gly Val Ala Gly Thr Leu Phe Val His Lys Ile Ala
145 150 155 160
Gly Ala Leu Ala Asp Gln Gly Ala Asp Leu Glu Thr Ile Thr Glu Ala
165 170 175
Ala Arg Lys Thr Ile Gly Gly Ala Ile Ser Ile Gly Met Ser Leu Asp
180 185 190
Thr Cys Thr Val Pro Gly Ser Pro Lys Glu Asp Arg Ile Ala His Gly
195 200 205
Lys Ala Glu Leu Gly Leu Gly Ile His Gly Glu Ala Gly Ile Glu Gln
210 215 220
Val Asp Tyr Ser Asn Ala Arg Ala Ala Met Ala Met Val Val Asp Arg
225 230 235 240
Leu Ala Pro Asn Leu Ser Pro Gly Pro His Val Ala Ile Leu Asn Asn
245 250 255
Leu Gly Ser Thr Thr Pro Leu Glu Met Ser Val Leu Leu Glu Glu Leu
260 265 270
Thr Ala Ser Arg Ile Gly Ser Gln Ile Arg Trp Val Ile Gly Pro Ala
275 280 285
Ala Met Met Thr Ser Leu Asp Met His Gly Phe Ser Val Ser Leu Leu
290 295 300
Pro Val Gly Lys Thr Glu Glu Ala Leu Leu Gln Ala Pro Val Ala Pro
305 310 315 320
Trp Ala Trp Pro Gly Cys Leu Ala Leu Gly Ala Val Ser Val Leu Pro
325 330 335
Leu Pro Asp Gly Leu Thr Pro Ile Gln Pro Leu Pro Ser Lys Asn Pro
340 345 350
Glu Thr Arg Lys Phe Ile Glu Arg Cys Cys Asp Ile Leu Ile Ala Ala
355 360 365
Glu Asp Asp Leu Asn Ala Leu Asp Ala Lys Ser Gly Asp Gly Asp Thr
370 375 380
Gly Ser Thr Leu Ala Thr Ala Ala Arg Ala Leu Val Lys Ala Leu Asp
385 390 395 400
Arg Leu Pro Leu Ala Asp Leu Thr Gln Leu Tyr Arg Ala Ile Gly Leu
405 410 415
Glu Leu Ser Gln Thr Met Gly Gly Ser Ser Gly Val Leu Leu Ala Ile
420 425 430
Phe Phe Ala Ala Ala Gly Asp Ala Ser Ser Ser Gly Arg Gly Ala Ile
435 440 445
Gly Ala Leu Lys Ala Gly Leu Asp Arg Ile Met Gln Val Gly Gly Ala
450 455 460
Gln Pro Gly Asp Arg Thr Met Ile Asp Ala Leu Leu Pro Ala Leu Asn
465 470 475 480
Ala Leu Glu Asn Gly Ile Glu Ala Ala Ala Ser Glu Ala Arg Gln Gly
485 490 495
Ala Asp Ala Thr Ser Arg Ile Thr Arg Ala Arg Ala Gly Arg Ala Ser
500 505 510
Tyr Val Ser Glu Ala Ser Leu Ser Gly His Asn Asp Pro Gly Ala Glu
515 520 525
Ala Val Ala Arg Leu Phe Glu Gln Leu Thr Leu Ser Pro Ala Leu Thr
530 535 540
Ser Lys Ser Ala
545
<210>56
<211>1641
<212>DNA
<213>豌豆根瘤菌(Rhizobium leguminosarum)
<220>
<221>CDS
<222>(1)..(1641)
<223>
<400>56
atg aaa cac ttc ttc aac cgc agg gaa aac atc gtc acc gaa gcc ttg 48
Met Lys His Phe Phe Asn Arg Arg Glu Asn Ile Val Thr Glu Ala Leu
1 5 10 15
gac ggt ctg ctt ctg acg agc agc aag ggt cgt ctt gcc cgc ctc gac 96
Asp Gly Leu Leu Leu Thr Ser Ser Lys Gly Arg Leu Ala Arg Leu Asp
20 25 30
agc ttt ccc gac atc aag gtg atc ctg cgc gct gac tgg gac aag tcg 144
Ser Phe Pro Asp Ile Lys Val Ile Leu Arg Ala Asp Trp Asp Lys Ser
35 40 45
aag gtg gcg atc atc tca ggc ggc ggc gcc ggt cat gag ccc tcc cat 192
Lys Val Ala Ile Ile Ser Gly Gly Gly Ala Gly His Glu Pro Ser His
50 55 60
gcc ggc ttc gtc ggt aag ggc atg ctg acg gct gcc gta tcc ggc gag 240
Ala Gly Phe Val Gly Lys Gly Met Leu Thr Ala Ala Val Ser Gly Glu
65 70 75 80
att ttc gcc tcg ccg agc gtc gat gcc gtg ctg aca gcg atc cgc gcc 288
Ile Phe Ala Ser Pro Ser Val Asp Ala Val Leu Thr Ala Ile Arg Ala
85 90 95
gtc gcc ggc gaa aag ggc gcc ttg ctg atc gtc aag aac tat acc ggc 336
Val Ala Gly Glu Lys Gly Ala Leu Leu Ile Val Lys Asn Tyr Thr Gly
100 105 110
gac cgg ctg aat ttc ggc ctc gcc gcc gag aag gcg cgc gcc gaa ggt 384
Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Lys Ala Arg Ala Glu Gly
115 120 125
ttc gac gtc gaa atg gtc atc gtc gcc gac gat atc gcc atc ccc gag 432
Phe Asp Val Glu Met Val Ile Val Ala Asp Asp Ile Ala Ile Pro Glu
130 135 140
atc aac cag ccg cgc ggc gtc gcc ggg act ctg ttc gtc cac aag atc 480
Ile Asn Gln Pro Arg Gly Val Ala Gly Thr Leu Phe Val His Lys Ile
145 150 155 160
gct ggc tat cac gcc gaa agg ggc gag gac ctg aag acg gtc gca gcc 528
Ala Gly Tyr His Ala Glu Arg Gly Glu Asp Leu Lys Thr Val Ala Ala
165 170 175
cat gcc gcg gca gcg gcc ggc gac atc gtc tcg ctc ggc atg tct ctg 576
His Ala Ala Ala Ala Ala Gly Asp Ile Val Ser Leu Gly Met Ser Leu
180 185 190
tcc acc tgc agc gtg ccc ggc cag gcg cat gag agc cgc ctc ggc gag 624
Ser Thr Cys Ser Val Pro Gly Gln Ala His Glu Ser Arg Leu Gly Glu
195 200 205
aac gag ggc gaa ctc ggt ctc ggc atc cat ggc gag ccc ggc gtc gag 672
Asn Glu Gly Glu Leu Gly Leu Gly Ile His Gly Glu Pro Gly Val Glu
210 215 220
cgc att gcg ctg cag ccg gtc gtc gat atc gtc gcc acc atg gtg gcg 720
Arg Ile Ala Leu Gln Pro Val Val Asp Ile Val Ala Thr Met Val Ala
225 230 235 240
cgc cta tcg cct gcg ctg cgc gaa ggg gga aac cac gcc ctt ctc atc 768
Arg Leu Ser Pro Ala Leu Arg Glu Gly Gly Asn His Ala Leu Leu Ile
245 250 255
aac aat ctc ggc gcc gta ccg ccg ctc gaa atg acc gtt att gcc aat 816
Asn Asn Leu Gly Ala Val Pro Pro Leu Glu Met Thr Val Ile Ala Asn
260 265 270
gtg gtg ctg tcc tcg tcg ctt gcc gat cgc gtc agg ctg atc atc ggc 864
Val Val Leu Ser Ser Ser Leu Ala Asp Arg Val Arg Leu Ile Ile Gly
275 280 285
ccg gcg ccg atg atg acc gcg ctc aac atg aac ggc ttc tcg ctg tcg 912
Pro Ala Pro Met Met Thr Ala Leu Asn Met Asn Gly Phe Ser Leu Ser
290 295 300
ctg atc cga ctg gat gcc gct cgc gag gcg gcg ctg acg gca gcg gtc 960
Leu Ile Arg Leu Asp Ala Ala Arg Glu Ala Ala Leu Thr Ala Ala Val
305 310 315 320
gaa ccg cat gcc cgg atg cca gcc gtc gaa cgc cac gag atc agg gtc 1008
Glu Pro His Ala Trp Met Pro Ala Val Glu Arg His Glu Ile Arg Val
325 330 335
atc gcc gca ccg cga aca tca gcc gga ctg aac ggc gcg cca gtg gcc 1056
Ile Ala Ala Pro Arg Thr Ser Ala Gly Leu Asn Gly Ala Pro Val Ala
340 345 350
ggg gat aat ctc cgc aac cgg cgt ctg atc aca gcg ctc tgc gag cat 1104
Gly Asp Asn Leu Arg Asn Arg Arg Leu Ile Thr Ala Leu Cys Glu His
355 360 365
ctg atc tcg cag gaa agc gaa ctc aac cgg ctg gat ggc cgc gtc ggc 1152
Leu Ile Ser Gln Glu Ser Glu Leu Asn Arg Leu Asp Gly Arg Val Gly
370 375 380
gac ggt gat acc ggc tcg acg gtg gcg aca ggc gcc cgc agc gtg ctt 1200
Asp Gly Asp Thr Gly Ser Thr Val Ala Thr Gly Ala Arg Ser Val Leu
385 390 395 400
gcc cgc ctg gac acg ctg ccg ctt gat cgg ccg gct gca acg ctt gcc 1248
Ala Arg Leu Asp Thr Leu Pro Leu Asp Arg Pro Ala Ala Thr Leu Ala
405 410 415
tcg ctc ggc gac atc ctc ggc acc agc atg ggc gga tcg agc ggc gtg 1296
Ser Leu Gly Asp Ile Leu Gly Thr Ser Met Gly Gly Ser Ser Gly Val
420 425 430
ctg ctg tcg atc ttc ttc acc gca gcg gca aag gcg atg gcc gac aag 1344
Leu Leu Ser Ile Phe Phe Thr Ala Ala Ala Lys Ala Met Ala Asp Lys
435 440 445
gcc gat ata tca gca gcc ctt att gcc ggg ctc gac agg atg acg ttc 1392
Ala Asp Ile Ser Ala Ala Leu Ile Ala Gly Leu Asp Arg Met Thr Phe
450 455 460
tat ggc gga gcc gaa gtc ggc gac cgg acg atg gtc gat gcg ctg tcg 1440
Tyr Gly Gly Ala Glu Val Gly Asp Arg Thr Met Val Asp Ala Leu Ser
465 470 475 480
cct gcc ctg cag gcg ctc gca tcc ggc gat gtc gcg gca gcg gcc agg 1488
Pro Ala Leu Gln Ala Leu Ala Ser Gly Asp Val Ala Ala Ala Ala Arg
485 490 495
gct gct gcc gca ggt gcg gag tcg acg aag acg atg atg aaa gcg aga 1536
Ala Ala Ala Ala Gly Ala Glu Ser Thr Lys Thr Met Met Lys Ala Arg
500 505 510
gcc ggc cgc gcc tcc tat gtc ggc gaa agg gat ctg gca ggt gtc gct 1584
Ala Gly Arg Ala Ser Tyr Val Gly Glu Arg Asp Leu Ala Gly Val Ala
515 520 525
gat ccc ggc gcc gtc gcg gtt gcc ggc gcg ttc ggt gtg gcg gca agc 1632
Asp Pro Gly Ala ValAla Val Ala Gly Ala Phe Gly Val Ala Ala Ser
530 535 540
ctc gcc tga 1641
Leu Ala
545
<210>57
<211>546
<212>PRT
<213>豌豆根瘤菌
<400>57
Met Lys His Phe Phe Asn Arg Arg Glu Asn Ile Val Thr Glu Ala Leu
1 5 10 15
Asp Gly Leu Leu Leu Thr Ser Ser Lys Gly Arg Leu Ala Arg Leu Asp
20 25 30
Ser Phe Pro Asp Ile Lys Val Ile Leu Arg Ala Asp Trp Asp Lys Ser
35 40 45
Lys Val Ala Ile Ile Ser Gly Gly Gly Ala Gly His Glu Pro Ser His
50 55 60
Ala Gly Phe Val Gly Lys Gly Met Leu Thr Ala Ala Val Ser Gly Glu
65 70 75 80
Ile Phe Ala Ser Pro Ser Val Asp Ala Val Leu Thr Ala Ile Arg Ala
85 90 95
Val Ala Gly Glu Lys Gly Ala Leu Leu Ile Val Lys Asn Tyr Thr Gly
100 105 110
Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Lys Ala Arg Ala Glu Gly
115 120 125
Phe Asp Val Glu Met Val Ile Val Ala Asp Asp Ile Ala Ile Pro Glu
130 135 140
Ile Asn Gln Pro Arg Gly Val Ala Gly Thr Leu Phe Val His Lys Ile
145 150 155 160
Ala Gly Tyr His Ala Glu Arg Gly Glu Asp Leu Lys Thr Val Ala Ala
165 170 175
His Ala Ala Ala Ala Ala Gly Asp Ile Val Ser Leu Gly Met Ser Leu
180 185 190
Ser Thr Cys Ser Val Pro Gly Gln Ala His Glu Ser Arg Leu Gly Glu
195 200 205
Asn Glu Gly Glu Leu Gly Leu Gly Ile His Gly Glu Pro Gly Val Glu
210 215 220
Arg Ile Ala Leu Gln Pro Val Val Asp Ile Val Ala Thr Met Val Ala
225 230 235 240
Arg Leu Ser Pro Ala Leu Arg Glu Gly Gly Asn His Ala Leu Leu Ile
245 250 255
Asn Asn Leu Gly Ala Val Pro Pro Leu Glu Met Thr Val Ile Ala Asn
260 265 270
Val Val Leu Ser Ser Ser Leu Ala Asp Arg Val Arg Leu IIe Ile Gly
275 280 285
Pro Ala Pro Met Met Thr Ala Leu Asn Met Asn Gly Phe Ser Leu Ser
290 295 300
Leu Ile Arg Leu Asp Ala Ala Arg Glu Ala Ala Leu Thr Ala Ala Val
305 310 315 320
Glu Pro His Ala Trp Met Pro Ala Val Glu Arg His Glu Ile Arg Val
325 330 335
Ile Ala Ala Pro Arg Thr Ser Ala Gly Leu Asn Gly Ala Pro Val Ala
340 345 350
Gly Asp Asn Leu Arg Asn Arg Arg Leu Ile Thr Ala Leu Cys Glu His
355 360 365
Leu Ile Ser Gln Glu Ser Glu Leu Asn Arg Leu Asp Gly Arg Val Gly
370 375 380
Asp Gly Asp Thr Gly Ser Thr Val Ala Thr Gly Ala Arg Ser Val Leu
385 390 395 400
Ala Arg Leu Asp Thr Leu Pro Leu Asp Arg Pro Ala Ala Thr Leu Ala
405 410 415
Ser Leu Gly Asp Ile Leu Gly Thr Ser Met Gly Gly Ser Ser Gly Val
420 425 430
Leu Leu Ser Ile Phe Phe Thr Ala Ala Ala Lys Ala Met Ala Asp Lys
435 440 445
Ala Asp Ile Ser Ala Ala Leu Ile Ala Gly Leu Asp Arg Met Thr Phe
450 455 460
Tyr Gly Gly Ala Glu Val Gly Asp Arg Thr Met Val Asp Ala Leu Ser
465 470 475 480
Pro Ala Leu Gln Ala Leu Ala Ser Gly Asp Val Ala Ala Ala Ala Arg
485 490 495
Ala Ala Ala Ala Gly Ala Glu Ser Thr Lys Thr Met Met Lys Ala Arg
500 505 510
Ala Gly Arg Ala Ser Tyr Val Gly Glu Arg Asp Leu Ala Gly Val Ala
515 520 525
Asp Pro Gly Ala Val Ala Val Ala Gly Ala Phe Gly Val Ala Ala Ser
530 535 540
Leu Ala
545
<210>58
<211>1701
<212>DNA
<213>黄色粘球菌(Myxococcus xanthus)
<220>
<221>CDS
<222>(1)..(1701)
<223>
<400>58
atg aag aag ctg gtc aac gcc cct cgc gcg gtg gtg cgg gag atg ctg 48
Met Lys Lys Leu Val Asn Ala Pro Arg Ala Val Val Arg Glu Met Leu
1 5 10 15
gag ggg ttg gtc tcg ctc gcc ccc ggg cag gtg ctg ctg gac ggg gag 96
Glu Gly Leu Val Ser Leu Ala Pro Gly Gln Val Leu Leu Asp Gly Glu
20 25 30
tcg gtg gtg ctc cgc gcc gac acg cct tcc gac gtc cgc gcg cgc aag 144
Ser Val Val Leu Arg Ala Asp Thr Pro Ser Asp Val Arg Ala Arg Lys
35 40 45
gtg gct gtc atc tcc ggt ggc ggc agc ggc cat gag ccg gcg cac gcg 192
Val Ala Val Ile Ser Gly Gly Gly Ser Gly His Glu Pro Ala His Ala
50 55 60
ggc tac gtg ggc gcg ggc atg ctg gac gcg gcg gtg gcc ggt gac gtc 240
Gly Tyr Val Gly Ala Gly Met Leu Asp Ala Ala Val Ala Gly Asp Val
65 70 75 80
ttc acc tcg ccc agc acc gat gcc gtg ctg gcc gcc atc cgc gcc gtc 288
Phe Thr Ser Pro Ser Thr Asp Ala Val Leu Ala Ala Ile Arg Ala Val
85 90 95
gcg ggg ccc gcg ggc gcg ctg ctc gtc gtg aag aac tac acc ggg gac 336
Ala Gly Pro Ala Gly Ala Leu Leu Val Val Lys Asn Tyr Thr Gly Asp
100 105 110
cgg ctc aac ttc ggg ctc gcc gcc gag ctg gcg cgc gcc gag ggc atc 384
Arg Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Glu Gly Ile
115 120 125
ccc gtg gag acg gtg gtg gtg gcg gac gac gtg tcc ctg cac gac acg 432
Pro Val Glu Thr Val Val Val Ala Asp Asp Val Ser Leu His Asp Thr
130 135 140
gtg gag ccc gcg cgg cgc cgg ggc atc gct ggc acg gtg ctg gtc cac 480
Val Glu Pro Ala Arg Arg Arg Gly Ile Ala Gly Thr Val Leu Val His
145 150 155 160
aag gtc gcg ggc gcg gcg gcc gag gcg ggc gcg gcg ctc cag gac gtc 528
Lys Val Ala Gly Ala Ala Ala Glu Ala Gly Ala Ala Leu Gln Asp Val
165 170 175
ctc cgc gag gcc acc gcg gcg gcg gag gtg ctg ggc acc atg ggc gtg 576
Leu Arg Glu Ala Thr Ala Ala Ala Glu Val Leu Gly Thr Met Gly Val
180 185 190
gcc ctg ggg ccc tgc acc gtg ccc gcg gcg ggc aag ccg ggc ttc acg 624
Ala Leu Gly Pro Cys Thr Val Pro Ala Ala Gly Lys Pro Gly Phe Thr
195 200 205
ctg gag gag gac gaa atc gag ctg ggc ctg ggc atc cac ggc gag cag 672
Leu Glu Glu Asp Glu Ile Glu Leu Gly Leu Gly Ile His Gly Glu Gln
210 215 220
ggc gtg cgg cgc gtg ccg atg cag acg gcg gac agc ctg gtg gac acg 720
Gly Val Arg Arg Val Pro Met Gln Thr Ala Asp Ser Leu Val Asp Thr
225 230 235 240
ctg ctc acc acc atc gtc gag gac cgg cgc atc acc tcg gga gac agg 768
Leu Leu Thr Thr Ile Val Glu Asp Arg Arg Ile Thr Ser Gly Asp Arg
245 250 255
gtg gtg ctg gtg gtc aac gga ttg ggc ggc acg ccg ccc atg gag ctg 816
Val Val Leu Val Val Asn Gly Leu Gly Gly Thr Pro Pro Met Glu Leu
260 265 270
gcc atc gtc gcc cgg cgc gca ctg gcc gct ctg cgt cag ggc ggc atc 864
Ala Ile Val Ala Arg Arg Ala Leu Ala Ala Leu Arg Gln Gly Gly Ile
275 280 285
cgc gtg gag cgc gcg tgg agc ggg acg ttc ctc tcc gcg ctg gag atg 912
Arg Val Glu Arg Ala Trp Ser Gly Thr Phe Leu Ser Ala Leu Glu Met
290 295 300
ccc ggc tgc tcg ttg acg ctg ctg aag gtg gac gac gcg cgg ctg gcc 960
Pro Gly Cys Ser Leu Thr Leu Leu Lys Val Asp Asp Ala Arg Leu Ala
305 310 315 320
cgc ctg gat gcg gcg gtg gat gcg ccc gcg tgg ccc ggc gcg gga cgg 1008
Arg Leu Asp Ala Ala Val Asp Ala Pro Ala Trp Pro Gly Ala Gly Arg
325 330 335
ctg ccg aag gag ccg ggg gtg tac cgg cct tcg tcc acg gcg tct cca 1056
Leu Pro Lys Glu Pro Gly Val Tyr Arg Pro Ser Ser Thr Ala Ser Pro
340 345 350
gca tcg ctt ccg gcg gag gcg ccg caa ccg ggg atg gac cgc ttc cgg 1104
Ala Ser Leu Pro Ala Glu Ala Pro Gln Pro Gly Met Asp Arg Phe Arg
355 360 365
aag gcc gcc ttg cgg gtg gcg gac gca ttc gag cag tcg gag ccc cgg 1152
Lys Ala Ala Leu Arg Val Ala Asp Ala Phe Glu Gln Ser Glu Pro Arg
370 375 380
ctg acc gcg ctc gat agc gcc gcg ggc gac ggt gac ctg ggc ctc agt 1200
Leu Thr Ala Leu Asp Ser Ala Ala Gly Asp Gly Asp Leu Gly Leu Ser
385 390 395 400
ctg gtg cgt ggc gcc gag gcg att cgc gct ctt ccg gag gac gcg tgg 1248
Leu Val Arg Gly Ala Glu Ala Ile Arg Ala Leu Pro Glu Asp Ala Trp
405 410 415
acg agc ccc gcg cgt gcg ctg acg gcc att ggc aat gcc ttg cgg cgc 1296
Thr Ser Pro Ala Arg Ala Leu Thr Ala Ile Gly Asn Ala Leu Arg Arg
420 425 430
agc att ggc ggc agc tcg ggg ccc ttc tac gcg acg gcg ctg ctg cgc 1344
Ser Ile Gly Gly Ser Ser Gly Pro Phe Tyr Ala Thr Ala Leu Leu Arg
435 440 445
gcc gcg cgc agg ctg gcg gaa ggg ccc gtg gat gcc gcc gca tgg gcc 1392
Ala Ala Arg Arg Leu Ala Glu Gly Pro Val Asp Ala Ala Ala Trp Ala
450 455 460
gag gcc ttc gac ctc gcc gtc acc gcc gta tcg gag ctg ggc ggc gcg 1440
Glu Ala Phe Asp Leu Ala Val Thr Ala Val Ser Glu Leu Gly Gly Ala
465 470 475 480
cgg cct ggg gac cgc acc atg ctc gat gca ctc cgg ccc gcc gcc gac 1488
Arg Pro Gly Asp Arg Thr Met Leu Asp Ala Leu Arg Pro Ala Ala Asp
485 490 495
gcc ttc gcg aag gcg gtg cgt tgc ggg cag ggg gct cgc gag gcc tgg 1536
Ala Phe Ala Lys Ala Val Arg Cys Gly Gln Gly Ala Arg Glu Ala Trp
500 505 510
gcc tcg gcg gtg cac gcg gcc gag gcg gga gag gag gcg acg tcc cgg 1584
Ala Ser Ala Val His Ala Ala Glu Ala Gly Glu Glu Ala Thr Ser Arg
