CN112368387A - 通过发酵产生组胺的经改造的生物合成途径 - Google Patents

通过发酵产生组胺的经改造的生物合成途径 Download PDF

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CN112368387A
CN112368387A CN201980036608.0A CN201980036608A CN112368387A CN 112368387 A CN112368387 A CN 112368387A CN 201980036608 A CN201980036608 A CN 201980036608A CN 112368387 A CN112368387 A CN 112368387A
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C·A·特雷西韦尔
A·G·希勒
M·S·西迪基
S·M·埃德加
N·赫尔曼
M·S·侯赛因
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Abstract

本公开内容描述了改造微生物细胞以用于组胺的发酵产生，并提供了新的经改造的微生物细胞和培养物，以及相关的组胺产生方法。

Description

通过发酵产生组胺的经改造的生物合成途径
相关申请的交叉引用
本申请要求2018年4月20日提交的美国临时申请号62/660,875的权益，该临时申请以引用的方式全文纳入本文。
关于在联邦资助的研究和开发下进行的发明的权利的声明
本发明在由DARPA授予的协议号为HR0011-15-9-0014下的政府支持下进行。政府对本发明拥有一定的权利。
以引用的方式纳入序列表
本申请包括序列表，该序列表以ASCII格式电子提交并且以引用的方式全文纳入本文。该ASCII副本于2019年4月17日生成，被命名为ZMGNP011WO_Seq_List_ST25.txt，大小为312,107字节。
技术领域
本公开内容总体上涉及改造微生物用于通过发酵产生组胺的领域。
背景技术
生物胺是具有生物活性的有机碱，其常存在于发酵食品和饮料中。已知组胺天然存在于发酵食品如酸奶(13-36mg/kg)[1]、味噌(24mg/kg)[2]和红酒(24mg/L)[3]中。一些生活在人肠道中的细菌也会产生组胺，并且它通过抗炎作用来起到调节免疫系统的功能[4]。发酵食品中组胺的产生依赖于含有组氨酸的蛋白质来源和组氨酸脱羧酶的微生物。组胺是组氨酸的脱羧产物，组氨酸由组氨酸脱羧酶(EC 4.1.1.22)特异性催化。从简单的非蛋白碳源和氮源在工业发酵中生产组胺需要装配具有氨基酸前体组氨酸和高活性组氨酸脱羧酶的改善的生物合成的途径。
发明内容
本公开内容提供了经改造的微生物细胞、微生物细胞的培养物以及用于产生组胺的方法，包括以下实施方案：
实施方案1：一种表达非天然的组氨酸脱羧酶的经改造的微生物细胞，其中所述经改造的微生物细胞产生组胺。
实施方案2：实施方案1的经改造的微生物细胞，其中所述经改造的微生物细胞包括一种或多种上游组胺途径酶的提高的活性，所述提高的活性为相对于对照细胞的提高。
实施方案3：实施方案2的经改造的微生物细胞，其中所述一种或多种上游组胺途径酶选自ATP磷酸核糖基转移酶、磷酸核糖基-ATP焦磷酸酶、磷酸核糖基-AMP环化水解酶、5’ProFAR异构酶、咪唑-甘油磷酸合酶、咪唑-甘油磷酸脱水酶、组氨醇-磷酸氨基转移酶、组氨醇-磷酸磷酸酶、组氨醇脱氢酶和磷酸核糖焦磷酸激酶。
实施方案4：实施方案1-3中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括一种或多种消