515 520 525
atg cag ccg cgc ctg gga cgc gcc agc tac ctg ggt gcg cgc gcc gtc 1632
Met Gln Pro Arg Leu Gly Arg Ala Ser Tyr Leu Gly Ala Arg Ala Val
530 535 540
ggt gtg ccg gac gcg ggc gcc gcg gcc gtg gtg gtg tgg atg aag gcg 1680
Gly Val Pro Asp Ala Gly Ala Ala Ala Val Val Val Trp Met Lys Ala
545 550 555 560
ctc acg cct ggc atc ggc tga 1701
Leu Thr Pro Gly Ile Gly
565
<210>59
<211>566
<212>PRT
<213>黄色粘球菌
<400>59
Met Lys Lys Leu Val Asn Ala Pro Arg Ala Val Val Arg Glu Met Leu
1 5 10 15
Glu Gly Leu Val Ser Leu Ala Pro Gly Gln Val Leu Leu Asp Gly Glu
20 25 30
Ser Val Val Leu Arg Ala Asp Thr Pro Ser Asp Val Arg Ala Arg Lys
35 40 45
Val Ala Val Ile Ser Gly Gly Gly Ser Gly His Glu Pro Ala His Ala
50 55 60
Gly Tyr Val Gly Ala Gly Met Leu Asp Ala Ala Val Ala Gly Asp Val
65 70 75 80
Phe Thr Ser Pro Ser Thr Asp Ala Val Leu Ala Ala Ile Arg Ala Val
85 90 95
Ala Gly Pro Ala Gly Ala Leu Leu Val Val Lys Asn Tyr Thr Gly Asp
100 105 110
Arg Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Glu Gly Ile
115 120 125
Pro Val Glu Thr Val Val Val Ala Asp Asp Val Ser Leu His Asp Thr
130 135 140
Val Glu Pro Ala Arg Arg Arg Gly Ile Ala Gly Thr Val Leu Val His
145 150 155 160
Lys Val Ala Gly Ala Ala Ala Glu Ala Gly Ala Ala Leu Gln Asp Val
165 170 175
Leu Arg Glu Ala Thr Ala Ala Ala Glu Val Leu Gly Thr Met Gly Val
180 185 190
Ala Leu Gly Pro Cys Thr Val Pro Ala Ala Gly Lys Pro Gly Phe Thr
195 200 205
Leu Glu Glu Asp Glu Ile Glu Leu Gly Leu Gly Ile His Gly Glu Gln
210 215 220
Gly Val Arg Arg Val Pro Met Gln Thr Ala Asp Ser Leu Val Asp Thr
225 230 235 240
Leu Leu Thr Thr Ile Val Glu Asp Arg Arg Ile Thr Ser Gly Asp Arg
245 250 255
Val Val Leu Val Val Asn Gly Leu Gly Gly Thr Pro Pro Met Glu Leu
260 265 270
Ala Ile Val Ala Arg Arg Ala Leu Ala Ala Leu Arg Gln Gly Gly Ile
275 280 285
Arg Val Glu Arg Ala Trp Ser Gly Thr Phe Leu Ser Ala Leu Glu Met
290 295 300
Pro Gly Cys Ser Leu Thr Leu Leu Lys Val Asp Asp Ala Arg Leu Ala
305 310 315 320
Arg Leu Asp Ala Ala Val Asp Ala Pro Ala Trp Pro Gly Ala Gly Arg
325 330 335
Leu Pro Lys Glu Pro Gly Val Tyr Arg Pro Ser Ser Thr Ala Ser Pro
340 345 350
Ala Ser Leu Pro Ala Glu Ala Pro Gln Pro Gly Met Asp Arg Phe Arg
355 360 365
Lys Ala Ala Leu Arg Val Ala Asp Ala Phe Glu Gln Ser Glu Pro Arg
370 375 380
Leu Thr Ala Leu Asp Ser Ala Ala Gly Asp Gly Asp Leu Gly Leu Ser
385 390 395 400
Leu Val Arg Gly Ala Glu Ala Ile Arg Ala Leu Pro Glu Asp Ala Trp
405 410 415
Thr Ser Pro Ala Arg Ala Leu Thr Ala Ile Gly Asn Ala Leu Arg Arg
420 425 430
Ser Ile Gly Gly Ser Ser Gly Pro Phe Tyr Ala Thr Ala Leu Leu Arg
435 440 445
Ala Ala Arg Arg Leu Ala Glu Gly Pro Val Asp Ala Ala Ala Trp Ala
450 455 460
Glu Ala Phe Asp Leu Ala Val Thr Ala Val Ser Glu Leu Gly Gly Ala
465 470 475 480
Arg Pro Gly Asp Arg Thr Met Leu Asp Ala Leu Arg Pro Ala Ala Asp
485 490 495
Ala Phe Ala Lys Ala Val Arg Cys Gly Gln Gly Ala Arg Glu Ala Trp
500 505 510
Ala Ser Ala Val His Ala Ala Glu Ala Gly Glu Glu Ala Thr Ser Arg
515 520 525
Met Gln Pro Arg Leu Gly Arg Ala Ser Tyr Leu Gly Ala Arg Ala Val
530 535 540
Gly Val Pro Asp Ala Gly Ala Ala Ala Val Val Val Trp Met Lys Ala
545 550 555 560
Leu Thr Pro Gly Ile Gly
565
<210>60
<211>1701
<212>DNA
<213>伯克霍尔德氏菌属菌种383
<220>
<221>CDS
<222>(1)..(1701)
<223>
<400>60
atg aaa aag ctt gtc aac cgc ccg tcc gat gtc gtg cga gaa atg ctg 48
Met Lys Lys Leu Val Asn Arg Pro Ser Asp Val Val Arg Glu Met Leu
1 5 10 15
gaa ggc atc gcg cgg cag tcg ccg cat gtc gcg atc ctc ggc gac gag 96
Glu Gly Ile Ala Arg Gln Ser Pro His Val Ala Ile Leu Gly Asp Glu
20 25 30
cac gtg ctc gtc cgc cag ccg ctg ccc gag ccc gcg caa cgc ccc gtc 144
His Val Leu Val Arg Gln Pro Leu Pro Glu Pro Ala Gln Arg Pro Val
35 40 45
gcc atc ctg tcc ggt ggc ggc agc ggc cac gag ccc gcg cac ggc ggc 192
Ala Ile Leu Ser Gly Gly Gly Ser Gly His Glu Pro Ala His Gly Gly
50 55 60
tat gtc ggc gaa gga atg ctg agc gcg gcc gtc tgc ggc gaa gtg ttc 240
Tyr Val Gly Glu Gly Met Leu Ser Ala Ala Val Cys Gly Glu Val Phe
65 70 75 80
acg tcg ccg tcc aca gac gcc gtg ctc gcc gcg atc cgc gcg agc gcc 288
Thr Ser Pro Ser Thr Asp Ala Val Leu Ala Ala Ile Arg Ala Ser Ala
85 90 95
ggc ccg aac ggc gcc ttg ctg atc gtg aag aac tac acg ggc gac cgg 336
Gly Pro Asn Gly Ala Leu Leu Ile Val Lys Asn Tyr Thr Gly Asp Arg
100 105 110
ctc aat ttc ggg ctc gcc gcc gaa ctc gca cgc gcc gaa ggc att ccg 384
Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Glu Gly Ile Pro
115 120 125
gtc gag acg gtc atc gtc gcc gac gac gta tcg ctg cgc ggc cgc gtc 432
Val Glu Thr Val Ile Val Ala Asp Asp Val Ser Leu Arg Gly Arg Val
130 135 140
gag cgc ggc cag cgg cgc ggg atc gcc ggt acc gtg ctg atc cac aag 480
Glu Arg Gly Gln Arg Arg Gly Ile Ala Gly Thr Val Leu Ile His Lys
145 150 155 160
ctc gcc ggc gcg gca gcc gcg cgc ggg ctg ccg ctc gcc cgc gtc gcg 528
Leu Ala Gly Ala Ala Ala Ala Arg Gly Leu Pro Leu Ala Arg Val Ala
165 170 175
gcc atc gcg cgc gac gcg gcg gcc gaa ctc ggc acg atg ggt gtc gca 576
Ala Ile Ala Arg Asp Ala Ala Ala Glu Leu Gly Thr Met Gly Val Ala
180 185 190
ctc gac ggc tgc acg atc ccg ggc gcc gac aag tcg ggc ttc agc ctc 624
Leu Asp Gly Cys Thr Ile Pro Gly Ala Asp Lys Ser Gly Phe Ser Leu
195 200 205
ggc gat cac gag atc gaa ctc ggc ctc ggc atc cat ggc gag aaa ggc 672
Gly Asp His Glu Ile Glu Leu Gly Leu Gly Ile His Gly Glu Lys Gly
210 215 220
gtc gag cgc cgc gcg ccg ctg ccg gcc gat gcg ctt gtc gac acg ctg 720
Val Glu Arg Arg Ala Pro Leu Pro Ala Asp Ala Leu Val Asp Thr Leu
225 230 235 240
ctg tcg agc atc gcc gcc gat ctc gtg ctc gac cgc ggc gaa cgc gtt 768
Leu Ser Ser Ile Ala Ala Asp Leu Val Leu Asp Arg Gly Glu Arg Val
245 250 255
gcg ctg ttc gtc aac ggc ctc ggc gcg acg ccg gac atg gaa ctc gcg 816
Ala Leu Phe Val Asn Gly Leu Gly Ala Thr Pro Asp Met Glu Leu Ala
260 265 270
atc gtg ctg cgc gcc gcg cac gac aac ctg cac cgg cgc ggc atc gtc 864
Ile Val Leu Arg Ala Ala His Asp Asn Leu His Arg Arg Gly Ile Val
275 280 285
gtc gcg cgt gcg tgg gcc ggc acg ttc ctg tcg gcg ctg aac atg ccc 912
Val Ala Arg Ala Trp Ala Gly Thr Phe Leu Ser Ala Leu Asn Met Pro
290 295 300
ggc tgc tcg atc tcg gtg ctg cgg ctg aac gac gaa cgc gcg gtg ctg 960
Gly Cys Ser Ile Ser Val Leu Arg Leu Asn Asp Glu Arg Ala Val Leu
305 310 315 320
ctc gac gca ccg acg cag gcg cgt gca tgg cca ggc ggc ggc gcg gtg 1008
Leu Asp Ala Pro Thr Gln Ala Arg Ala Trp Pro Gly Gly Gly Ala Val
325 330 335
aat acg cag atc cgt gtg gcc tcg gcc gcc gtg cag gaa gcg ccg ttg 1056
Asn Thr Gln Ile Arg Val Ala Ser Ala Ala Val Gln Glu Ala Pro Leu
340 345 350
ccg ccg ctc gat gcg gcc ggc cgc gcg tgg gcc gcg cgc ctg caa ccg 1104
Pro Pro Leu Asp Ala Ala Gly Arg Ala Trp Ala Ala Arg Leu Gln Pro
355 360 365
gca ttg cac gcg gtc gcg caa acg ctg atc gat cac gag cag acg ctg 1152
Ala Leu His Ala Val Ala Gln Thr Leu Ile Asp His Glu Gln Thr Leu
370 375 380
acc gac ctc gat gcg gcg gcc ggc gac ggc gat ctc ggc gcg agc atg 1200
Thr Asp Leu Asp Ala Ala Ala Gly Asp Gly Asp Leu Gly Ala Ser Met
385 390 395 400
ctg cgc gcc gcg cag gcg atc ctc gca ctg ccg gaa agc gca tac ggc 1248
Leu Arg Ala Ala Gln Ala Ile Leu Ala Leu Pro Glu Ser Ala Tyr Gly
405 410 415
acg ccg gcc ggc gcg ctc tcg gcg ctc ggc gcc gcg ttg cgc cgc gcg 1296
Thr Pro Ala Gly Ala Leu Ser Ala Leu Gly Ala Ala Leu Arg Arg Ala
420 425 430
atc gcc ggc agc tcg ggg ccg ttc tat gcg acc gcg ctg ctg cgc gcg 1344
Ile Ala Gly Ser Ser Gly Pro Phe Tyr Ala Thr Ala Leu Leu Arg Ala
435 440 445
tcg cgc cgg ctg gcc gat atc gcc gag ccg tcc gca cgc gac tgg gcc 1392
Ser Arg Arg Leu Ala Asp Ile Ala Glu Pro Ser Ala Arg Asp Trp Ala
450 455 460
gcg gcg ttc cgc ggc gcg gtg gat tcg atc agc gaa ctg ggc ggc gcg 1440
Ala Ala Phe Arg Gly Ala Val Asp Ser Ile Ser Glu Leu Gly Gly Ala
465 470 475 480
cac gcc ggc gac cgg acc atg ctc gat gcg ctg gtc ccg gcc gtc gcg 1488
His Ala Gly Asp Arg Thr Met Leu Asp Ala Leu Val Pro Ala Val Ala
485 490 495
gca ttc gag cgg gcg ctc gac aac gat cgc gat ccc gcc ggc gca tgg 1536
Ala Phe Glu Arg Ala Leu Asp Asn Asp Arg Asp Pro Ala Gly Ala Trp
500 505 510
acg gcc gcg gtc gaa gcc gcc gag cac ggt gcg cag gaa acc gca cgc 1584
Thr Ala Ala Val Glu Ala Ala Glu His Gly Ala Gln Glu Thr Ala Arg
515 520 525
atg acg cca cgc gcc ggg cgc gcg agc tat ctc ggc gaa cgt gcg atc 1632
Met Thr Pro Arg Ala Gly Arg Ala Ser Tyr Leu Gly Glu Arg Ala Ile
530 535 540
ggc acg ccg gac ggc ggc gcg gtc gcg gtg tcg tat tgg ctg cgt gcg 1680
Gly Thr Pro Asp Gly Gly Ala Val Ala Val Ser Tyr Trp Leu Arg Ala
545 550 555 560
ttg cag gca cac atc ggg tga 1701
Leu Gln Ala His Ile Gly
565
<210>61
<211>566
<212>PRT
<213>伯克霍尔德氏菌属菌种383
<400>61
Met Lys Lys Leu Val Asn Arg Pro Ser Asp Val Val Arg Glu Met Leu
1 5 10 15
Glu Gly Ile Ala Arg Gln Ser Pro His Val Ala Ile Leu Gly Asp Glu
20 25 30
His Val Leu Val Arg Gln Pro Leu Pro Glu Pro Ala Gln Arg Pro Val
35 40 45
Ala Ile Leu Ser Gly Gly Gly Ser Gly His Glu Pro Ala His Gly Gly
50 55 60
Tyr Val Gly Glu Gly Met Leu Ser Ala Ala Val Cys Gly Glu Val Phe
65 70 75 80
Thr Ser Pro Ser Thr Asp Ala Val Leu Ala Ala Ile Arg Ala Ser Ala
85 90 95
Gly Pro Asn Gly Ala Leu Leu Ile Val Lys Asn Tyr Thr Gly Asp Arg
100 105 110
Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Glu Gly Ile Pro
115 120 125
Val Glu Thr Val Ile Val Ala Asp Asp Val Ser Leu Arg Gly Arg Val
130 135 140
Glu Arg Gly Gln Arg Arg Gly Ile Ala Gly Thr Val Leu Ile His Lys
145 150 155 160
Leu Ala Gly Ala Ala Ala Ala Arg Gly Leu Pro Leu Ala Arg Val Ala
165 170 175
Ala Ile Ala Arg Asp Ala Ala Ala Glu Leu Gly Thr Met Gly Val Ala
180 185 190
Leu Asp Gly Cys Thr Ile Pro Gly Ala Asp Lys Ser Gly Phe Ser Leu
195 200 205
Gly Asp His Glu Ile Glu Leu Gly Leu Gly Ile His Gly Glu Lys Gly
210 215 220
Val Glu Arg Arg Ala Pro Leu Pro Ala Asp Ala Leu Val Asp Thr Leu
225 230 235 240
Leu Ser Ser Ile Ala Ala Asp Leu Val Leu Asp Arg Gly Glu Arg Val
245 250 255
Ala Leu Phe Val Asn Gly Leu Gly Ala Thr Pro Asp Met Glu Leu Ala
260 265 270
Ile Val Leu Arg Ala Ala His Asp Asn Leu His Arg Arg Gly Ile Val
275 280 285
Val Ala Arg Ala Trp Ala Gly Thr Phe Leu Ser Ala Leu Asn Met Pro
290 295 300
Gly Cys Ser Ile Ser Val Leu Arg Leu Asn Asp Glu Arg Ala Val Leu
305 310 315 320
Leu Asp Ala Pro Thr Gln Ala Arg Ala Trp Pro Gly Gly Gly Ala Val
325 330 335
Asn Thr Gln Ile Arg Val Ala Ser Ala Ala Val Gln Glu Ala Pro Leu
340 345 350
Pro Pro Leu Asp Ala Ala Gly Arg Ala Trp Ala Ala Arg Leu Gln Pro
355 360 365
Ala Leu His Ala Val Ala Gln Thr Leu Ile Asp His Glu Gln Thr Leu
370 375 380
Thr Asp Leu Asp Ala Ala Ala Gly Asp Gly Asp Leu Gly Ala Ser Met
385 390 395 400
Leu Arg Ala Ala Gln Ala Ile Leu Ala Leu Pro Glu Ser Ala Tyr Gly
405 410 415
Thr Pro Ala Gly Ala Leu Ser Ala Leu Gly Ala Ala Leu Arg Arg Ala
420 425 430
Ile Ala Gly Ser Ser Gly Pro Phe Tyr Ala Thr Ala Leu Leu Arg Ala
435 440 445
Ser Arg Arg Leu Ala Asp Ile Ala Glu Pro Ser Ala Arg Asp Trp Ala
450 455 460
Ala Ala Phe Arg Gly Ala Val Asp Ser Ile Ser Glu Leu Gly Gly Ala
465 470 475 480
His Ala Gly Asp Arg Thr Met Leu Asp Ala Leu Val Pro Ala Val Ala
485 490 495
Ala Phe Glu Arg Ala Leu Asp Asn Asp Arg Asp Pro Ala Gly Ala Trp
500 505 510
Thr Ala Ala Val Glu Ala Ala Glu His Gly Ala Gln Glu Thr Ala Arg
515 520 525
Met Thr Pro Arg Ala Gly Arg Ala Ser Tyr Leu Gly Glu Arg Ala Ile
530 535 540
Gly Thr Pro Asp Gly Gly Ala Val Ala Val Ser Tyr Trp Leu Arg Ala
545 550 555 560
Leu Gln Ala His Ile Gly
565
<210>62
<211>1704
<212>DNA
<213>泰国伯克霍尔德氏菌(Burkholderia thailandensis)E264
<220>
<221>CDS
<222>(1)..(1704)
<223>
<400>62
atg aag aag ctc gtc aac cac ccg tcc gac gtc gtg cgc gaa atg ctg 48
Met Lys Lys Leu Val Asn His Pro Ser Asp Val Val Arg Glu Met Leu
1 5 10 15
gag ggc atc gcg cgg cag tcg ccg cat gtc gcg atg ctc ggc gac gaa 96
Glu Gly Ile Ala Arg Gln Ser Pro His Val Ala Met Leu Gly Asp Glu
20 25 30
cac gtg ctg atc cgg cgc ccc ttg ccg gag ccg gcg cgg cgt gcg gtc 144
His Val Leu Ile Arg Arg Pro Leu Pro Glu Pro Ala Arg Arg Ala Val
35 40 45
gcg atc atc tcc ggc ggc ggc agc ggc cac gag ccg gcg cac ggc ggc 192
Ala Ile Ile Ser Gly Gly Gly Ser Gly His Glu Pro Ala His Gly Gly
50 55 60
tac gtc ggc gcg ggg atg ctg agc gcg gcc gtg tgc ggc gag gtc ttc 240
Tyr Val Gly Ala Gly Met Leu Ser Ala Ala Val Cys Gly Glu Val Phe
65 70 75 80
acg tcg ccg ccc gcc gat gcg gtg ctc gcc gcg att cgc gcg acc gcg 288
Thr Ser Pro Pro Ala Asp Ala Val Leu Ala Ala Ile Arg Ala Thr Ala
85 90 95
ggc cag aac ggc gcg ctc ctc atc gtg aag aac tac acg ggc gat cgc 336
Gly Gln Asn Gly Ala Leu Leu Ile Val Lys Asn Tyr Thr Gly Asp Arg
100 105 110
ctc aat ttc ggg ctc gcg gcc gag ctc gcg cgc gcg cag ggc att ccg 384
Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Gln Gly Ile Pro
115 120 125
gtc gag atc gtc gtc gtc gcg gac gac gtg tcg ctg cgc gaa ctc acc 432
Val Glu Ile Val Val Val Ala Asp Asp Val Ser Leu Arg Glu Leu Thr
130 135 140
gag cgc ggg cgc cgc cgc ggc atc gcc ggc acc gtg ctc gtg cac aag 480
Glu Arg Gly Arg Arg Arg Gly Ile Ala Gly Thr Val Leu Val His Lys
145 150 155 160
ctc gcc ggc gcg gcc gcc gag cgc ggc ctc gcg ctg cgg gag gtg gcc 528
Leu Ala Gly Ala Ala Ala Glu Arg Gly Leu Ala Leu Arg Glu Val Ala
165 170 175
gcc gtc gcg agc gag gcg gcg gcg aat ctc ggc acg atg ggc gtc gca 576
Ala Val Ala Ser Glu Ala Ala Ala Asn Leu Gly Thr Met Gly Val Ala
180 185 190
ctc gac ggc tgc acg att ccg ggc gcc ggg caa tcg ggc ttc cgc ctc 624
Leu Asp Gly Cys Thr Ile Pro Gly Ala Gly Gln Ser Gly Phe Arg Leu
195 200 205
gcc gat cac gag atc gag ctc gga ttg ggc att cac ggc gaa aag ggc 672
Ala Asp His Glu Ile Glu Leu Gly Leu Gly Ile His Gly Glu Lys Gly
2l0 215 220
gtg cag cgc acg gcg ccg atg ccg gcc gac gcg ctg tcg gaa acg ctc 720
Val Gln Arg Thr Ala Pro Met Pro Ala Asp Ala Leu Ser Glu Thr Leu
225 230 235 240
gtg gcg acg atc gtc gac gat cag gcg atc gcg cgc ggc gat cgg gtc 768
Val Ala Thr Ile Val Asp Asp Gln Ala Ile Ala Arg Gly Asp Arg Val
245 250 255
gcg ctt ctg gtg aac ggg ctc ggc gcg acg ccg gac atg gag ctc ggc 816
Ala Leu Leu Val Asn Gly Leu Gly Ala Thr Pro Asp Met Glu Leu Gly
260 265 270
atc gtg ctg cgc gcg gcg tac gac agc ctg agc cgg cgt ggc gtc gag 864
Ile Val Leu Arg Ala Ala Tyr Asp Ser Leu Ser Arg Arg Gly Val Glu
275 280 285
gtg gcg cgc gcg tgg gcg ggc acg ttc ctg tcc gcg ctc gac atg ccc 912
Val Ala Arg Ala Trp Ala Gly Thr Phe Leu Ser Ala Leu Asp Met Pro
290 295 300
ggc tgc tcg att tcg ctg ctc aag ctg aac gat cgc atg ctc gaa ctg 960
Gly Cys Ser Ile Ser Leu Leu Lys Leu Asn Asp Arg Met Leu Glu Leu
305 310 315 320
ctc gac gcg ccg acg caa gcg cga gcg tgg ccg ggc ggc ggc gcg gtg 1008
Leu Asp Ala Pro Thr Gln Ala Arg Ala Trp Pro Gly Gly Gly Ala Val
325 330 335
aac cgg gac att cgc gtg gcc gcc gcc ggg acc ggc gca gga gac gga 1056
Asn Arg Asp Ile Arg Val Ala Ala Ala Gly Thr Gly Ala Gly Asp Gly
340 345 350
cag ccg gaa tgg gcg acg gcc ggc gcg gcg ggt tcc gac gga ctg cgg 1104
Gln Pro Glu Trp Ala Thr Ala Gly Ala Ala Gly Ser Asp Gly Leu Arg
355 360 365
ccc gcg ctg cat gcg gtt gcc gcc gcg ctg atc gac agc gag ccc gtg 1152
Pro Ala Leu His Ala Val Ala Ala Ala Leu Ile Asp Ser Glu Pro Val
370 375 380
ttg acc gag ctc gat tcc gtc gcc ggc gac ggc gat ctg ggc gcg agc 1200
Leu Thr Glu Leu Asp Ser Val Ala Gly Asp Gly Asp Leu Gly Ala Ser
385 390 395 400
atg cgc cgc gcg gcg aac gcg atg ctc gcg ctg ccc gcc gat gcg tat 1248
Met Arg Arg Ala Ala Asn Ala Met Leu Ala Leu Pro Ala Asp Ala Tyr
405 410 415
cga ggg ccg gcg aac ctg ctc gcc gcg ctg ggc atg gcg ctg cgc cgg 1296
Arg Gly Pro Ala Asn Leu Leu Ala Ala Leu Gly Met Ala Leu Arg Arg
420 425 430
gcg atc gcg ggc agc tcc ggg ccg ttt tac gca acg gcg ctg gtg cgc 1344
Ala Ile Ala Gly Ser Ser Gly Pro Phe Tyr Ala Thr Ala Leu Val Arg
435 440 445
gcg ggc cgc cgg ctc gcc gat gca tcg gcg ccg acc gcg cgc gac tgg 1392
Ala Gly Arg Arg Leu Ala Asp Ala Ser Ala Pro Thr Ala Arg Asp Trp
450 455 460
gcg agc gcg ttc cgg agc ggt gtc gac gcg atc ggc gat ctg ggc ggc 1440
Ala Ser Ala Phe Arg Ser Gly Val Asp Ala Ile Gly Asp Leu Gly Gly
465 470 475 480
gcg aag ccc gga gac cgc acg atg ctc gac gcc ttg gtg ccc gcc gtc 1488
Ala Lys Pro Gly Asp Arg Thr Met Leu Asp Ala Leu Val Pro Ala Val
485 490 495
gat gcg ttc gag cac gcg ctg tcg gcg ggc ggg agc gcg agc gat gcg 1536
Asp Ala Phe Glu His Ala Leu Ser Ala Gly Gly Ser Ala Ser Asp Ala
500 505 510
tgg gcg gcg gcg gtg cgc gcc gcg gag gca ggc gcg gcg aaa acc gcg 1584
Trp Ala Ala Ala Val Arg Ala Ala Glu Ala Gly Ala Ala Lys Thr Ala
515 520 525
ggc atg acg ccg cgc gcg ggg cgc gcg agc tat ctg ggc gag cgc gcc 1632
Gly Met Thr Pro Arg Ala Gly Arg Ala Ser Tyr Leu Gly Glu Arg Ala
530 535 540
gtg ggc tcg ccc gac ggc ggc gcg gtg gcc gtg gcg tgc tgg atg cgc 1680
Val Gly Ser Pro Asp Gly Gly Ala Val Ala Val Ala Cys Trp Met Arg
545 550 555 560
gcg ttg cag ccg cac gtc gcg tag 1704
Ala Leu Gln Pro His Val Ala
565
<210>63
<211>567
<212>PRT
<213>泰国伯克霍尔德氏菌E264
<400>63
Met Lys Lys Leu Val Asn His Pro Ser Asp Val Val Arg Glu Met Leu
1 5 10 15
Glu Gly Ile Ala Arg Gln Ser Pro His Val Ala Met Leu Gly Asp Glu
20 25 30
His Val Leu Ile Arg Arg Pro Leu Pro Glu Pro Ala Arg Arg Ala Val
35 40 45
Ala Ile Ile Ser Gly Gly Gly Ser Gly His Glu Pro Ala His Gly Gly
50 55 60
Tyr Val Gly Ala Gly Met Leu Ser Ala Ala Val Cys Gly Glu Val Phe
65 70 75 80
Thr Ser Pro Pro Ala Asp Ala Val Leu Ala Ala Ile Arg Ala Thr Ala
85 90 95
Gly Gln Asn Gly Ala Leu Leu Ile Val Lys Asn Tyr Thr Gly Asp Arg
100 105 110
Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Gln Gly Ile Pro
115 120 125
Val Glu Ile Val Val Val Ala Asp Asp Val Ser Leu Arg Glu Leu Thr
130 135 140
Glu Arg Gly Arg Arg Arg Gly Ile Ala Gly Thr Val Leu Val His Lys
145 150 155 160
Leu Ala Gly Ala Ala Ala Glu Arg Gly Leu Ala Leu Arg Glu Val Ala
165 170 175
Ala Val Ala Ser Glu Ala Ala Ala Asn Leu Gly Thr Met Gly Val Ala
180 185 190
Leu Asp Gly Cys Thr Ile Pro Gly Ala Gly Gln Ser Gly Phe Arg Leu
195 200 205
Ala Asp His Glu Ile Glu Leu Gly Leu Gly Ile His Gly Glu Lys Gly
210 215 220
Val Gln Arg Thr Ala Pro Met Pro Ala Asp Ala Leu Ser Glu Thr Leu
225 230 235 240
Val Ala Thr Ile Val Asp Asp Gln Ala Ile Ala Arg Gly Asp Arg Val
245 250 255
Ala Leu Leu Val Asn Gly Leu Gly Ala Thr Pro Asp Met Glu Leu Gly
260 265 270
Ile Val Leu Arg Ala Ala Tyr Asp Ser Leu Ser Arg Arg Gly Val Glu
275 280 285
Val Ala Arg Ala Trp Ala Gly Thr Phe Leu Ser Ala Leu Asp Met Pro
290 295 300
Gly Cys Ser Ile Ser Leu Leu Lys Leu Asn Asp Arg Met Leu Glu Leu
305 310 315 320
Leu Asp Ala Pro Thr Gln Ala Arg Ala Trp Pro Gly Gly Gly Ala Val
325 330 335
Asn Arg Asp Ile Arg Val Ala Ala Ala Gly Thr Gly Ala Gly Asp Gly
340 345 350
Gln Pro Glu Trp Ala Thr Ala Gly Ala Ala Gly Ser Asp Gly Leu Arg
355 360 365
Pro Ala Leu His Ala Val Ala Ala Ala Leu Ile Asp Ser Glu Pro Val
370 375 380
Leu Thr Glu Leu Asp Ser Val Ala Gly Asp Gly Asp Leu Gly Ala Ser
385 390 395 400
Met Arg Arg Ala Ala Asn Ala Met Leu Ala Leu Pro Ala Asp Ala Tyr
405 410 415
Arg Gly Pro Ala Asn Leu Leu Ala Ala Leu Gly Met Ala Leu Arg Arg
420 425 430
Ala Ile Ala Gly Ser Ser Gly Pro Phe Tyr Ala Thr Ala Leu Val Arg
435 440 445
Ala Gly Arg Arg Leu Ala Asp Ala Ser Ala Pro Thr Ala Arg Asp Trp
450 455 460
Ala Ser Ala Phe Arg Ser Gly Val Asp Ala Ile Gly Asp Leu Gly Gly
465 470 475 480
Ala Lys Pro Gly Asp Arg Thr Met Leu Asp Ala Leu Val Pro Ala Val
485 490 495
Asp Ala Phe Glu His Ala Leu Ser Ala Gly Gly Ser Ala Ser Asp Ala
500 505 510
Trp Ala Ala Ala Val Arg Ala Ala Glu Ala Gly Ala Ala Lys Thr Ala
515 520 525
Gly Met Thr Pro Arg Ala Gly Arg Ala Ser Tyr Leu Gly Glu Arg Ala
530 535 540
Val Gly Ser Pro Asp Gly Gly Ala Val Ala Val Ala Cys Trp Met Arg
545 550 555 560
Ala Leu Gln Pro His Val Ala
565
<210>64
<211>1851
<212>DNA
<213>多食伯克霍尔德氏菌(Burkholderia multivorans)
<220>
<221>CDS
<222>(1)..(1851)
<223>
<400>64
atg tat cgt ggc acc gaa gcg cgc acc ggc gag atc cat ggc act gct 48
Met Tyr Arg Gly Thr Glu Ala Arg Thr Gly Glu Ile His Gly Thr Ala
1 5 10 15
cca cgc cac cgc cgc cac ggc gcg cgc att ccc gca acc cga cag ccc 96
Pro Arg His Arg Arg His Gly Ala Arg Ile Pro Ala Thr Arg Gln Pro
20 25 30
gag gcg ccg gat cgg ttc ggc ccg cgg gct ttc atc tgc ccg gaa acc 144
Glu Ala Pro Asp Arg Phe Gly Pro Arg Ala Phe Ile Cys Pro Glu Thr
35 40 45
gtc gcc atg aaa aaa ctc gtg aac cgt ccg tcc gac gtc gtg cgt gaa 192
Val Ala Met Lys Lys Leu Val Asn Arg Pro Ser Asp Val Val Arg Glu
50 55 60
atg ctc gaa ggc att gcg cga cag tcg ccg cat ctc gcg atg ctc ggc 240
Met Leu Glu Gly Ile Ala Arg Gln Ser Pro His Leu Ala Met Leu Gly
65 70 75 80
gac gag cac gtg ctc gtc cgc cgc ccg ctg ccc gaa ccg tcg cag cgc 288
Asp Glu His Val Leu Val Arg Arg Pro Leu Pro Glu Pro Ser Gln Arg
85 90 95
acg gtt gcg gtg ctg tcg ggc ggc ggc agc ggg cac gag cct gcg cac 336
Thr Val Ala Val Leu Ser Gly Gly Gly Ser Gly His Glu Pro Ala His
100 105 110
ggc ggc tat gtc ggc gac gga atg ctc agt gcg gcc gtg tgc ggc gaa 384
Gly Gly Tyr Val Gly Asp Gly Met Leu Ser Ala Ala Val Cys Gly Glu
115 120 125
gtg ttc acg tcg ccg tcc acc gac gcg gtg ctc gcc gcg atc cgc gcg 432
Val Phe Thr Ser Pro Ser Thr Asp Ala Val Leu Ala Ala Ile Arg Ala
130 135 140
acg gcc ggc ccg aac ggc gcg ctg ctc gtc gtg aag aac tac acc ggc 480
Thr Ala Gly Pro Asn Gly Ala Leu Leu Val Val Lys Asn Tyr Thr Gly
145 150 155 160
gac cgg ctc aac ttc ggt ctg gca gcc gaa ctc gcg cgc gca gaa ggc 528
Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Glu Gly
165 170 175
att ccg gtc gag acc gtg atc gtc gcg gac gac gta tcg ctg cgc ggc 576
Ile Pro Val Glu Thr Val Ile Val Ala Asp Asp Val Ser Leu Arg Gly
180 185 190
cgc gtc gag cgc gga cag cgg cgc ggc atc gcg ggc acc gtg ctg atc 624
Arg Val Glu Arg Gly Gln Arg Arg Gly Ile Ala Gly Thr Val Leu Ile
195 200 205
cac aag ctc gcg ggc gcg gcg gcc gcg cgc ggg ctg tcg ctg ccg cgc 672
His Lys Leu Ala Gly Ala Ala Ala Ala Arg Gly Leu Ser Leu Pro Arg
210 215 220
gtc gcg gcg atc gcg cgc gat gcg gcc gcc gat ctc ggc acg atg ggc 720
Val Ala Ala Ile Ala Arg Asp Ala Ala Ala Asp Leu Gly Thr Met Gly
225 230 235 240
gtc gcg ctc gac ggc tgt acg ctg ccg ggc gcc gac cag tcc gga ttc 768
Val Ala Leu Asp Gly Cys Thr Leu Pro Gly Ala Asp Gln Ser Gly Phe
245 250 255
agc ctc gcc gac gac gaa atc gag ctc ggt ctc ggc att cat ggc gaa 816
Ser Leu Ala Asp Asp Glu Ile Glu Leu Gly Leu Gly Ile His Gly Glu
260 265 270
aaa ggc gtc gaa cgc acg gcg ccg ctg ccg gcc gac gcg ctc gcc gat 864
Lys Gly Val Glu Arg Thr Ala Pro Leu Pro Ala Asp Ala Leu Ala Asp
275 280 285
acg ctg ctg tcc ggg atc gtc gcc gac ctc gtg ctc gat cgc ggc gaa 912
Thr Leu Leu Ser Gly Ile Val Ala Asp Leu Val Leu Asp Arg Gly Glu
290 295 300
cgc gtc gcg ctg ctc gtc aac ggt ctc ggc gcg acg ccc gac atg gag 960
Arg Val Ala Leu Leu Val Asn Gly Leu Gly Ala Thr Pro Asp Met Glu
305 310 315 320
ctt gcg atc gtg ctg cgc gcc gcc tac gag aac ctg agc cgt cgc ggc 1008
Leu AlaIle Val Leu Arg Ala Ala Tyr Glu Asn Leu Ser Arg Arg Gly
325 330 335
atc gcg gtc gag cgc gcg tgg gcc ggt acg ttc ctg tcg gcg ctg aac 1056
Ile Ala Val Glu Arg Ala Trp Ala Gly Thr Phe Leu Ser Ala Leu Asn
340 345 350
atg ccc ggc tgt tcg atc tcg gtg ctg cgg ctc gac gac gag cgg gcg 1104
Met Pro Gly Cys Ser Ile Ser Val Leu Arg Leu Asp Asp Glu Arg Ala
355 360 365
gcg ctg ctc gac gca ccg acg caa gcg cgc gcg tgg ccc ggc ggc ggc 1152
Ala Leu Leu Asp Ala Pro Thr Gln Ala Arg Ala Trp Pro Gly Gly Gly
370 375 380
gcc gtc aac gca cag atc cgc atc gcc gcc gcg gcg ccg cag gaa ccg 1200
Ala Val Asn Ala Gln Ile ArgIle Ala Ala Ala Ala Pro Gln Glu Pro
385 390 395 400
tcg ccg ccg ccg ctc gac gcg gcg ggc cgc gtg tgg gcc gag cgg ctg 1248
Ser Pro Pro Pro Leu Asp Ala Ala Gly Arg Val Trp Ala Glu Arg Leu
405 410 415
cgc ccg gcg ctg cac gcg gtc gcg cac acg ctg atc gac cac gaa gca 1296
Arg Pro Ala Leu His Ala Val Ala His Thr Leu Ile Asp His Glu Ala
420 425 430
acg ctg acc gag ctc gat gcc gcc gcc ggc gac ggc gac ctc ggc gcg 1344
Thr Leu Thr Glu Leu Asp Ala Ala Ala Gly Asp Gly Asp Leu Gly Ala
435 440 445
agc atg cgt cgt gcc gcg gac gcg atg ctc gcg ttg ccg gaa acc gcg 1392
Ser Met Arg Arg Ala Ala Asp Ala Met Leu Ala Leu Pro Glu Thr Ala
450 455 460
tat gcg acg ccg gcc ggt gcg ctc gcc gcg ctc ggc gcc gcg ctg cgc 1440
Tyr Ala Thr Pro Ala Gly Ala Leu Ala Ala Leu Gly Ala Ala Leu Arg
465 470 475 480
cgt gcg atc gcg ggc agc tcg ggc ccc ttc tac gcg acc gcg ctg ctg 1488
Arg Ala Ile Ala Gly Ser Ser Gly Pro Phe Tyr Ala Thr Ala Leu Leu
485 490 495
cgc gca tcg cgg cgc ctg gcc ggc ctc gcg cag ccg tcg gca cgc gac 1536
Arg Ala Ser Arg Arg Leu Ala Gly Leu Ala Gln Pro Ser Ala Arg Asp
500 505 510
tgg gcc gcg gca ttg cgc agc gca gcc gat gcg atc gcc gaa ctc ggc 1584
Trp Ala Ala Ala Leu Arg Ser Ala Ala Asp Ala Ile Ala Glu Leu Gly
515 520 525
ggc gcc cgt gcc ggt gac aga acg atg ctc gac gcg ctg gtg ccg gcc 1632
Gly Ala Arg Ala Gly Asp Arg Thr Met Leu Asp Ala Leu Val Pro Ala
530 535 540
gcc gcc gcg ttc gac cgt gca ctc gac gac ggt cgc gac agc gcc ggc 1680
Ala Ala Ala Phe Asp Arg Ala Leu Asp Asp Gly Arg Asp Ser Ala Gly
545 550 555 560
gcg tgg gcg gcc gcc gtc gac gcc gcc gag cgc ggt gcg cag gac acc 1728
Ala Trp Ala Ala Ala Val Asp Ala Ala Glu Arg Gly Ala Gln Asp Thr
565 570 575
gcg cgc atg acg ccg cgc gcg ggc cgt gcg agc tat ctc ggc gaa cgc 1776
Ala Arg Met Thr Pro Arg Ala Gly Arg Ala Ser Tyr Leu Gly Glu Arg
580 585 590
gcg atc ggc acg ccc gac ggc ggc gcc atc gcc gtc gcg tac tgg ctg 1824
Ala Ile Gly Thr Pro Asp Gly Gly Ala Ile Ala Val Ala Tyr Trp Leu
595 600 605
cgc gca ctg ctg ccg cac gtc cga taa 1851
Arg Ala Leu Leu Pro His Val Arg
610 615
<210>65
<211>616
<212>PRT
<213>多食伯克霍尔德氏菌
<400>65
Met Tyr Arg Gly Thr Glu Ala Arg Thr Gly Glu Ile His Gly Thr Ala
1 5 10 15
Pro Arg His Arg Arg His Gly Ala Arg Ile Pro Ala Thr Arg Gln Pro
20 25 30
Glu Ala Pro Asp Arg Phe Gly Pro Arg Ala Phe Ile Cys Pro Glu Thr
35 40 45
Val Ala Met Lys Lys Leu Val Asn Arg Pro Ser Asp Val Val Arg Glu
50 55 60
Met Leu Glu Gly Ile Ala Arg Gln Ser Pro His Leu Ala Met Leu Gly
65 70 75 80
Asp Glu His Val Leu Val Arg Arg Pro Leu Pro Glu Pro Ser Gln Arg
85 90 95
Thr Val Ala Val Leu Ser Gly Gly Gly Ser Gly His Glu Pro Ala His
100 105 110
Gly Gly Tyr Val Gly Asp Gly Met Leu Ser Ala Ala Val Cys Gly Glu
115 120 125
Val Phe Thr Ser Pro Ser Thr Asp Ala Val Leu Ala Ala Ile Arg Ala
130 135 140
Thr Ala Gly Pro Asn Gly Ala Leu Leu Val Val Lys Asn Tyr Thr Gly
145 150 155 160
Asp Arg Leu Asn Phe Gly Leu Ala Ala Glu Leu Ala Arg Ala Glu Gly
165 170 175
Ile Pro Val Glu Thr Val Ile Val Ala Asp Asp Val Ser Leu Arg Gly
180 185 190
Arg Val Glu Arg Gly Gln Arg Arg Gly Ile Ala Gly Thr Val Leu Ile
195 200 205
His Lys Leu Ala Gly Ala Ala Ala Ala Arg Gly Leu Ser Leu Pro Arg
210 215 220
Val Ala Ala Ile Ala Arg Asp Ala Ala Ala Asp Leu Gly Thr Met Gly
225 230 235 240
Val Ala Leu Asp Gly Cys Thr Leu Pro Gly Ala Asp Gln Ser Gly Phe
245 250 255
Ser Leu Ala Asp Asp Glu Ile Glu Leu Gly Leu Gly Ile His Gly Glu
260 265 270
Lys Gly Val Glu Arg Thr Ala Pro Leu Pro Ala Asp Ala Leu Ala Asp
275 280 285
Thr Leu Leu Ser Gly Ile Val Ala Asp Leu Val Leu Asp Arg Gly Glu
290 295 300
Arg Val Ala Leu Leu Val Asn Gly Leu Gly Ala Thr Pro Asp Met Glu
305 310 315 320
Leu Ala Ile Val Leu Arg Ala Ala Tyr Glu Asn Leu Ser Arg Arg Gly
325 330 335
Ile Ala Val Glu Arg Ala Trp Ala Gly Thr Phe Leu Ser Ala Leu Asn
340 345 350
Met Pro Gly Cys Ser Ile Ser Val Leu Arg Leu Asp Asp Glu Arg Ala
355 360 365
Ala Leu Leu Asp Ala Pro Thr Gln Ala Arg Ala Trp Pro Gly Gly Gly
370 375 380
Ala Val Asn Ala Gln Ile Arg Ile Ala Ala Ala Ala Pro Gln Glu Pro
385 390 395 400
Ser Pro Pro Pro Leu Asp Ala Ala Gly Arg Val Trp Ala Glu Arg Leu
405 410 415
Arg Pro Ala Leu His Ala Val Ala His Thr Leu Ile Asp His Glu Ala
420 425 430
Thr Leu Thr Glu Leu Asp Ala Ala Ala Gly Asp Gly Asp Leu Gly Ala
435 440 445
Ser Met Arg Arg Ala Ala Asp Ala Met Leu Ala Leu Pro Glu Thr Ala
450 455 460
Tyr Ala Thr Pro Ala Gly Ala Leu Ala Ala Leu Gly Ala Ala Leu Arg
465 470 475 480
Arg Ala lle Ala Gly Ser Ser Gly Pro Phe Tyr Ala Thr Ala Leu Leu
485 490 495
Arg Ala Ser Arg Arg Leu Ala Gly Leu Ala Gln Pro Ser Ala Arg Asp
500 505 510
Trp Ala Ala Ala Leu Arg Ser Ala Ala Asp Ala Ile Ala Glu Leu Gly
515 520 525
Gly Ala Arg Ala Gly Asp Arg Thr Met Leu Asp Ala Leu Val Pro Ala
530 535 540
Ala Ala Ala Phe Asp Arg Ala Leu Asp Asp Gly Arg Asp Ser Ala Gly
545 550 555 560
Ala Trp Ala Ala Ala Val Asp Ala Ala Glu Arg Gly Ala Gln Asp Thr
565 570 575
Ala Arg Met Thr Pro Arg Ala Gly Arg Ala Ser Tyr Leu Gly Glu Arg
580 585 590
Ala Ile Gly Thr Pro Asp Gly Gly Ala Ile Ala Val Ala Tyr Trp Leu
595 600 605
Arg Ala Leu Leu Pro His Val Arg
610 615
<210>66
<211>1920
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1920)
<223>
<400>66
atg agt ggc gct ttt aac aac gat ggt cgg ggc ata tct ccc tta att 48
Met Ser Gly Ala Phe Asn Asn Asp Gly Arg Gly Ile Ser Pro Leu Ile
1 5 10 15
gca acc tcc tgg gag cga tgc aat aag ctg atg aaa cgg gag aca tgg 96
Ala Thr Ser Trp Glu Arg Cys Asn Lys Leu Met Lys Arg Glu Thr Trp
20 25 30
aac gta cca cat cag gcc cag ggc gtg aca ttt gct tct att tat cgg 144
Asn Val Pro His Gln Ala Gln Gly Val Thr Phe Ala Ser Ile Tyr Arg
35 40 45
cgt aag aaa gcg atg ctg acg ctc ggg cag gct gcg ctg gaa gat gcc 192
Arg Lys Lys Ala Met Leu Thr Leu Gly Gln Ala Ala Leu Glu Asp Ala
50 55 60
tgg gaa tat atg gca ccg cga gag tgt gcg ctg ttt atc ctc gat gaa 240
Trp Glu Tyr Met Ala Pro Arg Glu Cys Ala Leu Phe Ile Leu Asp Glu
65 70 75 80
acc gcc tgc att ctc agc cgt aat ggc gat ccg caa acc ttg cag cag 288
Thr Ala Cys Ile Leu Ser Arg Asn Gly Asp Pro Gln Thr Leu Gln Gln
85 90 95
cta agt gca ctg gga ttc aat gac ggc acg tat tgc gcc gag gga att 336
Leu Ser Ala Leu Gly Phe Asn Asp Gly Thr Tyr Cys Ala Glu Gly Ile
100 105 110
att ggt act tgt gcg cta tcg tta gcg gct atc tct ggt cag gcc gtg 384
Ile Gly Thr Cys Ala Leu Ser Leu Ala Ala Ile Ser Gly Gln Ala Val
115 120 125
aaa acg atg gcc gat caa cat ttc aaa cag gta ctc tgg aac tgg gcc 432
Lys Thr Met Ala Asp Gln His Phe Lys Gln Val Leu Trp Asn Trp Ala
130 135 140
ttt tgt gca acg ccg ttg ttt gac agc aag ggc cga ttg acg gga aca 480
Phe Cys Ala Thr Pro Leu Phe Asp Ser Lys Gly Arg Leu Thr Gly Thr
145 150 155 160
ata gcg ctg gcg tgt ccg gtt gag caa act acc gca gct gat ttg ccg 528
Ile Ala Leu Ala Cys Pro Val Glu Gln Thr Thr Ala Ala Asp Leu Pro
165 170 175
ttg acg ttg gca atc gcc cgc gag gtc gga aat tta ctg ctg acg gac 576
Leu Thr Leu Ala Ile Ala Arg Glu Val Gly Asn Leu Leu Leu Thr Asp
180 185 190
agt ttg ctc gct gaa act aac cgt cat tta aat caa ctt aat gcc ctg 624
Ser Leu Leu Ala Glu Thr Asn Arg His Leu Asn Gln Leu Asn Ala Leu
195 200 205
tta gaa agt atg gat gat ggc gtg att agc tgg gac gag cag ggt aat 672
Leu Glu Ser Met Asp Asp Gly Val Ile Ser Trp Asp Glu Gln Gly Asn
210 215 220
ttg caa ttt att aat gcc cag gcg gcg cgg gtc ttg cgc ctt gac gcg 720
Leu Gln Phe Ile Asn Ala Gln Ala Ala Arg Val Leu Arg Leu Asp Ala
225 230 235 240
acg gca agt cag gga cgg gca atc act gaa ctc tta acg tta ccc gcc 768
Thr Ala Ser Gln Gly Arg Ala Ile Thr Glu Leu Leu Thr Leu Pro Ala
245 250 255
gta ttg caa caa gca ata aaa cag gca cat ccg ctc aaa cac gta gaa 816
Val Leu Gln Gln Ala Ile Lys Gln Ala His Pro Leu Lys His Val Glu
260 265 270
gca acc ttt gaa agc cag cac cag ttt att gat gcg gtg ata acc ctt 864
Ala Thr Phe Glu Ser Gln His Gln Phe Ile Asp Ala Val Ile Thr Leu
275 280 285
aaa ccg ata ata gaa acg cag gga acc agc ttt att ttg ttg ctc cat 912
Lys Pro Ile Ile Glu Thr Gln Gly Thr Ser Phe Ile Leu Leu Leu His
290 295 300
cct gtg gaa cag atg cgg cag ttg atg acc agt caa tta gga aaa gtc 960
Pro Val Glu Gln Met Arg Gln Leu Met Thr Ser Gln Leu Gly Lys Val
305 310 315 320
agc cat acc ttc gct cat atg cca cag gac gat ccg caa acc cgc cgc 1008
Ser His Thr Phe Ala His Met Pro Gln Asp Asp Pro Gln Thr Arg Arg
325 330 335
ttg att cat ttt ggt cgc cag gcg gcg cgc agt agc ttt cct gtc ctg 1056
Leu Ile His Phe Gly Arg Gln Ala Ala Arg Ser Ser Phe Pro Val Leu
340 345 350
ctt tgt gga gaa gag ggc gtg ggc aag gca ctg cta agt cag gca att 1104
Leu Cys Gly Glu Glu Gly Val Gly Lys Ala Leu Leu Ser Gln Ala Ile
355 360 365
cat aat gaa agc gag cgt gct gca ggt cct tat atc gcc gtc aat tgt 1152
His Asn Glu Ser Glu Arg Ala Ala Gly Pro Tyr Ile Ala Val Asn Cys
370 375 380
gag tta tat ggt gat gct gcg ctg gcg gaa gaa ttt att ggt ggc gat 1200
Glu Leu Tyr Gly Asp Ala Ala Leu Ala Glu Glu Phe Ile Gly Gly Asp
385 390 395 400
cgc acg gac aat gaa aat ggc cgt ctg agt cgg ctg gaa ctg gca cac 1248
Arg Thr Asp Asn Glu Asn Gly Arg Leu Ser Arg Leu Glu Leu Ala His
405 410 415
ggc ggc acg ctg ttt ctt gaa aag att gaa tat ctg gcg gtg gag tta 1296
Gly Gly Thr Leu Phe Leu Glu Lys Ile Glu Tyr Leu Ala Val Glu Leu
420 425 430
cag tct gct ttg ctt cag gtt atc aag cag ggg gtt atc acg cga ctg 1344
Gln Ser Ala Leu Leu Gln Val Ile Lys Gln Gly Val Ile Thr Arg Leu
435 440 445
gat gcg cgg cgt tta ata cca att gat gtc aaa gtg att gca aca acg 1392
Asp Ala Arg Arg Leu Ile Pro Ile Asp Val Lys Val Ile Ala Thr Thr
450 455 460
acc gcg gac ctc gca atg ctg gtg gaa caa aat cgt ttt agt cgc cag 1440
Thr Ala Asp Leu Ala Met Leu Val Glu Gln Asn Arg Phe Ser Arg Gln
465 470 475 480
ctg tat tac gcg ctg cat gca ttt gaa att acc atc ccg cct ctg cgt 1488
Leu Tyr Tyr Ala Leu His Ala Phe Glu Ile Thr Ile Pro Pro Leu Arg
485 490 495
atg cgg cgt ggc agc att ccg gcg ctg gtg aat aac aaa tta cgc agt 1536
Met Arg Arg Gly Ser Ile Pro Ala Leu Val Asn Asn Lys Leu Arg Ser
500 505 510
ctt gaa aaa cgc ttc tct acg cgg ctg aaa att gat gac gat gcc ctc 1584
Leu Glu Lys Arg Phe Ser Thr Arg Leu Lys Ile Asp Asp Asp Ala Leu
515 520 525
gct cgc ctg gtt tct tgt gca tgg cca ggc aac gat ttt gaa ctt tac 1632
Ala Arg Leu Val Ser Cys Ala Trp Pro Gly Asn Asp Phe Glu Leu Tyr
530 535 540
agc gtc atc gag aat ctt gct ctg agt agt gat aac ggg cgc att cgc 1680
Ser Val Ile Glu Asn Leu Ala Leu Ser Ser Asp Asn Gly Arg Ile Arg
545 550 555 560
gtc agt gat ttg ccg gaa cat ctg ttt acc gag cag gcg aca gat gat 1728
Val Ser Asp Leu Pro Glu His Leu Phe Thr Glu Gln Ala Thr Asp Asp
565 570 575
gtc agc gcc acc cgc ctt tcc acc agt ctg tca ttt gcg gaa gtt gaa 1776
Val Ser Ala Thr Arg Leu Ser Thr Ser Leu Ser Phe Ala Glu Val Glu
580 585 590
aaa gag gca att att aac gca gcc cag gtc aca ggc ggt cgc att cag 1824
Lys Glu Ala Ile Ile Asn Ala Ala Gln Val Thr Gly Gly Arg Ile Gln
595 600 605
gaa atg tcg gct tta ctt ggg atc ggc cgc act acg ctg tgg cgg aaa 1872
Glu Met Ser Ala Leu Leu Gly Ile Gly Arg Thr Thr Leu Trp Arg Lys
610 615 620
atg aag caa cat ggc att gat gca ggg cag ttt aag cgc cgg gta tga 1920
Met Lys Gln His Gly Ile Asp Ala Gly Gln Phe Lys Arg Arg Val
625 630 635
<210>67
<211>639
<212>PRT
<213>大肠杆菌
<400>67
Met Ser Gly Ala Phe Asn Asn Asp Gly Arg Gly Ile Ser Pro Leu Ile
1 5 10 15
Ala Thr Ser Trp Glu Arg Cys Asn Lys Leu Met Lys Arg Glu Thr Trp
20 25 30
Asn Val Pro His Gln Ala Gln Gly Val Thr Phe Ala Ser Ile Tyr Arg
35 40 45
Arg Lys Lys Ala Met Leu Thr Leu Gly Gln Ala Ala Leu Glu Asp Ala
50 55 60
Trp Glu Tyr Met Ala Pro Arg Glu Cys Ala Leu Phe Ile Leu Asp Glu
65 70 75 80
Thr Ala Cys Ile Leu Ser Arg Asn Gly Asp Pro Gln Thr Leu Gln Gln
85 90 95
Leu Ser Ala Leu Gly Phe Asn Asp Gly Thr Tyr Cys Ala Glu Gly Ile
100 105 110
Ile Gly Thr Cys Ala Leu Ser Leu Ala Ala Ile Ser Gly Gln Ala Val
115 120 125
Lys Thr Met Ala Asp Gln His Phe Lys Gln Val Leu Trp Asn Trp Ala
130 135 140
Phe Cys Ala Thr Pro Leu Phe Asp Ser Lys Gly Arg Leu Thr Gly Thr
145 150 155 160
Ile Ala Leu Ala Cys Pro Val Glu Gln Thr Thr Ala Ala Asp Leu Pro
165 170 175
Leu Thr Leu Ala Ile Ala Arg Glu Val Gly Asn Leu Leu Leu Thr Asp
180 185 190
Ser Leu Leu Ala Glu Thr Asn Arg His Leu Asn Gln Leu Asn Ala Leu
195 200 205
Leu Glu Ser Met Asp Asp Gly Val Ile Ser Trp Asp Glu Gln Gly Asn
210 215 220
Leu Gln Phe Ile Asn Ala Gln Ala Ala Arg Val Leu Arg Leu Asp Ala
225 230 235 240
Thr Ala Ser Gln Gly Arg Ala Ile Thr Glu Leu Leu Thr Leu Pro Ala
245 250 255
Val Leu Gln Gln Ala Ile Lys Gln Ala His Pro Leu Lys His Val Glu
260 265 270
Ala Thr Phe Glu Ser Gln His Gln Phe Ile Asp Ala Val Ile Thr Leu
275 280 285
Lys Pro Ile Ile Glu Thr Gln Gly Thr Ser Phe Ile Leu Leu Leu His
290 295 300
Pro Val Glu Gln Met Arg Gln Leu Met Thr Ser Gln Leu Gly Lys Val
305 310 315 320
Ser His Thr Phe Ala His Met Pro Gln Asp Asp Pro Gln Thr Arg Arg
325 330 335
Leu Ile His Phe Gly Arg Gln Ala Ala Arg Ser Ser Phe Pro Val Leu
340 345 350
Leu Cys Gly Glu Glu Gly Val Gly Lys Ala Leu Leu Ser Gln Ala Ile
355 360 365
His Asn Glu Ser Glu Arg Ala Ala Gly Pro Tyr Ile Ala Val Asn Cys
370 375 380
Glu Leu Tyr Gly Asp Ala Ala Leu Ala Glu Glu Phe Ile Gly Gly Asp
385 390 395 400
Arg Thr Asp Asn Glu Asn Gly Arg Leu Ser Arg Leu Glu Leu Ala His
405 410 415
Gly Gly Thr Leu Phe Leu Glu Lys Ile Glu Tyr Leu Ala Val Glu Leu
420 425 430
Gln Ser Ala Leu Leu Gln Val Ile Lys Gln Gly Val Ile Thr Arg Leu
435 440 445
Asp Ala Arg Arg Leu Ile Pro Ile Asp Val Lys Val Ile Ala Thr Thr
450 455 460
Thr Ala Asp Leu Ala Met Leu Val Glu Gln Asn Arg Phe Ser Arg Gln
465 470 475 480
Leu Tyr Tyr Ala Leu His Ala Phe Glu Ile Thr Ile Pro Pro Leu Arg
485 490 495
Met Arg Arg Gly Ser Ile Pro Ala Leu Val Asn Asn Lys Leu Arg Ser
500 505 510
Leu Glu Lys Arg Phe Ser Thr Arg Leu Lys Ile Asp Asp Asp Ala Leu
515 520 525
Ala Arg Leu Val Ser Cys Ala Trp Pro Gly Asn Asp Phe Glu Leu Tyr
530 535 540
Ser Val Ile Glu Asn Leu Ala Leu Ser Ser Asp Asn Gly Arg Ile Arg
545 550 555 560
Val Ser Asp Leu Pro Glu His Leu Phe Thr Glu Gln Ala Thr Asp Asp
565 570 575
Val Ser Ala Thr Arg Leu Ser Thr Ser Leu Ser Phe Ala Glu Val Glu
580 585 590
Lys Glu Ala Ile Ile Asn Ala Ala Gln Val Thr Gly Gly Arg Ile Gln
595 600 605
Glu Met Ser Ala Leu Leu Gly Ile Gly Arg Thr Thr Leu Trp Arg Lys
610 615 620
Met Lys Gln His Gly Ile Asp Ala Gly Gln Phe Lys Arg Arg Val
625 630 635
<210>68
<211>663
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(663)
<223>
<400>68
atg gaa ctg tat ctg gat act tca gac gtt gtt gcg gtg aag gcg ctg 48
Met Glu Leu Tyr Leu Asp Thr Ser Asp Val Val Ala Val Lys Ala Leu
1 5 10 15
tca cgt att ttt ccg ctg gcg ggt gtg acc act aac cca agc att atc 96
Ser Arg Ile Phe Pro Leu Ala Gly Val Thr Thr Asn Pro Ser Ile Ile
20 25 30
gcc gcg ggt aaa aaa ccg ctg gat gtt gtg ctt ccg caa ctt cat gaa 144
Ala Ala Gly Lys Lys Pro Leu Asp Val Val Leu Pro Gln Leu His Glu
35 40 45
gcg atg ggc ggt cag ggg cgt ctg ttt gcc cag gta atg gct acc act 192
Ala Met Gly Gly Gln Gly Arg Leu Phe Ala Gln Val Met Ala Thr Thr
50 55 60
gcc gaa ggg atg gtt aat gac gcg ctt aag ctg cgt tct att att gcg 240
Ala Glu Gly Met Val Asn Asp Ala Leu Lys Leu Arg Ser Ile Ile Ala
65 70 75 80
gat atc gtg gtg aaa gtt ccg gtg acc gcc gag ggg ctg gca gct att 288
Asp Ile Val Val Lys Val Pro Val Thr Ala Glu Gly Leu Ala Ala Ile
85 90 95
aag atg tta aaa gcg gaa ggg att ccg acg ctg gga acc gcg gta tat 336
Lys Met Leu Lys Ala Glu Gly Ile Pro Thr Leu Gly Thr Ala Val Tyr
100 105 110
ggc gca gca caa ggg ctg ctg tcg gcg ctg gca ggt gcg gaa tat gtt 384
Gly Ala Ala Gln Gly Leu Leu Ser Ala Leu Ala Gly Ala Glu Tyr Val
115 120 125
gcg cct tac gtt aat cgt att gat gct cag ggc ggt agc ggc att cag 432
Ala Pro Tyr Val Asn Arg Ile Asp Ala Gln Gly Gly Ser Gly Ile Gln
130 135 140
act gtg acc gac tta cac cag tta ttg aaa atg cat gcg ccg cag gcg 480
Thr Val Thr Asp Leu His Gln Leu Leu Lys Met His Ala Pro Gln Ala
145 150 155 160
aaa gtg ctg gca gcg agt ttc aaa acc ccg cgt cag gcg ctg gac tgc 528
Lys Val Leu Ala Ala Ser Phe Lys Thr Pro Arg Gln Ala Leu Asp Cys
165 170 175
tta ctg gca gga tgt gaa tca att act ctg cca ctg gat gtg gca caa 576
Leu Leu Ala Gly Cys Glu Ser Ile Thr Leu Pro Leu Asp Val Ala Gln
180 185 190
cag atg att agc tat ccg gcg gtt gat gcc gct gtg gcg aag ttt gag 624
Gln Met Ile Ser Tyr Pro Ala Val Asp Ala Ala Val Ala Lys Phe Glu
195 200 205
cag gac tgg cag gga gcg ttt ggc aga acg tcg att taa 663
Gln Asp Trp Gln Gly Ala Phe Gly Arg Thr Ser Ile
210 215 220
<210>69
<211>220
<212>PRT
<213>大肠杆菌
<400>69
Met Glu Leu Tyr Leu Asp Thr Ser Asp Val Val Ala Val Lys Ala Leu
1 5 10 15
Ser Arg Ile Phe Pro Leu Ala Gly Val Thr Thr Asn Pro Ser Ile Ile
20 25 30
Ala Ala Gly Lys Lys Pro Leu Asp Val Val Leu Pro Gln Leu His Glu
35 40 45
Ala Met Gly Gly Gln Gly Arg Leu Phe Ala Gln Val Met Ala Thr Thr
50 55 60
Ala Glu Gly Met Val Asn Asp Ala Leu Lys Leu Arg Ser Ile Ile Ala
65 70 75 80
Asp Ile Val Val Lys Val Pro Val Thr Ala Glu Gly Leu Ala Ala Ile
85 90 95
Lys Met Leu Lys Ala Glu Gly Ile Pro Thr Leu Gly Thr Ala Val Tyr
100 105 110
Gly Ala Ala Gln Gly Leu Leu Ser Ala Leu Ala Gly Ala Glu Tyr Val
115 120 125
Ala Pro Tyr Val Asn Arg Ile Asp Ala Gln Gly Gly Ser Gly Ile Gln
130 135 140
Thr Val Thr Asp Leu His Gln Leu Leu Lys Met His Ala Pro Gln Ala
145 150 155 160
Lys Val Leu Ala Ala Ser Phe Lys Thr Pro Arg Gln Ala Leu Asp Cys
165 170 175
Leu Leu Ala Gly Cys Glu Ser Ile Thr Leu Pro Leu Asp Val Ala Gln
180 185 190
Gln Met Ile Ser Tyr Pro Ala Val Asp Ala Ala Val Ala Lys Phe Glu
195 200 205
Gln Asp Trp Gln Gly Ala Phe Gly Arg Thr Ser Ile
210 215 220
<210>70
<211>663
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(663)
<223>
<400>70
atg gaa ctg tat ctg gac acc gct aac gtc gca gaa gtc gaa cgt ctg 48
Met Glu Leu Tyr Leu Asp Thr Ala Asn Val Ala Glu Val Glu Arg Leu
1 5 10 15
gca cgc ata ttc ccc att gcc ggg gtg aca act aac ccg agc att atc 96
Ala Arg Ile Phe Pro Ile Ala Gly Val Thr Thr Asn Pro Ser Ile Ile
20 25 30
gct gcc agc aag gag tcc ata tgg gaa gtg ctg ccg cgt ctg caa aaa 144
Ala Ala Ser Lys Glu Ser Ile Trp Glu Val Leu Pro Arg Leu Gln Lys
35 40 45
gcg att ggt gat gag ggc att ctg ttt gct cag acc atg agc cgc gac 192
Ala Ile Gly Asp Glu Gly Ile Leu Phe Ala Gln Thr Met Ser Arg Asp
50 55 60
gcg cag ggg atg gtg gaa gaa gcg aag cgc ctg cgc gac gct att ccg 240
Ala Gln Gly Met Val Glu Glu Ala Lys Arg Leu Arg Asp Ala Ile Pro
65 70 75 80
ggt att gtg gtg aaa atc ccg gtg act tcc gaa ggt ctg gca gca att 288
Gly Ile Val Val Lys Ile Pro Val Thr Ser Glu Gly Leu Ala Ala Ile
85 90 95
aaa ata ctg aaa aaa gag ggt att act aca ctt ggc act gct gta tat 336
Lys Ile Leu Lys Lys Glu Gly Ile Thr Thr Leu Gly Thr Ala Val Tyr
100 105 110
agc gcc gca caa ggg tta tta gcc gca ctg gca ggg gca aaa tac gtt 384
Ser Ala Ala Gln Gly Leu Leu Ala Ala Leu Ala Gly Ala Lys Tyr Val
115 120 125
gct ccg tat gtt aac cgc gta gat gcc cag ggc gga gac ggc att cgt 432
Ala Pro Tyr Val Asn Arg Val Asp Ala Gln Gly Gly Asp Gly Ile Arg
130 135 140
acg gtt cag gag ctg caa acg ctg tta gaa atg cac gcg cca gaa agc 480
Thr Val Gln Glu Leu Gln Thr Leu Leu Glu Met His Ala Pro Glu Ser
145 150 155 160
atg gtg ctg gca gcc agc ttt aaa acg ccg cgt cag gcg ctg gac tgt 528
Met Val Leu Ala Ala Ser Phe Lys Thr Pro Arg Gln Ala Leu Asp Cys
165 170 175
tta ctg gca gga tgt gaa tcc atc acc ctg ccc tta gat gta gcg caa 576
Leu Leu Ala Gly Cys Glu Ser Ile Thr Leu Pro Leu Asp Val Ala Gln
180 185 190
caa atg ctc aac acc cct gcg gta gag tca gct ata gag aag ttc gaa 624
Gln Met Leu Asn Thr Pro Ala Val Glu Ser Ala Ile Glu Lys Phe Glu
195 200 205
cac gac tgg aat gcc gca ttt ggc act act cat ctc taa 663
His Asp Trp Asn Ala Ala Phe Gly Thr Thr His Leu
210 215 220
<210>71
<211>220
<212>PRT
<213>大肠杆菌
<400>71
Met Glu Leu Tyr Leu Asp Thr Ala Asn Val Ala Glu Val Glu Arg Leu
1 5 10 15
Ala Arg Ile Phe Pro Ile Ala Gly Val Thr Thr Asn Pro Ser Ile Ile
20 25 30
Ala Ala Ser Lys Glu Ser Ile Trp Glu Val Leu Pro Arg Leu Gln Lys
35 40 45
Ala Ile Gly Asp Glu Gly Ile Leu Phe Ala Gln Thr Met Ser Arg Asp
50 55 60
Ala Gln Gly Met Val Glu Glu Ala Lys Arg Leu Arg Asp Ala Ile Pro
65 70 75 80
Gly Ile Val Val Lys Ile Pro Val Thr Ser Glu Gly Leu Ala Ala Ile
85 90 95
Lys Ile Leu Lys Lys Glu Gly Ile Thr Thr Leu Gly Thr Ala Val Tyr
100 105 110
Ser Ala Ala Gln Gly Leu Leu Ala Ala Leu Ala Gly Ala Lys Tyr Val
115 120 125
Ala Pro Tyr Val Asn Arg Val Asp Ala Gln Gly Gly Asp Gly Ile Arg
130 135 140
Thr Val Gln Glu Leu Gln Thr Leu Leu Glu Met His Ala Pro Glu Ser
145 150 155 160
Met Val Leu Ala Ala Ser Phe Lys Thr Pro Arg Gln Ala Leu Asp Cys
165 170 175
Leu Leu Ala Gly Cys Glu Ser Ile Thr Leu Pro Leu Asp Val Ala Gln
180 185 190
Gln Met Leu Asn Thr Pro Ala Val Glu Ser Ala Ile Glu Lys Phe Glu
195 200 205
His Asp Trp Asn Ala Ala Phe Gly Thr Thr His Leu
210 215 220
<210>72
<211>1053
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1053)
<223>
<400>72
atg aca gat att gcg cag ttg ctt ggc aaa gac gcc gac aac ctt tta 48
Met Thr Asp Ile Ala Gln Leu Leu Gly Lys Asp Ala Asp Asn Leu Leu
1 5 10 15
cag cac cgt tgt atg aca att cct tct gac cag ctt tat ctc ccc gga 96
Gln His Arg Cys Met Thr Ile Pro Ser Asp Gln Leu Tyr Leu Pro Gly
20 25 30
cat gac tac gta gac cgc gta atg att gac aat aat cgc ccg cca gcg 144
His Asp Tyr Val Asp Arg Val Met Ile Asp Asn Asn Arg Pro Pro Ala
35 40 45
gtg tta cgt aat atg cag acg ttg tac aac acc ggg cgt ctg gct ggc 192
Val Leu Arg Asn Met Gln Thr Leu Tyr Asn Thr Gly Arg Leu Ala Gly
50 55 60
aca gga tat ctt tct att ctg ccg gtt gac cag ggc gtt gag cac tct 240
Thr Gly Tyr Leu Ser Ile Leu Pro Val Asp Gln Gly Val Glu His Ser
65 70 75 80
gcc gga gct tca ttt gct gct aac ccg ctc tac ttt gac ccg aaa aac 288
Ala Gly Ala Ser Phe Ala Ala Asn Pro Leu Tyr Phe Asp Pro Lys Asn
85 90 95
att gtt gaa ctg gcg atc gaa gcg ggc tgt aac tgt gtg gcg tca act 336
Ile Val Glu Leu Ala Ile Glu Ala Gly Cys Asn Cys Val Ala Ser Thr
100 105 110
tac ggc gtg ctg gcg tcg gta tcg cgg cgt tat gcg cat cgc att cca 384
Tyr Gly Val Leu Ala Ser Val Ser Arg Arg Tyr Ala His Arg Ile Pro
115 120 125
ttc ctc gtc aaa ctt aat cac aac gag acg cta agt tac ccg aat acc 432
Phe Leu Val Lys Leu Asn His Asn Glu Thr Leu Ser Tyr Pro Asn Thr
130 135 140
tac gat caa acg ctg tat gcc agc gtg gag cag gcg ttc aac atg ggc 480
Tyr Asp Gln Thr Leu Tyr Ala Ser Val Glu Gln Ala Phe Asn Met Gly
145 150 155 160
gcg gtt gcg gtt ggt gcg act atc tat ttt ggc tcg gaa gag tca cgt 528
Ala Val Ala Val Gly Ala Thr Ile Tyr Phe Gly Ser Glu Glu Ser Arg
165 170 175
cgc cag att gaa gaa att tct gcg gct ttt gaa cgt gcg cac gag ctg 576
Arg Gln Ile Glu Glu Ile Ser Ala Ala Phe Glu Arg Ala His Glu Leu
180 185 190
ggt atg gtg aca gtg ctg tgg gcc tat ttg cgt aac tcc gcc ttt aag 624
Gly Met Val Thr Val Leu Trp Ala Tyr Leu Arg Ash Ser Ala Phe Lys
195 200 205
aaa gat ggc gtt gat tac cat gtt tcc gcc gac ctg acc ggt cag gca 672
Lys Asp Gly Val Asp Tyr His Val Ser Ala Asp Leu Thr Gly Gln Ala
210 215 220
aac cat ctg gcg gca acc atc ggt gca gat atc gtc aaa caa aaa atg 720
Asn His Leu Ala Ala Thr Ile Gly Ala Asp Ile Val Lys Gln Lys Met
225 230 235 240
gcg gaa aat aac ggc ggc tat aaa gca att aat tac ggt tac acc gac 768
Ala Glu Asn Asn Gly Gly Tyr Lys Ala Ile Asn Tyr Gly Tyr Thr Asp
245 250 255
gat cgt gtt tac agc aaa ttg acc agc gaa aac ccg att gat ctg gtg 816
Asp Arg Val Tyr Ser Lys Leu Thr Ser Glu Asn Pro Ile Asp Leu Val
260 265 270
cgt tat cag tta gct aac tgc tat atg ggt cgg gct ggg ttg ata aac 864
Arg Tyr Gln Leu Ala Asn Cys Tyr Met Gly Arg Ala Gly Leu Ile Asn
275 280 285
tcc ggc ggt gct gcg ggc ggt gaa act gac ctc agc gat gca gtg cgt 912
Ser Gly Gly Ala Ala Gly Gly Glu Thr Asp Leu Ser Asp Ala Val Arg
290 295 300
act gcg gtt atc aac aaa cgc gca ggc gga atg ggg ctg att ctt gga 960
Thr Ala Val Ile Asn Lys Arg Ala Gly Gly Met Gly Leu Ile Leu Gly
305 310 315 320
cgt aaa gcg ttc aag aaa tcg atg gct gac ggc gtg aaa ctg att aac 1008
Arg Lys Ala Phe Lys Lys Ser Met Ala Asp Gly Val Lys Leu Ile Asn
325 330 335
gcc gtg cag gac gtt tat ctc gat agc aaa att act atc gcc tga 1053
Ala Val Gln Asp Val Tyr Leu Asp Ser Lys Ile Thr Ile Ala
340 345 350
<210>73
<211>350
<212>PRT
<213>大肠杆菌
<400>73
Met Thr Asp Ile Ala Gln Leu Leu Gly Lys Asp Ala Asp Asn Leu Leu
1 5 10 15
Gln His Arg Cys Met Thr Ile Pro Ser Asp Gln Leu Tyr Leu Pro Gly
20 25 30
His Asp Tyr Val Asp Arg Val Met Ile Asp Asn Asn Arg Pro Pro Ala
35 40 45
Val Leu Arg Asn Met Gln Thr Leu Tyr Asn Thr Gly Arg Leu Ala Gly
50 55 60
Thr Gly Tyr Leu Ser Ile Leu Pro Val Asp Gln Gly Val Glu His Ser
65 70 75 80
Ala Gly Ala Ser Phe Ala Ala Asn Pro Leu Tyr Phe Asp Pro Lys Asn
85 90 95
Ile Val Glu Leu Ala Ile Glu Ala Gly Cys Asn Cys Val Ala Ser Thr
100 105 110
Tyr Gly Val Leu Ala Ser Val Ser Arg Arg Tyr Ala His Arg Ile Pro
115 120 125
Phe Leu Val Lys Leu Asn His Asn Glu Thr Leu Ser Tyr Pro Asn Thr
130 135 140
Tyr Asp Gln Thr Leu Tyr Ala Ser Val Glu Gln Ala Phe Asn Met Gly
145 150 155 160
Ala Val Ala Val Gly Ala Thr Ile Tyr Phe Gly Ser Glu Glu Ser Arg
165 170 175
Arg Gln Ile Glu Glu Ile Ser Ala Ala Phe Glu Arg Ala His Glu Leu
180 185 190
Gly Met Val Thr Val Leu Trp Ala Tyr Leu Arg Asn Ser Ala Phe Lys
195 200 205
Lys Asp Gly Val Asp Tyr His Val Ser Ala Asp Leu Thr Gly Gln Ala
210 215 220
Asn His Leu Ala Ala Thr Ile Gly Ala Asp Ile Val Lys Gln Lys Met
225 230 235 240
Ala Glu Asn Asn Gly Gly Tyr Lys Ala Ile Asn Tyr Gly Tyr Thr Asp
245 250 255
Asp Arg Val Tyr Ser Lys Leu Thr Ser Glu Asn Pro Ile Asp Leu Val
260 265 270
Arg Tyr Gln Leu Ala Asn Cys Tyr Met Gly Arg Ala Gly Leu Ile Asn
275 280 285
Ser Gly Gly Ala Ala Gly Gly Glu Thr Asp Leu Ser Asp Ala Val Arg
290 295 300
Thr Ala Val Ile Asn Lys Arg Ala Gly Gly Met Gly Leu Ile Leu Gly
305 310 315 320
Arg Lys Ala Phe Lys Lys Ser Met Ala Asp Gly Val Lys Leu Ile Asn
325 330 335
Ala Val Gln Asp Val Tyr Leu Asp Ser Lys Ile Thr Ile Ala
340 345 350
<210>74
<211>1143
<212>DNA
<213>痢疾志贺氏菌(Shigella dysenteriae)
<220>
<221>CDS
<222>(1)..(1143)
<223>
<400>74
atg ccg cat ttg gca cta ctc atc tct aaa gga gca att atg gac cgc 48
Met Pro His Leu Ala Leu Leu Ile Ser Lys Gly Ala Ile Met Asp Arg
1 5 10 15
att att caa tca ccg ggt aaa tac atc cag ggc gct gat gtg att aat 96
Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp Val Ile Asn
20 25 30
cgt ctg ggc gaa tac ctg aag ccg ctg gca gaa ctc tgg tta gtg gtg 144
Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Leu Trp Leu Val Val
35 40 45
ggt gac aaa ttt gtt tta ggt ttt gct caa tcc act gtc gag aaa agc 192
Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val Glu Lys Ser
50 55 60
ttt aaa gat gct gga ctg gta gta gaa att gcg ccg ttt ggc ggt gaa 240
Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe Gly Gly Glu
65 70 75 80
tgt tcg caa aat gag atc gac cgt ctg cgt ggc atc gcg gag act gcg 288
Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala Glu Thr Ala
85 90 95
cag tgt ggc gca att ctc ggt atc ggt ggc gga aaa act ttc gat act 336
Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr Phe Asp Thr
100 105 110
gcc aaa gca ctg gca cat ttc atg ggt gtt ccg gta gcg atc gca ccg 384
Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala Ile Ala Pro
115 120 125
acg atc gcc tct acc gac gca ccg tgc agc gca ttg tct gtt atc tac 432
Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser Val Ile Tyr
130 135 140
acc gat gag ggt gag ttt gac cgc tat ctg ctg ttg cca aat aac cct 480
Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro Asn Asn Pro
145 150 155 160
aat atg gtc att gtc gac acc aaa atc gtc gct ggc gca cct gca cgt 528
Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Ala Pro Ala Arg
165 170 175
ctg tta gcg gcg ggt atc ggc gat gcg ctg gca acc tgg ttt gaa gcg 576
Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp Phe Glu Ala
180 185 190
cgt gcc tgc tct cgt agc ggc gcg acc acc atg gcg ggc ggc aag tgc 624
Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly Gly Lys Cys
195 200 205
acc cag gct gcg ctg gca ctg gct gaa ctg tgc tac aac acc ctg ctg 672
Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn Thr Leu Leu
210 215 220
gaa gaa ggc gaa aaa gcg atg ctt gct gcc gaa cag cat gta gtg act 720
Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His Val Val Thr
225 230 235 240
ccg gcg ctg gag cgc gtg att gaa gcg aac acc tat ttg agc ggt gtt 768
Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu Ser Gly Val
245 250 255
ggt ttt gaa agt ggt ggt ctg gct gcg gcg cac gca gtg cat aac ggc 816
Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val His Asn Gly
260 265 270
ctg acc gct atc ccg gac gcg cat cac tat tat cac ggt gaa aaa gtg 864
Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly Glu Lys Val
275 280 285
gca ttc ggt acg ctg acg cag ctg gtt ctg gaa aat gcg ccg gtg gag 912
Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala Pro Val Glu
290 295 300
gaa atc gaa acc gta gct gcc ctt agc cat gcg gta ggt ttg cca ata 960
Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly Leu Pro Ile
305 310 315 320
act ctc gct caa ctg gat att aaa gaa gat gtc ccg gcg aaa atg cga 1008
Thr Leu Ala Gin Leu Asp Ile Lys Glu Asp Val Pro Ala Lys Met Arg
325 330 335
att gtg gca gaa gcg gca tgt gca gaa ggt gaa acc att cac aac atg 1056
Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile His Asn Met
340 345 350
cct ggc ggc gcg acg cca gat cag gtt tac gcc gca ctg ctg gta gct 1104
Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu Leu Val Ala
355 360 365
gac cag tat gga caa cgt ttc ctg caa gag tgg gaa taa 1143
Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu
370 375 380
<210>75
<211>380
<212>PRT
<213>痢疾志贺氏菌
<400>75
Met Pro His Leu Ala Leu Leu Ile Ser Lys Gly Ala Ile Met Asp Arg
1 5 10 15
Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp Val Ile Asn
20 25 30
Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Leu Trp Leu Val Val
35 40 45
Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val Glu Lys Ser
50 55 60
Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe Gly Gly Glu
65 70 75 80
Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala Glu Thr Ala
85 90 95
Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr Phe Asp Thr
100 105 110
Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala Ile Ala Pro
115 120 125
Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser Val Ile Tyr
130 135 140
Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro Asn Asn Pro
145 150 155 160
Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Ala Pro Ala Arg
165 170 175
Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp Phe Glu Ala
180 185 190
Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly Gly Lys Cys
195 200 205
Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn Thr Leu Leu
210 215 220
Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His Val Val Thr
225 230 235 240
Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu Ser Gly Val
245 250 255
Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val His Asn Gly
260 265 270
Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly Glu Lys Val
275 280 285
Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala Pro Val Glu
290 295 300
Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly Leu Pro Ile
305 310 315 320
Thr Leu Ala Gln Leu Asp Ile Lys Glu Asp Val Pro Ala Lys Met Arg
325 330 335
Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile His Asn Met
340 345 350
Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu Leu Val Ala
355 360 365
Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu
370 375 380
<210>76
<211>1104
<212>DNA
<213>伤寒沙门氏菌(Salmonella typhimurium)
<220>
<22l>CDS
<222>(1)..(1104)
<223>
<400>76
atg gat cgc att att cag tca cca ggt aag tat att cag ggt gca aac 48
Met Asp Arg Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asn
1 5 10 15
gtc atc gcg cgt ctt ggc gat tat tta aaa cca atg gcg aac aac tgg 96
Val Ile Ala Arg Leu Gly Asp Tyr Leu Lys Pro Met Ala Asn Asn Trp
20 25 30
ctg gtt gtg ggc gat aaa ttc gtg ctg gga ttt gcc gaa gag acg ctg 144
Leu Val Val Gly Asp Lys Phe Val Leu Gly Phe Ala Glu Glu Thr Leu
35 40 45
cgc aaa agc ctg acg gat gcc ggt ttg tca gta gaa atc gcc ccg ttt 192
Arg Lys Ser Leu Thr Asp Ala Gly Leu Ser Val Glu Ile Ala Pro Phe
50 55 60
ggc ggc gaa tgt tcg caa aat gag atc gac agg ctg cgc gcc gtc gcc 240
Gly Gly Glu Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Ala Val Ala
65 70 75 80
gaa aaa agt cag tgt ggc gcc gta ctg ggt atc ggc ggc ggt aaa acg 288
Glu Lys Ser Gln Cys Gly Ala Val Leu Gly Ile Gly Gly Gly Lys Thr
85 90 95
ctg gat acc gcc aaa gcg ctg gcg cac ttt atg aac gtc ccg gtc gct 336
Leu Asp Thr Ala Lys Ala Leu Ala His Phe Met Asn Val Pro Val Ala
100 105 110
atc gcg ccg acc atc gcc tct acc gac gca ccg tgc agc gca ctc tcg 384
Ile Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser
115 120 125
gtt att tat acc gat gcc ggt gag ttt gac cgt tat ctg ctg ctg ccg 432
Val Ile Tyr Thr Asp Ala Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro
130 135 140
cat aac ccg aat atg gtt att gtc gat acg cag ata gtg gcg ggc gcg 480
His Asn Pro Asn Met Val Ile Val Asp Thr Gln Ile Val Ala Gly Ala
145 150 155 160
ccg gcg cgt ctg ctg gca gcc ggt atc ggc gat gca ctg gcg acc tgg 528
Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp
165 170 175
ttt gaa gcg cgc gcc tgc tca cgc agc ggc gcc acc aca atg gcg ggc 576
Phe Glu Ala Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly
180 185 190
ggc aag tgt aca cag gcc gcg ctg gcg ctg gcg gag cta tgc tat aac 624
Gly Lys Cys Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn
195 200 205
acg ctg atc gaa gaa ggc gaa aaa gcc atg ttg gcc gcc gaa cag cac 672
Thr Leu Ile Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His
210 215 220
gtc gtc acg cca gcg ctg gaa cgc gtc atc gaa gcc aac acc tac ctg 720
Val Val Thr Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu
225 230 235 240
agc ggg gtc ggt ttt gaa agc ggc ggt ctg gcc gca gcg cac gcg att 768
Ser Gly Val Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Ile
245 250 255
cat aac ggt tta acg gcg att ccg gat gcg cac cac tat tat cac ggt 816
His Asn Gly Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly
260 265 270
gag aag gtc gct ttc ggt acg ctg acg caa ctg gtg ctg gaa aac gcg 864
Glu Lys Val Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala
275 280 285
ccg gtc gaa gaa atc gaa acc gtt gcg gcg ctg tgc cat tcc gtt ggc 912
Pro Val Glu Glu Ile Glu Thr Val Ala Ala Leu Cys His Ser Val Gly
290 295 300
ctg ccg att acg ctg gcg caa ctg gat att aaa cag gat att ccg gcc 960
Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Lys Gln Asp Ile Pro Ala
305 310 315 320
aag atg cgc acc gtc gcg gaa gcc tcc tgc gca gaa ggt gaa act att 1008
Lys Met Arg Thr Val Ala Glu Ala Ser Cys Ala Glu Gly Glu Thr Ile
325 330 335
cat aac atg cct ggc ggc gca acg ccg gat gaa gtg tac gcc gcg ctg 1056
His Asn Met Pro Gly Gly Ala Thr Pro Asp Glu Val Tyr Ala Ala Leu
340 345 350
ctg gtc gcc gac cag tac ggt caa cgc ttc ttg cag gaa tgg gaa taa 1104
Leu Val Ala Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu
355 360 365
<210>77
<211>367
<212>PRT
<213>伤寒沙门氏菌
<400>77
Met Asp Arg Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asn
1 5 10 15
Val Ile Ala Arg Leu Gly Asp Tyr Leu Lys Pro Met Ala Asn Asn Trp
20 25 30
Leu Val Val Gly Asp Lys Phe Val Leu Gly Phe Ala Glu Glu Thr Leu
35 40 45
Arg Lys Ser Leu Thr Asp Ala Gly Leu Ser Val Glu Ile Ala Pro Phe
50 55 60
Gly Gly Glu Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Ala Val Ala
65 70 75 80
Glu Lys Ser Gln Cys Gly Ala Val Leu Gly Ile Gly Gly Gly Lys Thr
85 90 95
Leu Asp Thr Ala Lys Ala Leu Ala His Phe Met Asn Val Pro Val Ala
100 105 110
Ile Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser
115 120 125
Val Ile Tyr Thr Asp Ala Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro
130 135 140
His Asn Pro Asn Met Val Ile Val Asp Thr Gln Ile Val Ala Gly Ala
145 150 155 160
Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp
165 170 175
Phe Glu Ala Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly
180 185 190
Gly Lys Cys Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn
195 200 205
Thr Leu Ile Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His
210 215 220
Val Val Thr Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu
225 230 235 240
Ser Gly Val Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Ile
245 250 255
His Asn Gly Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly
260 265 270
Glu Lys Val Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala
275 280 285
Pro Val Glu Glu Ile Glu Thr Val Ala Ala Leu Cys His Ser Val Gly
290 295 300
Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Lys Gln Asp Ile Pro Ala
305 310 315 320
Lys Met Arg Thr Val Ala Glu Ala Ser Cys Ala Glu Gly Glu Thr Ile
325 330 335
His Asn Met Pro Gly Gly Ala Thr Pro Asp Glu Val Tyr Ala Ala Leu
340 345 350
Leu Val Ala Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu
355 360 365
<210>78
<211>1098
<212>DNA
<213>恶臭假单胞菌
<220>
<221>CDS
<222>(1)..(1098)
<223>
<400>78
atg gac cgc gcc att caa tca ccc ggc aaa tat gta caa ggg gcc gat 48
Met Asp Arg Ala Ile Gln Ser Pro Gly Lys Tyr Val Gln Gly Ala Asp
1 5 10 15
gcg ctg caa cgg ctg ggg gac tac ctc aag ccg ctg gcg gac agc tgg 96
Ala Leu Gln Arg Leu Gly Asp Tyr Leu Lys Pro Leu Ala Asp Ser Trp
20 25 30
ctg gtg att gcc gac aag ttc gtg ctg ggc ttt gcc gaa gac acc atc 144
Leu Val Ile Ala Asp Lys Phe Val Leu Gly Phe Ala Glu Asp Thr Ile
35 40 45
cgc caa agc ctc agc aag gcc ggg ctg gcc atg gac atc gtc gcc ttc 192
Arg Gln Ser Leu Ser Lys Ala Gly Leu Ala Met Asp Ile Val Ala Phe
50 55 60
aac ggc gaa tgc tcg cag ggc gag gtc gat cgc ctg tgc caa ctg gcc 240
Asn Gly Glu Cys Ser Gln Gly Glu Val Asp Arg Leu Cys Gln Leu Ala
65 70 75 80
acg caa aac ggg cgc agc gcc atc gtc ggc att ggt ggc ggc aag acg 288
Thr Gln Asn Gly Arg Ser Ala Ile Val Gly Ile Gly Gly Gly Lys Thr
85 90 95
ctg gac acc gcc aag gcc gtg gcc ttt ttc cag aaa gtg ccc gtg gcc 336
Leu Asp Thr Ala Lys Ala Val Ala Phe Phe Gln Lys Val Pro Val Ala
100 105 110
gtg gcc ccc acc atc gcc tcc acc gac gcg ccc tgc agc gcg ctg tcg 384
Val Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser
115120125
gtg ctc tat acc gat gaa ggt gag ttc gac cgc tat ctg atg ctg ccc 432
Val Leu Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Met Leu Pro
130 135 140
acc aac ccc gcc ctg gtg gtg gtg gac acc gcc atc gtc gcc cgt gca 480
Thr Asn Pro Ala Leu Val Val Val Asp Thr Ala Ile Val Ala Arg Ala
145 150 155 160
ccg gcg cgg ctg ttg gcg gcc ggc att ggt gat gcc ctg gcc acc tgg 528
Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp
165 170 175
ttc gag gcg cgt gcc gca tcg cgc agc agc gct gcc acc atg gcc ggc 576
Phe Glu Ala Arg Ala Ala Ser Arg Ser Ser Ala Ala Thr Met Ala Gly
180 185 190
ggc ccg gcc acg cag acc gca ctg aac ctg gcc agg ttc tgc tac gac 624
Gly Pro Ala Thr Gln Thr Ala Leu Asn Leu Ala Arg Phe Cys Tyr Asp
195 200 205
acc ctg ctg gaa gag ggt gaa aaa gcc atg ttg gcc gtg cag gcc cag 672
Thr Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Val Gln Ala Gln
210 215 220
gtg gtg acg ccg gcg ctg gag cgc atc gtc gag gcc aac acc tat ctg 720
Val Val Thr Pro Ala Leu Glu Arg Ile ValGlu Ala Asn Thr Tyr Leu
225 230 235 240
agc ggg gtc ggg ttt gaa agc ggt ggc gtg gcc gcc gcc cac gcg gtg 768
Ser Gly Val Gly Phe Glu Ser Gly Gly Val Ala Ala Ala His Ala Val
245 250 255
cac aac ggc ctg acc gcc gtg gcc gaa acc cac cac ttc tac cac ggc 816
His Asn Gly Leu Thr Ala Val Ala Glu Thr His His Phe Tyr His Gly
260 265 270
gaa aaa gtg gcg ttt ggc gtg ctg gtg caa ctg gcg ctg gaa aac gcc 864
Glu Lys Val Ala Phe Gly Val Leu Val Gln Leu Ala Leu Glu Asn Ala
275 280 285
tcc aac gcg gaa atg cag gaa gtg atg tcg ctg tgc cac gcc gtg ggc 912
Ser Asn Ala Glu Met Gln Glu Val Met Ser Leu Cys His Ala Val Gly
290 295 300
ctg ccc atc acg ctg gcg cag ctg gac att acc gaa gac atc ccc acc 960
Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Thr Glu Asp Ile Pro Thr
305 310 315 320
aag atg cgc gcc gtg gcc gag ctg gcc tgc gcc cca ggc gag acc atc 1008
Lys Met Arg Ala Val Ala Glu Leu Ala Cys Ala Pro Gly Glu Thr Ile
325 330 335
cac aac atg ccc ggc ggc gtg acg gtg gag cag gtc tat ggc gcg ctg 1056
His Asn Met Pro Gly Gly Val Thr Val Glu Gln Val Tyr Gly Ala Leu
340 345 350
ctg gtg gcg gac cag ctg ggg cag cat ttt ctg gag ttt tga 1098
Leu Val Ala Asp Gln Leu Gly Gln His Phe Leu Glu Phe
355 360 365
<210>79
<211>365
<212>PRT
<213>恶臭假单胞菌(Pseudomonas putida)
<400>79
Met Asp Arg Ala Ile Gln Ser Pro Gly Lys Tyr Val Gln Gly Ala Asp
1 5 10 15
Ala Leu Gln Arg Leu Gly Asp Tyr Leu Lys Pro Leu Ala Asp Ser Trp
20 25 30
Leu Val Ile Ala Asp Lys Phe Val Leu Gly Phe Ala Glu Asp Thr Ile
35 40 45
Arg Gln Ser Leu Ser Lys Ala Gly Leu Ala Met Asp Ile Val Ala Phe
50 55 60
Asn Gly Glu Cys Ser Gln Gly Glu Val Asp Arg Leu Cys Gln Leu Ala
65 70 75 80
Thr Gln Asn Gly Arg Ser Ala Ile Val Gly Ile Gly Gly Gly Lys Thr
85 90 95
Leu Asp Thr Ala Lys Ala Val Ala Phe Phe Gln Lys Val Pro Val Ala
100 105 110
Val Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser
115 120 125
Val Leu Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Met Leu Pro
130 135 140
Thr Asn Pro Ala Leu Val Val Val Asp Thr Ala Ile Val Ala Arg Ala
145 150 155 160
Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp
165 170 175
Phe Glu Ala Arg Ala Ala Ser Arg Ser Ser Ala Ala Thr Met Ala Gly
180 185 190
Gly Pro Ala Thr Gln Thr Ala Leu Asn Leu Ala Arg Phe Cys Tyr Asp
195 200 205
Thr Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Val Gln Ala Gln
210 215 220
Val Val Thr Pro Ala Leu Glu Arg Ile Val Glu Ala Asn Thr Tyr Leu
225 230 235 240
Ser Gly Val Gly Phe Glu Ser Gly Gly Val Ala Ala Ala His Ala Val
245 250 255
His Asn Gly Leu Thr Ala Val Ala Glu Thr His His Phe Tyr His Gly
260 265 270
Glu Lys Val Ala Phe Gly Val Leu Val Gln Leu Ala Leu Glu Asn Ala
275 280 285
Ser Asn Ala Glu Met Gln Glu Val Met Ser Leu Cys His Ala Val Gly
290 295 300
Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Thr Glu Asp Ile Pro Thr
305 310 315 320
Lys Met Arg Ala Val Ala Glu Leu Ala Cys Ala Pro Gly Glu Thr Ile
325 330 335
His Asn Met Pro Gly Gly Val Thr Val Glu Gln Val Tyr Gly Ala Leu
340 345 350
Leu Val Ala Asp Gln Leu Gly Gln His Phe Leu Glu Phe
355 360 365
<210>80
<211>1104
<212>DNA
<213>凝结芽孢杆菌(Bacillus coagulans)
<220>
<221>CDS
<222>(1)..(1104)
<223>
<400>80
atg acg aaa atc att acc tct cca agc aag ttt ata caa ggc ccc gat 48
Met Thr Lys Ile Ile Thr Ser Pro Ser Lys Phe Ile Gln Gly Pro Asp
1 5 10 15
gaa ttg tcc agg ctt tcg gcg tat acg gaa agg ctt ggc aaa aaa gca 96
Glu Leu Ser Arg Leu Ser Ala Tyr Thr Glu Arg Leu Gly Lys Lys Ala
20 25 30
ttt att att gcg gat gat ttt gtc acc ggc ctt gtc ggc aaa acg gtt 144
Phe Ile Ile Ala Asp Asp Phe Val Thr Gly Leu Val Gly Lys Thr Val
35 40 45
gaa gaa agc tat gcc ggc aaa gaa acg ggg tat caa atg gca tta ttc 192
Glu Glu Ser Tyr Ala Gly Lys Glu Thr Gly Tyr Gln Met Ala Leu Phe
50 55 60
ggt ggt gag tgt tct aaa ccg gaa atc gaa cgg ctt tgt gaa atg agc 240
Gly Gly Glu Cys Ser Lys Pro Glu Ile Glu Arg Leu Cys Glu Met Ser
65 70 75 80
aaa tcc gag gaa gcc gat gtc gtt gtc gga atc ggc ggc gga aaa aca 288
Lys Ser Glu Glu Ala Asp Val Val Val Gly Ile Gly Gly Gly Lys Thr
85 90 95
ttg gat acc gca aaa gca gtc ggg tat tac aat aac att ccg gtg att 336
Leu Asp Thr Ala Lys Ala Val Gly Tyr Tyr Asn Asn Ile Pro Val Ile
100 105 110
gtc gcg ccg acc atc gct tcc acc gat gcc ccg aca agc gcc ctg tct 384
Val Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Thr Ser Ala Leu Ser
115 120 125
gtt att tac aaa gag aac ggc gag ttt gaa gaa tac ttg atg ctg ccg 432
Val Ile Tyr Lys Glu Asn Gly Glu Phe Glu Glu Tyr Leu Met Leu Pro
130 135 140
ctg aac ccg act ttt gtc att atg gat acg aaa gtg att gcc tct gcc 480
Leu Asn Pro Thr Phe Val Ile Met Asp Thr Lys Val Ile Ala Ser Ala
145 150 155 160
cct gcc cgc ctg ctc gtt tcc ggc atg gga gat gcg ctt gca acg tat 528
Pro Ala Arg Leu Leu Val Ser Gly Met Gly Asp Ala Leu Ala Thr Tyr
165 170 175
ttt gaa gcg cgc gcc act aag cgg gca aat aaa acg acg atg gca ggc 576
Phe Glu Ala Arg Ala Thr Lys Arg Ala Asn Lys Thr Thr Met Ala Gly
180 185 190
ggg cgt gtt acg gaa gcg gcg atc gcg ctt gca aaa ctt tgt tat gac 624
Gly Arg Val Thr Glu Ala Ala Ile Ala Leu Ala Lys Leu Cys Tyr Asp
195 200 205
acg caa att tcg gaa ggt tta aaa gca aaa ctg gca gcg gaa aaa cat 672
Thr Gln Ile Ser Glu Gly Leu Lys Ala Lys Leu Ala Ala Glu Lys His
210 215 220
ctt gtt acg gaa gca gtg gaa aaa atc att gaa gcg aat acg tat ctg 720
Leu Val Thr Glu Ala Val Glu Lys Ile Ile Glu Ala Asn Thr Tyr Leu
225 230 235 240
agc gga atc ggt ttt gaa agc ggc ggc ctt gct gcg gca cat gcg atc 768
Ser Gly Ile Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Ile
245 250 255
cat aat ggg ctt acc gtg ctc gaa gaa acc cat cat atg tac cac ggc 816
His Asn Gly Leu Thr Val Leu Glu Glu Thr His His Met Tyr His Gly
260 265 270
gaa aaa gtg gca ttc ggt acc ctc gcc cag ctg att ttg gaa gat gcg 864
Glu Lys Val Ala Phe Gly Thr Leu Ala Gln Leu Ile Leu Glu Asp Ala
275 280 285
ccg aaa gcg gaa att gaa gag gtg gtc tcc ttc tgc ctg agt gtc gga 912
Pro Lys Ala Glu Ile Glu Glu Val Val Ser Phe Cys Leu Ser Val Gly
290 295 300
ctt ccc gtc acg ctc ggg gat ttg ggc gtg aaa gaa ctg aat gag gaa 960
Leu Pro Val Thr Leu Gly Asp Leu Gly Val Lys Glu Leu Asn Glu Glu
305 310 315 320
aag ctc cga aaa gtg gct gaa ctt tcc tgt gcg gaa ggc gaa acg att 1008
Lys Leu Arg Lys Val Ala Glu Leu Ser Cys Ala Glu Gly Glu Thr Ile
325 330 335
tat aac atg ccg ttt gaa gtc acg cct gac ctt gtg tac gca gca atc 1056
Tyr Asn Met Pro Phe Glu Val Thr Pro Asp Leu Val Tyr Ala Ala Ile
340 345 350
gtt acc gct gat tcc gtc ggg cgg tat tat aag gaa aaa tgg gca tga 1104
Val Thr Ala Asp Ser Val Gly Arg Tyr Tyr Lys Glu Lys Trp Ala
355 360 365
<210>81
<211>367
<212>PRT
<213>凝结芽孢杆菌
<400>81
Met Thr Lys Ile Ile Thr Ser Pro Ser Lys Phe Ile Gln Gly Pro Asp
1 5 10 15
Glu Leu Ser Arg Leu Ser Ala Tyr Thr Glu Arg Leu Gly Lys Lys Ala
20 25 30
Phe Ile Ile Ala Asp Asp Phe Val Thr Gly Leu Val Gly Lys Thr Val
35 40 45
Glu Glu Ser Tyr Ala Gly Lys Glu Thr Gly Tyr Gln Met Ala Leu Phe
50 55 60
Gly Gly Glu Cys Ser Lys Pro Glu Ile Glu Arg Leu Cys Glu Met Ser
65 70 75 80
Lys Ser Glu Glu Ala Asp Val Val Val Gly Ile Gly Gly Gly Lys Thr
85 90 95
Leu Asp Thr Ala Lys Ala Val Gly Tyr Tyr Asn AsnIle Pro Val Ile
100 105 110
Val Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Thr Ser Ala Leu Ser
115 120 125
Val Ile Tyr Lys Glu Asn Gly Glu Phe Glu Glu Tyr Leu Met Leu Pro
130 135 140
Leu Asn Pro Thr Phe Val Ile Met Asp Thr Lys Val Ile Ala Ser Ala
145 150 155 160
Pro Ala Arg Leu Leu Val Ser Gly Met Gly Asp Ala Leu Ala Thr Tyr
165 170 175
Phe Glu Ala Arg Ala Thr Lys Arg Ala Asn Lys Thr Thr Met Ala Gly
180 185 190
Gly Arg Val Thr Glu Ala Ala Ile Ala Leu Ala Lys Leu Cys Tyr Asp
195 200 205
Thr Gln Ile Ser Glu Gly Leu Lys Ala Lys Leu Ala Ala Glu Lys His
210 215 220
Leu Val Thr Glu Ala Val Glu Lys Ile Ile Glu Ala Asn Thr Tyr Leu
225 230 235 240
Ser Gly Ile Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Ile
245 250 255
His Asn Gly Leu Thr Val Leu Glu Glu Thr His His Met Tyr His Gly
260 265 270
Glu Lys Val Ala Phe Gly Thr Leu Ala Gln Leu Ile Leu Glu Asp Ala
275 280 285
Pro Lys Ala Glu Ile Glu Glu Val Val Ser Phe Cys Leu Ser Val Gly
290 295 300
Leu Pro Val Thr Leu Gly Asp Leu Gly Val Lys Glu Leu Asn Glu Glu
305 310 315 320
Lys Leu Arg Lys Val Ala Glu Leu Ser Cys Ala Glu Gly Glu Thr Ile
325 330 335
Tyr Asn Met Pro Phe Glu Val Thr Pro Asp Leu Val Tyr Ala Ala Ile
340 345 350
Val Thr Ala Asp Ser Val Gly Arg Tyr Tyr Lys Glu Lys Trp Ala
355 360 365
<210>82
<21l>999
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(999)
<223>
<400>82
atg aaa acg tta ggt gaa ttt att gtc gaa aag cag cac gag ttt tct 48
Met Lys Thr Leu Gly Glu Phe Ile Val Glu Lys Gln His Glu Phe Ser
1 5 10 15
cat gct acc ggt gag ctc act gct ttg ctg tcg gca ata aaa ctg ggc 96
His Ala Thr Gly Glu Leu Thr Ala Leu Leu Ser Ala Ile Lys Leu Gly
20 25 30
gcc aag att atc cat cgc gat atc aac aaa gca gga ctg gtt gat atc 144
Ala Lys Ile Ile His Arg Asp Ile Asn Lys Ala Gly Leu Val Asp Ile
35 40 45
ctg ggt gcc agc ggt gct gag aac gtg cag ggc gag gtt cag cag aaa 192
Leu Gly Ala Ser Gly Ala Glu Asn Val Gln Gly Glu Val Gln Gln Lys
50 55 60
ctc gac ttg ttc gct aat gaa aaa ctg aaa gcc gca ctg aaa gca cgc 240
Leu Asp Leu Phe Ala Asn Glu Lys Leu Lys Ala Ala Leu Lys Ala Arg
65 70 75 80
gat atc gtt gcg ggc att gcc tct gaa gaa gaa gat gag att gtc gtc 288
Asp Ile Val Ala Gly Ile Ala Ser Glu Glu Glu Asp Glu Ile Val Val
85 90 95
ttt gaa ggc tgt gaa cac gca aaa tac gtg gtg ctg atg gac ccc ctg 336
Phe Glu Gly Cys Glu His Ala Lys Tyr Val Val Leu Met Asp Pro Leu
100 105 110
gat ggc tcg tcc aac atc gat gtt aac gtc tct gtc ggt acc att ttc 384
Asp Gly Ser Ser Asn Ile Asp Val Asn Val Ser Val Gly Thr Ile Phe
115 120 125
tcc atc tac cgc cgc gtt acg cct gtt ggc acg ccg gta acg gaa gaa 432
Ser Ile Tyr Arg Arg Val Thr Pro Val Gly Thr Pro Val Thr Glu Glu
130 135 140
gat ttc ctc cag cct ggt aac aaa cag gtt gcg gca ggt tac gtg gta 480
Asp Phe Leu Gln Pro Gly Asn Lys Gln Val Ala Ala Gly Tyr Val Val
145 150 155 160
tac ggc tcc tct acc atg ctg gtt tac acc acc gga tgc ggt gtt cac 528
Tyr Gly Ser Ser Thr Met Leu Val Tyr Thr Thr Gly Cys Gly Val His
165 170 175
gcc ttt act tac gat cct tcg ctc ggc gtt ttc tgc ctg tgc cag gaa 576
Ala Phe Thr Tyr Asp Pro Ser Leu GIy Val Phe Cys Leu Cys Gln Glu
180 185 190
cgg atg cgc ttc ccg gag aaa ggc aaa acc tac tcc atc aac gaa gga 624
Arg Met Arg Phe Pro Glu Lys Gly Lys Thr Tyr Ser Ile Asn Glu Gly
195 200 205
aac tac att aag ttt ccg aac ggg gtg aag aag tac att aaa ttc tgc 672
Asn Tyr Ile Lys Phe Pro Asn Gly Val Lys Lys Tyr Ile Lys Phe Cys
210 215 220
cag gaa gaa gat aaa tcc acc aac cgc cct tat acc tca cgt tat atc 720
Gln Glu Glu Asp Lys Ser Thr Asn Arg Pro Tyr Thr Ser Arg Tyr Ile
225 230 235 240
ggt tca ctg gtc gcg gat ttc cac cgt aac ctg ctg aaa ggc ggt att 768
Gly Ser Leu Val Ala Asp Phe His Arg Asn Leu Leu Lys Gly Gly Ile
245 250 255
tat ctc tac cca agc acc gcc agc cac ccg gac ggc aaa ctg cgt ttg 816
Tyr Leu Tyr Pro Ser Thr Ala Ser His Pro Asp Gly Lys Leu Arg Leu
260 265 270
ctg tat gag tgc aac ccg atg gca ttc ctg gcg gaa caa gcg ggc ggt 864
Leu Tyr Glu Cys Asn Pro Met Ala Phe Leu Ala Glu Gln Ala Gly Gly
275 280 285
aaa gcg agc gat ggc aaa gag cgt att ctg gat atc atc ccg gaa acc 912
Lys Ala Ser Asp Gly Lys Glu Arg Ile Leu Asp Ile Ile Pro Glu Thr
290 295 300
ctg cac cag cgc cgt tca ttc ttt gtc ggc aac gac cat atg gtt gaa 960
Leu His Gln Arg Arg Ser Phe Phe Val Gly Asn Asp His Met Val Glu
305 310 315 320
gat gtc gaa cgc ttt atc cgt gag ttc ccg gac gcg taa 999
Asp Val Glu Arg Phe Ile Arg Glu Phe Pro Asp Ala
325 330
<210>83
<211>332
<212>PRT
<213>大肠杆菌
<400>83
Met Lys Thr Leu Gly Glu Phe Ile Val Glu Lys Gln His Glu Phe Ser
1 5 10 15
His Ala Thr Gly Glu Leu Thr Ala Leu Leu Ser Ala Ile Lys Leu Gly
20 25 30
Ala Lys Ile Ile His Arg Asp Ile Asn Lys Ala Gly Leu Val Asp Ile
35 40 45
Leu Gly Ala Ser Gly Ala Glu Asn Val Gln Gly Glu Val Gln Gln Lys
50 55 60
Leu Asp Leu Phe Ala Asn Glu Lys Leu Lys Ala Ala Leu Lys Ala Arg
65 70 75 80
Asp Ile Val Ala Gly Ile Ala Ser Glu Glu Glu Asp Glu Ile Val Val
85 90 95
Phe Glu Gly Cys Glu His Ala Lys Tyr Val Val Leu Met Asp Pro Leu
100 105 110
Asp Gly Ser Ser Asn Ile Asp Val Asn Val Ser Val Gly Thr Ile Phe
115 120 125
Ser Ile Tyr Arg Arg Val Thr Pro Val Gly Thr Pro Val Thr Glu Glu
130 135 140
Asp Phe Leu Gln Pro Gly Asn Lys Gln Val Ala Ala Gly Tyr Val Val
145 150 155 160
Tyr Gly Ser Ser Thr Met Leu Val Tyr Thr Thr Gly Cys Gly Val His
165 170 175
Ala Phe Thr Tyr Asp Pro Ser Leu Gly Val Phe Cys Leu Cys Gln Glu
180 185 190
Arg Met Arg Phe Pro Glu Lys Gly Lys Thr Tyr Ser Ile Asn Glu Gly
195 200 205
Asn Tyr Ile Lys Phe Pro Asn Gly Val Lys Lys Tyr Ile Lys Phe Cys
210 215 220
Gln Glu Glu Asp Lys Ser Thr Asn Arg Pro Tyr Thr Ser Arg Tyr Ile
225 230 235 240
Gly Ser Leu Val Ala Asp Phe His Arg Asn Leu Leu Lys Gly Gly Ile
245 250 255
Tyr Leu Tyr Pro Ser Thr Ala Ser His Pro Asp Gly Lys Leu Arg Leu
260 265 270
Leu Tyr Glu Cys Asn Pro Met Ala Phe Leu Ala Glu Gln Ala Gly Gly
275 280 285
Lys Ala Ser Asp Gly Lys Glu Arg Ile Leu Asp Ile Ile Pro Glu Thr
290 295 300
Leu His Gln Arg Arg Ser Phe Phe Val Gly Asn Asp His Met Val Glu
305 310 315 320
Asp Val Glu Arg Phe Ile Arg Glu Phe Pro Asp Ala
325 330
<210>84
<211>819
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(819)
<223>
<400>84
atg acc aca cgc gtg att gct ctc gac tta gac ggc acc tta ttg acc 48
Met Thr Thr Arg Val Ile Ala Leu Asp Leu Asp Gly Thr Leu Leu Thr
1 5 10 15
ccg aaa aag acc ctg ctt cct tca tcg ata gaa gcc ctg gcc cgc gct 96
Pro Lys Lys Thr Leu Leu Pro Ser Ser Ile Glu Ala Leu Ala Arg Ala
20 25 30
cgc gaa gca ggc tat caa tta atc atc gtc aca ggt cgc cat cac gtc 144
Arg G1u Ala Gly Tyr Gln Leu Ile Ile Val Thr Gly Arg His His Val
35 40 45
gct att cat cct ttt tat cag gcg ctg gcg ctg gat aca cct gct att 192
Ala Ile His Pro Phe Tyr Gln Ala Leu Ala Leu Asp Thr Pro Ala Ile
50 55 60
tgc tgt aat ggc acc tat ttg tat gat tat cat gca aaa acc gtg ctg 240
Cys Cys Asn Gly Thr Tyr Leu Tyr Asp Tyr His Ala Lys Thr Val Leu
65 70 75 80
gaa gcg gac cca atg ccc gtt att aaa gcc ctg caa ctc att gag atg 288
Glu Ala Asp Pro Met Pro Val Ile Lys Ala Leu Gln Leu Ile Glu Met
85 90 95
ctg aat gaa cac cac att cac ggt ctg atg tat gtc gat gat gca atg 336
Leu Asn Glu His His Ile His Gly Leu Met Tyr Val Asp Asp Ala Met
100 105 110
gtc tat gag cac ccg acc ggg cat gtc att cgc aca tct aac tgg gcg 384
Val Tyr Glu His Pro Thr Gly His Val Ile Arg Thr Ser Asn Trp Ala
115 120 125
caa acc ctg ccg ccg gaa cag cgt ccg act ttc aca caa gtc gct tct 432
Gln Thr Leu Pro Pro Glu Gln Arg Pro Thr Phe Thr Gln Val Ala Ser
130 135 140
ctg gct gaa acg gcg caa caa gtt aac gcc gta tgg aag ttc gcc ctg 480
Leu Ala Glu Thr Ala Gln Gln Val Asn Ala Val Trp Lys Phe Ala Leu
145 150 155 160
acg cac gat gac ctg ccg caa ttg cag cat ttt ggt aag cat gtc gaa 528
Thr His Asp Asp Leu Pro Gln Leu Gln His Phe Gly Lys His Val Glu
165 170 175
cat gaa ctg gga ctg gag tgt gaa tgg tcc tgg cac gat cag gtt gat 576
His Glu Leu Gly Leu Glu Cys Glu Trp Ser Trp His Asp Gln Val Asp
180 185 190
att gca cgc ggc ggc aac agc aaa ggt aaa cgt ttg acg aaa tgg gtt 624
Ile Ala Arg Gly Gly Asn Ser Lys Gly Lys Arg Leu Thr Lys Trp Val
195 200 205
gag gcg caa ggt tgg tcg atg gaa aac gtc gtg gca ttc ggc gat aac 672
Glu Ala Gln Gly Trp Ser Met Glu Asn Val Val Ala Phe Gly Asp Asn
210 215 220
ttt aat gat atc agt atg ctg gaa gcc gct ggt aca ggc gtg gcg atg 720
Phe Asn Asp Ile Ser Met Leu Glu Ala Ala Gly Thr Gly Val Ala Met
225 230 235 240
ggc aac gcc gat gac gcg gta aaa gcg cgc gcc aac att gtg att ggt 768
Gly Asn Ala Asp Asp Ala Val Lys Ala Arg Ala Asn Ile Val Ile Gly
245 250 255
gat aac acc acc gac agc att gcc cag ttc att tat agc cac ctg att 816
Asp Asn Thr Thr Asp Ser Ile Ala Gln Phe Ile Tyr Ser His Leu Ile
260 265 270
taa 819
<210>85
<211>272
<212>PRT
<213>大肠杆菌
<400>85
Met Thr Thr Arg Val Ile Ala Leu Asp Leu Asp Gly Thr Leu Leu Thr
1 5 10 15
Pro Lys Lys Thr Leu Leu Pro Ser Ser Ile Glu Ala Leu Ala Arg Ala
20 25 30
Arg Glu Ala Gly Tyr Gln Leu Ile Ile Val Thr Gly Arg His His Val
35 40 45
Ala Ile His Pro Phe Tyr Gln Ala Leu Ala Leu Asp Thr Pro Ala Ile
50 55 60
Cys Cys Asn Gly Thr Tyr Leu Tyr Asp Tyr His Ala Lys Thr Val Leu
65 70 75 80
Glu Ala Asp Pro Met Pro Val Ile Lys Ala Leu Gln Leu Ile Glu Met
85 90 95
Leu Asn Glu His His Ile His Gly Leu Met Tyr Val Asp Asp Ala Met
100 105 110
Val Tyr Glu His Pro Thr Gly His Val Ile Arg Thr Ser Asn Trp Ala
115 120 125
Gln Thr Leu Pro Pro Glu Gln Arg Pro Thr Phe Thr Gln Val Ala Ser
130 135 140
Leu Ala Glu Thr Ala Gln Gln Val Asn Ala Val Trp Lys Phe Ala Leu
145 150 155 160
Thr His Asp Asp Leu Pro Gln Leu Gln His Phe Gly Lys His Val Glu
165 170 175
His Glu Leu Gly Leu Glu Cys Glu Trp Ser Trp His Asp Gln Val Asp
180 185 190
Ile Ala Arg Gly Gly Asn Ser Lys Gly Lys Arg Leu Thr Lys Trp Val
195 200 205
Glu Ala Gln Gly Trp Ser Met Glu Asn Val Val Ala Phe Gly Asp Asn
210 215 220
Phe Asn Asp Ile Ser Met Leu Glu Ala Ala Gly Thr Gly Val Ala Met
225 230 235 240
Gly Asn Ala Asp Asp Ala Val Lys Ala Arg Ala Asn Ile Val Ile Gly
245 250 255
Asp Asn Thr Thr Asp Ser Ile Ala Gln Phe Ile Tyr Ser His Leu Ile
260 265 270
<210>86
<211>1728
<212>DNA
<213>大肠杆菌
<220>
<221>CDS
<222>(1)..(1728)
<223>
<400>86
atg att tca ggc att tta gca tcc ccg ggt atc gct ttc ggt aaa gct 48
Met Ile Ser Gly Ile Leu Ala Ser Pro Gly Ile Ala Phe Gly Lys Ala
1 5 10 15
ctg ctt ctg aaa gaa gac gaa att gtc att gac cgg aaa aaa att tct 96
Leu Leu Leu Lys Glu Asp Glu Ile Val Ile Asp Arg Lys Lys Ile Ser
20 25 30
gcc gac cag gtt gat cag gaa gtt gaa cgt ttt ctg agc ggt cgt gcc 144
Ala Asp Gln Val Asp Gln Glu Val Glu Arg Phe Leu Ser Gly Arg Ala
35 40 45
aag gca tca gcc cag ctg gaa acg atc aaa acg aaa gct ggt gaa acg 192
Lys Ala Ser Ala Gln Leu Glu Thr Ile Lys Thr Lys Ala Gly Glu Thr
50 55 60
ttc ggt gaa gaa aaa gaa gcc atc ttt gaa ggg cat att atg ctg ctc 240
Phe Gly Glu Glu Lys Glu Ala Ile Phe Glu Gly His Ile Met Leu Leu
65 70 75 80
gaa gat gag gag ctg gag cag gaa atc ata gcc ctg att aaa gat aag 288
Glu Asp Glu Glu Leu Glu Gln Glu Ile Ile Ala Leu Ile Lys Asp Lys
85 90 95
cac atg aca gct gac gca gct gct cat gaa gtt atc gaa ggt cag gct 336
His Met Thr Ala Asp Ala Ala Ala His Glu Val Ile Glu Gly Gln Ala
100 105 110
tct gcc ctg gaa gag ctg gat gat gaa tac ctg aaa gaa cgt gcg gct 384
Ser Ala Leu Glu Glu Leu Asp Asp Glu Tyr Leu Lys Glu Arg Ala Ala
115 120 125
gac gta cgt gat atc ggt aag cgc ctg ctg cgc aac atc ctg ggc ctg 432
Asp Val Arg Asp Ile Gly Lys Arg Leu Leu Arg Asn Ile Leu Gly Leu
130 135 140
aag att atc gac ctg agc gcc att cag gat gaa gtc att ctg gtt gcc 480
Lys Ile Ile Asp Leu Ser Ala Ile Gln Asp Glu Val Ile Leu Val Ala
145 150 155 160
gct gac ctg acg ccg tcc gaa acc gca cag ctg aac ctg aag aag gtg 528
Ala Asp Leu Thr Pro Ser Glu Thr Ala Gln Leu Asn Leu Lys Lys Val
165 170 175
ctg ggt ttc atc acc gac gcg ggt ggc cgt act tcc cac acc tct atc 576
Leu Gly Phe Ile Thr Asp Ala Gly Gly Arg Thr Ser His Thr Ser Ile
180 185 190
atg gcg cgt tct ctg gaa cta cct gct atc gtg ggt acc ggt agc gtc 624
Met Ala Arg Ser Leu Glu Leu Pro Ala Ile Val Gly Thr Gly Ser Val
195 200 205
acc tct cag gtg aaa aat gac gac tat ctg att ctg gat gcc gta aat 672
Thr Ser Gln Val Lys Asn Asp Asp Tyr Leu Ile Leu Asp Ala Val Asn
210 215 220
aat cag gtt tac gtc aat cca acc aac gaa gtt att gat aaa atg cgc 720
Asn Gln Val Tyr Val Asn Pro Thr Asn Glu Val Ile Asp Lys Met Arg
225 230 235 240
gct gtt cag gag caa gtg gct tct gaa aaa gca gag ctt gct aaa ctg 768
Ala Val Gln Glu Gln Val Ala Ser Glu Lys Ala Glu Leu Ala Lys Leu
245 250 255
aaa gat ctg cca gct att acg ctg gac ggt cac cag gta gaa gta tgc 816
Lys Asp Leu Pro Ala Ile Thr Leu Asp Gly His Gln Val Glu Val Cys
260 265 270
gct aac att ggt acg gtt cgt gac gtt gaa ggt gca gag cgt aac ggc 864
Ala Asn Ile Gly Thr Val Arg Asp Val Glu Gly Ala Glu Arg Asn Gly
275 280 285
gct gaa ggc gtt ggt ctg tat cgt act gag ttc ctg ttc atg gac cgc 912
Ala Glu Gly Val Gly Leu Tyr Arg Thr Glu Phe Leu Phe Met Asp Arg
290 295 300
gac gca ctg ccc act gaa gaa gaa cag ttt gct gct tac aaa gca gtg 960
Asp Ala Leu Pro Thr Glu Glu Glu Gln Phe Ala Ala Tyr Lys Ala Val
305 310 315 320
gct gaa gcg tgt ggc tcg caa gcg gtt atc gtt cgt acc atg gac atc 1008
Ala Glu Ala Cys Gly Ser Gln Ala Val Ile Val Arg Thr Met Asp Ile
325 330 335
ggc ggc gac aaa gag ctg cca tac atg aac ttc ccg aaa gaa gag aac 1056
Gly Gly Asp Lys Glu Leu Pro Tyr Met Asn Phe Pro Lys Glu Glu Asn
340 345 350
ccg ttc ctc ggc tgg cgc gct atc cgt atc gcg atg gat cgt aga gag 1104
Pro Phe Leu Gly Trp Arg Ala Ile Arg Ile Ala Met Asp Arg Arg Glu
355 360 365
atc ctg cgc gat cag ctc cgc gct atc ctg cgt gcc tcg gct ttc ggt 1152
Ile Leu Arg Asp Gln Leu Arg Ala Ile Leu Arg Ala Ser Ala Phe Gly
370 375 380
aaa ttg cgc att atg ttc ccg atg atc atc tct gtt gaa gaa gtg cgt 1200
Lys Leu Arg Ile Met Phe Pro Met Ile Ile Ser Val Glu Glu Val Arg
385 390 395 400
gca ctg cgc aaa gag atc gaa atc tac aaa cag gaa ctg cgc gac gaa 1248
Ala Leu Arg Lys Glu Ile Glu Ile Tyr Lys Gln Glu Leu Arg Asp Glu
405 410 415
ggt aaa gcg ttt gac gag tca att gaa atc ggc gta atg gtg gaa aca 1296
Gly Lys Ala Phe Asp Glu Ser Ile Glu Ile Gly Val Met Val Glu Thr
420 425 430
ccg gct gcc gca aca att gca cgt cat tta gcc aaa gaa gtt gat ttc 1344
Pro Ala Ala Ala Thr Ile Ala Arg His Leu Ala Lys Glu Val Asp Phe
435 440 445
ttt agt atc ggc acc aat gat tta acg cag tac act ctg gca gtt gac 1392
Phe Ser Ile Gly Thr Asn Asp Leu Thr Gln Tyr Thr Leu Ala Val Asp
450 455 460
cgt ggt aat gat atg att tca cac ctt tac cag cca atg tca ccg tcc 1440
Arg Gly Asn Asp Met Ile Ser His Leu Tyr Gln Pro Met Ser Pro Ser
465 470 475 480
gtg ctg aac ttg atc aag caa gtt att gat gct tct cat gct gaa ggc 1488
Val Leu Asn Leu Ile Lys Gln Val Ile Asp Ala Ser His Ala Glu Gly
485 490 495
aaa tgg act ggc atg tgt ggt gag ctt gct ggc gat gaa cgt gct aca 1536
Lys Trp Thr Gly Met Cys Gly Glu Leu Ala Gly Asp Glu Arg Ala Thr
500 505 510
ctt ctg ttg ctg ggg atg ggt ctg gac gaa ttc tct atg agc gcc att 1584
Leu Leu Leu Leu Gly Met Gly Leu Asp Glu Phe Ser Met Ser Ala Ile
515 520 525
tct atc ccg cgc att aag aag att atc cgt aac acg aac ttc gaa gat 1632
Ser Ile Pro Arg Ile Lys Lys Ile Ile Arg Asn Thr Asn Phe Glu Asp
530 535 540
gcg aag gtg tta gca gag cag gct ctt gct caa ccg aca acg gac gag 1680
Ala Lys Val Leu Ala Glu Gln Ala Leu Ala Gln Pro Thr Thr Asp Glu
545 550 555 560
tta atg acg ctg gtt aac aag ttc att gaa gaa aaa aca atc tgc taa 1728
Leu Met Thr Leu Val Asn Lys Phe Ile Glu Glu Lys Thr Ile Cys
565 570 575
<210>87
<211>575
<212>PRT
<213>大肠杆菌
<400>87
Met Ile Ser Gly Ile Leu Ala Ser Pro Gly Ile Ala Phe Gly Lys Ala
1 5 10 15
Leu Leu Leu Lys Glu Asp Glu Ile Val Ile Asp Arg Lys Lys Ile Ser
20 25 30
Ala Asp Gln Val Asp Gln Glu Val Glu Arg Phe Leu Ser Gly Arg Ala
35 40 45
Lys Ala Ser Ala Gln Leu Glu Thr Ile Lys Thr Lys Ala Gly Glu Thr
50 55 60
Phe Gly Glu Glu Lys Glu Ala Ile Phe Glu Gly His Ile Met Leu Leu
65 70 75 80
Glu Asp Glu Glu Leu Glu Gln Glu Ile Ile Ala Leu Ile Lys Asp Lys
85 90 95
His Met Thr Ala Asp Ala Ala Ala His Glu Val Ile Glu Gly Gln Ala
100 105 110
Ser Ala Leu Glu Glu Leu Asp Asp Glu Tyr Leu Lys Glu Arg Ala Ala
115 120 125
Asp Val Arg Asp Ile Gly Lys Arg Leu Leu Arg Asn Ile Leu Gly Leu
130 135 140
Lys Ile Ile Asp Leu Ser Ala Ile Gln Asp Glu Val Ile Leu Val Ala
145 150 155 160
Ala Asp Leu Thr Pro Ser Glu Thr Ala Gln Leu Asn Leu Lys Lys Val
165 170 175
Leu Gly Phe Ile Thr Asp Ala Gly Gly Arg Thr Ser His Thr Ser Ile
180 185 190
Met Ala Arg Ser Leu Glu Leu Pro Ala Ile Val Gly Thr Gly Ser Val
195 200 205
Thr Ser Gln Val Lys Asn Asp Asp Tyr Leu Ile Leu Asp Ala Val Asn
210 215 220
Asn Gln Val Tyr Val Asn Pro Thr Asn Glu Val Ile Asp Lys Met Arg
225 230 235 240
Ala Val Gln Glu Gln Val Ala Ser Glu Lys Ala Glu Leu Ala Lys Leu
245 250 255
Lys Asp Leu Pro Ala Ile Thr Leu Asp Gly His Gln Val Glu Val Cys
260 265 270
Ala Asn Ile Gly Thr Val Arg Asp Val Glu Gly Ala Glu Arg Asn Gly
275 280 285
Ala Glu Gly Val Gly Leu Tyr Arg Thr Glu Phe Leu Phe Met Asp Arg
290 295 300
Asp Ala Leu Pro Thr Glu Glu Glu Gln Phe Ala Ala Tyr Lys Ala Val
305 310 315 320
Ala Glu Ala Cys Gly Ser Gln Ala Val Ile Val Arg Thr Met Asp Ile
325 330 335
Gly Gly Asp Lys Glu Leu Pro Tyr Met Asn Phe Pro Lys Glu Glu Asn
340 345 350
Pro Phe Leu Gly Trp Arg Ala Ile Arg Ile Ala Met Asp Arg Arg Glu
355 360 365
Ile Leu Arg Asp Gln Leu Arg Ala Ile Leu Arg Ala Ser Ala Phe Gly
370 375 380
Lys Leu Arg Ile Met Phe Pro Met Ile Ile Ser Val Glu Glu Val Arg
385 390 395 400
Ala Leu Arg Lys Glu Ile Glu Ile Tyr Lys Gln Glu Leu Arg Asp Glu
405 410 415
Gly Lys Ala Phe Asp Glu Ser Ile Glu Ile Gly Val Met Val Glu Thr
420 425 430
Pro Ala Ala Ala Thr Ile Ala Arg His Leu Ala Lys Glu Val Asp Phe
435 440 445
Phe Ser Ile Gly Thr Asn Asp Leu Thr Gln Tyr Thr Leu Ala Val Asp
450 455 460
Arg Gly Asn Asp Met Ile Ser His Leu Tyr Gln Pro Met Ser Pro Ser
465 470 475 480
Val Leu Asn Leu Ile Lys Gln Val Ile Asp Ala Ser His Ala Glu Gly
485 490 495
Lys Trp Thr Gly Met Cys Gly Glu Leu Ala Gly Asp Glu Arg Ala Thr
500 505 510
Leu Leu Leu Leu Gly Met Gly Leu Asp Glu Phe Ser Met Ser Ala Ile
515 520 525
Ser Ile Pro Arg Ile Lys Lys Ile Ile Arg Asn Thr Asn Phe Glu Asp
530 535 540
Ala Lys Val Leu Ala Glu Gln Ala Leu Ala Gln Pro Thr Thr Asp Glu
545 550 555 560
Leu Met Thr Leu Val Asn Lys Phe Ile Glu Glu Lys Thr Ile Cys
565 570 575
<210>88
<211>1740
<212>DNA
<213>人工序列
<220>
<223>attR-cat-attL-PtacM2-SD-spacer
<400>88
tctagacgct caagttagta taaaaaagct gaacgagaaa cgtaaaatga tataaatatc 60
aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat 120
atgcagtcac tatgaatcaa ctacttagat ggtattagtg acctgtaaca gactgcagtg 180
gtcgaaaaaa aaagcccgca ctgtcaggtg cgggcttttt tctgtgttaa gcttcgacga 240
atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg 300
caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt 360
aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc 420
gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg 480
gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga 540
ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca 600
ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat 660
tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga 720
acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca 780
ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt 840
acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca 900
actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta 960
tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cggatccggc 1020
caagctagct tggctctagc tagagcgccc ggttgacgct gctagtgtta cctagcgatt 1080
tgtatcttac tgcatgttac ttcatgttgt caatacctgt ttttcgtgcg acttatcagg 1140
ctgtctactt atccggagat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata 1200
gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct 1260
ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag 1320
tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata 1380
accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg 1440
ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat 1500
taaagcttat cgatgataag ctgtcaaaca tgagaattcg aaatcaaata atgattttat 1560
tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt tttataatgc 1620
caacttagta taaaaaagca ggcttcaaga tctctcccca tccccctgtg tacaattaat 1680
catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca ggagactgcc 1740
<210>89
<211>1740
<212>DNA
<213>人工序列
<220>
<223>attR-cat-attL-PtacM3-SD-spacer
<400>89
tctagacgct caagttagta taaaaaagct gaacgagaaa cgtaaaatga tataaatatc 60
aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat 120
atgcagtcac tatgaatcaa ctacttagat ggtattagtg acctgtaaca gactgcagtg 180
gtcgaaaaaa aaagcccgca ctgtcaggtg cgggcttttt tctgtgttaa gcttcgacga 240
atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg 300
caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt 360
aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc 420
gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg 480
gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga 540
ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca 600
ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat 660
tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga 720
acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca 780
ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt 840
acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca 900
actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta 960
tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cggatccggc 1020
caagctagct tggctctagc tagagcgccc ggttgacgct gctagtgtta cctagcgatt 1080
tgtatcttac tgcatgttac ttcatgttgt caatacctgt ttttcgtgcg acttatcagg 1140
ctgtctactt atccggagat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata 1200
gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct 1260
ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag 1320
tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata 1380
accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg 1440
ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat 1500
taaagcttat cgatgataag ctgtcaaaca tgagaattcg aaatcaaata atgattttat 1560
tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt tttataatgc 1620
caacttagta taaaaaagca ggcttcaaga tctctcccca tccccctgtt ggcaattaat 1680
catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca ggagactgcc 1740
<210>90
<211>2847
<212>DNA
<213>人工序列
<220>
<223>PtacM2gldA::Cm
<400>90
tctagacgct caagttagta taaaaaagct gaacgagaaa cgtaaaatga tataaatatc 60
aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat 120
atgcagtcac tatgaatcaa ctacttagat ggtattagtg acctgtaaca gactgcagtg 180
gtcgaaaaaa aaagcccgca ctgtcaggtg cgggcttttt tctgtgttaa gcttcgacga 240
atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg 300
caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt 360
aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc 420
gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg 480
gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga 540
ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca 600
ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat 660
tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga 720
acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca 780
ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt 840
acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca 900
actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta 960
tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cggatccggc 1020
caagctagct tggctctagc tagagcgccc ggttgacgct gctagtgtta cctagcgatt 1080
tgtatcttac tgcatgttac ttcatgttgt caatacctgt ttttcgtgcg acttatcagg 1140
ctgtctactt atccggagat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata 1200
gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct 1260
ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag 1320
tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata 1380
accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg 1440
ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat 1500
taaagcttat cgatgataag ctgtcaaaca tgagaattcg aaatcaaata atgattttat 1560
tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt tttataatgc 1620
caacttagta taaaaaagca ggcttcaaga tcttctctcc ccatccccct gtgtacaatt 1680
aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggagact 1740
gccatggacc gcattattca atcaccgggt aaatacatcc agggcgctga tgtgattaat 1800
cgtctgggcg aatacctgaa gccgctggca gaacgctggt tagtggtggg tgacaaattt 1860
gttttaggtt ttgctcaatc cactgtcgag aaaagcttta aagatgctgg actggtagta 1920
gaaattgcgc cgtttggcgg tgaatgttcg caaaatgaga tcgaccgtct gcgtggcatc 1980
gcggagactg cgcagtgtgg cgcaattctc ggtatcggtg gcggaaaaac cctcgatact 2040
gccaaagcac tggcacattt catgggtgtt ccggtagcga tcgcaccgac tatcgcctct 2100
accgatgcac cgtgcagcgc attgtctgtt atctacaccg atgagggtga gtttgaccgc 2160
tatctgctgt tgccaaataa cccgaatatg gtcattgtcg acaccaaaat cgtcgctggc 2220
gcacctgcac gtctgttagc ggcgggtatc ggcgatgcgc tggcaacctg gtttgaagcg 2280
cgtgcctgct ctcgtagcgg cgcgaccacc atggcgggcg gcaagtgcac ccaggctgcg 2340
ctggcactgg ctgaactgtg ctacaacacc ctgctggaag aaggcgaaaa agcgatgctt 2400
gctgccgaac agcatgtagt gactccggcg ctggagcgcg tgattgaagc gaacacctat 2460
ttgagcggtg ttggttttga aagtggtggt ctggctgcgg cgcacgcagt gcataacggc 2520
ctgaccgcta tcccggacgc gcatcactat tatcacggtg aaaaagtggc attcggtacg 2580
ctgacgcagc tggttctgga aaatgcgccg gtggaggaaa tcgaaaccgt agctgccctt 2640
agccatgcgg taggtttgcc aataactctc gctcaactgg atattaaaga agatgtcccg 2700
gcgaaaatgc gaattgtggc agaagcggca tgtgcagaag gtgaaaccat tcacaacatg 2760
cctggcggcg cgacgccaga tcaggtttac gccgctctgc tggtagccga ccagtacggt 2820
cagcgtttcc tgcaagagtg ggaataa 2847
<210>91
<211>2847
<212>DNA
<213>人工序列
<220>
<223>PtacM3gldA::Cm
<400>91
tctagacgct caagttagta taaaaaagct gaacgagaaa cgtaaaatga tataaatatc 60
aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat 120
atgcagtcac tatgaatcaa ctacttagat ggtattagtg acctgtaaca gactgcagtg 180
gtcgaaaaaa aaagcccgca ctgtcaggtg cgggcttttt tctgtgttaa gcttcgacga 240
atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg 300
caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt 360
aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc 420
gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg 480
gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga 540
ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca 600
ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat 660
tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga 720
acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca 780
ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt 840
acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca 900
actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta 960
tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cggatccggc 1020
caagctagct tggctctagc tagagcgccc ggttgacgct gctagtgtta cctagcgatt 1080
tgtatcttac tgcatgttac ttcatgttgt caatacctgt ttttcgtgcg acttatcagg 1140
ctgtctactt atccggagat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata 1200
gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct 1260
ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag 1320
tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata 1380
accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg 1440
ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat 1500
taaagcttat cgatgataag ctgtcaaaca tgagaattcg aaatcaaata atgattttat 1560
tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt tttataatgc 1620
caacttagta taaaaaagca ggcttcaaga tcttctctcc ccatccccct gttggcaatt 1680
aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggagact 1740
gccatggacc gcattattca atcaccgggt aaatacatcc agggcgctga tgtgattaat 1800
cgtctgggcg aatacctgaa gccgctggca gaacgctggt tagtggtggg tgacaaattt 1860
gttttaggtt ttgctcaatc cactgtcgag aaaagcttta aagatgctgg actggtagta 1920
gaaattgcgc cgtttggcgg tgaatgttcg caaaatgaga tcgaccgtct gcgtggcatc 1980
gcggagactg cgcagtgtgg cgcaattctc ggtatcggtg gcggaaaaac cctcgatact 2040
gccaaagcac tggcacattt catgggtgtt ccggtagcga tcgcaccgac tatcgcctct 2100
accgatgcac cgtgcagcgc attgtctgtt atctacaccg atgagggtga gtttgaccgc 2160
tatctgctgt tgccaaataa cccgaatatg gtcattgtcg acaccaaaat cgtcgctggc 2220
gcacctgcac gtctgttagc ggcgggtatc ggcgatgcgc tggcaacctg gtttgaagcg 2280
cgtgcctgct ctcgtagcgg cgcgaccacc atggcgggcg gcaagtgcac ccaggctgcg 2340
ctggcactgg ctgaactgtg ctacaacacc ctgctggaag aaggcgaaaa agcgatgctt 2400
gctgccgaac agcatgtagt gactccggcg ctggagcgcg tgattgaagc gaacacctat 2460
ttgagcggtg ttggttttga aagtggtggt ctggctgcgg cgcacgcagt gcataacggc 2520
ctgaccgcta tcccggacgc gcatcactat tatcacggtg aaaaagtggc attcggtacg 2580
ctgacgcagc tggttctgga aaatgcgccg gtggaggaaa tcgaaaccgt agctgccctt 2640
agccatgcgg taggtttgcc aataactctc gctcaactgg atattaaaga agatgtcccg 2700
gcgaaaatgc gaattgtggc agaagcggca tgtgcagaag gtgaaaccat tcacaacatg 2760
cctggcggcg cgacgccaga tcaggtttac gccgctctgc tggtagccga ccagtacggt 2820
cagcgtttcc tgcaagagtg ggaataa 2847
<210>92
<211>2903
<212>DNA
<213>人工序列
<220>
<223>PtacM fsaB-gldA::Cm
<400>92
tctagacgct caagttagta taaaaaagct gaacgagaaa cgtaaaatga tataaatatc 60
aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat 120
atgcagtcac tatgaatcaa ctacttagat ggtattagtg acctgtaaca gactgcagtg 180
gtcgaaaaaa aaagcccgca ctgtcaggtg cgggcttttt tctgtgttaa gcttcgacga 240
atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg cgtttaaggg 300
caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt 360
aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc 420
gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg 480
gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga 540
ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca 600
ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat 660
tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga 720
acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca 780
ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt 840
acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca 900
actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta 960
tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cggatccggc 1020
caagctagct tggctctagc tagagcgccc ggttgacgct gctagtgtta cctagcgatt 1080
tgtatcttac tgcatgttac ttcatgttgt caatacctgt ttttcgtgcg acttatcagg 1140
ctgtctactt atccggagat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata 1200
gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct 1260
ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag 1320
tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata 1380
accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg 1440
ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat 1500
taaagcttat cgatgataag ctgtcaaaca tgagaattcg aaatcaaata atgattttat 1560
tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt tttataatgc 1620
caacttagta taaaaaagca ggcttcaaga tcttctctcc ccatccccct gttcacaatt 1680
aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggacatc 1740
atggaactgt atctggacac cgctaacgtc gcagaagtcg aacgtctggc acgcatattc 1800
cccattgccg gggtgacaac taacccgagc attatcgctg ccagcaagga gtccatatgg 1860
gaagtgctgc cgcgtctgca aaaagcgatt ggtgatgagg gcattctgtt tgctcagacc 1920
atgagccgcg acgcgcaggg gatggtggaa gaagcgaagc gcctgcgcga cgctattccg 1980
ggtattgtgg tgaaaatccc ggtgacttcc gaaggtctgg cagcaattaa aatactgaaa 2040
aaagagggta ttactacact tggcactgct gtatatagcg ccgcacaagg gttattagcc 2100
gcactggcag gggcaaaata cgttgctccg tatgttaacc gcgtagatgc ccagggcgga 2160
gacggcattc gtacggttca ggagctgcaa acgctgttag aaatgcacgc gccagaaagc 2220
atggtgctgg cagccagctt taaaacgccg cgtcaggcgc tggactgttt actggcagga 2280
tgtgaatcca tcaccctgcc cttagatgta gcgcaacaaa tgctcaacac ccctgcggta 2340
gagtcagcta tagagaagtt cgaacacgac tggaatgccg catttggcac tactcatctc 2400
taaaggagca attatggacc gcattattca atcaccgggt aaatacatcc agggcgctga 2460
tgtgattaat cgtctgggcg aatacctgaa gccgctggca gaacgctggt tagtggtggg 2520
tgacaaattt gttttaggtt ttgctcaatc cactgtcgag aaaagcttta aagatgctgg 2580
actggtagta gaaattgcgc cgtttggcgg tgaatgttcg caaaatgaga tcgaccgtct 2640
gcgtggcatc gcggagactg cgcagtgtgg cgcaattctc ggtatcggtg gcggaaaaac 2700
cctcgatact gccaaagcac tggcacattt catgggtgtt ccggtagcga tcgcaccgac 2760
tatcgcctct accgatgcac cgtgcagcgc attgtctgtt atctacaccg atgagggtga 2820
gtttgaccgc tatctgctgt tgccaaataa cccgaatatg gtcattgtcg acaccaaaat 2880
cgtcgctggc gcacctgcac gtc 2903
<210>93
<211>79
<212>DNA
<213>人工序列
<220>
<223>atL-Ptac-fsaB
<400>93
tgcgtgccag acgttcgact tctgcgacgt tagcggtgtc cagatacagt tccatgatgt 60
cctgtgtgaa attgttatc 79
<210>94
<211>77
<212>DNA
<213>人工序列
<220>
<223>atR-Ptac-fsaB
<400>94
aacgccgcct ctgccgacgc tatcgccagc ctgctgcaac atgaactgga actgtaaatc 60
tagacgctca agttagt 77
Claims (7)
1.一种生产L-氨基酸的方法,包括在含有甘油作为碳源的培养基中培养具有L-氨基酸生产能力且经过修饰而增强了甘油脱氢酶和二羟基丙酮激酶的活性的大肠杆菌(Escherichia coli),从而在该培养基中或菌体内生成和蓄积L-氨基酸,并且从该培养基或菌体收集L-氨基酸,其中所述二羟基丙酮激酶以ATP作为磷酸供体。
2.权利要求1所述的方法,其中,所述酶的活性是通过提高编码所述甘油脱氢酶和二羟基丙酮激酶的基因的拷贝数和/或通过修饰所述基因的表达调节序列而增强的。
3.权利要求1所述的方法,其特征在于,所述二羟基丙酮激酶获自酿酒酵母(Saccharomyces cerevisiae)。
4.权利要求1所述的方法,其中,所述微生物还经过修饰而增强了甘油易化蛋白活性, 其中甘油易化蛋白活性是通过增加编码甘油易化蛋白的基因的拷贝数,和/或将所述基因的启动子更换成更强的启动子而增强的。
5.权利要求1所述的方法,其中,所述微生物还经过修饰而增强了选自由丙糖磷酸异构酶、果糖二磷酸醛缩酶、果糖1,6-二磷酸酶和果糖-6-磷酸醛缩酶组成的群组中的1种以上的酶活性。
6.权利要求1所述的方法,其中,所述微生物还经过修饰而降低了甘油激酶和/或膜结合型甘油-3-磷酸脱氢酶的酶活性。
7.权利要求1~6中任一项所述的方法,其中,所述L-氨基酸为选自由L-谷氨酸、L-赖氨酸、L-亮氨酸、L-异亮氨酸、L-缬氨酸、L-色氨酸、L-苯丙氨酸、L-酪氨酸、L-苏氨酸、L-甲硫氨酸、L-半胱氨酸、L-精氨酸、L-丝氨酸、L-脯氨酸、L-天冬氨酸、L-天冬酰胺、L-谷氨酰胺和L-组氨酸组成的群组中的1种或2种以上的L-氨基酸。
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US20130288313A1 (en) | 2013-10-31 |
WO2008102861A1 (ja) | 2008-08-28 |
BRPI0807756B1 (pt) | 2018-03-13 |
CN101631871A (zh) | 2010-01-20 |
RU2009135235A (ru) | 2011-03-27 |
RU2515044C2 (ru) | 2014-05-10 |
MX2009009025A (es) | 2009-09-03 |
CN103642863A (zh) | 2014-03-19 |
CA2689046A1 (en) | 2008-08-28 |
US20100047878A1 (en) | 2010-02-25 |
EP2113570A1 (en) | 2009-11-04 |
BRPI0807756A2 (pt) | 2014-06-17 |
EP2113570A4 (en) | 2013-11-13 |
US8512987B2 (en) | 2013-08-20 |
KR20100014538A (ko) | 2010-02-10 |
JP2010110217A (ja) | 2010-05-20 |
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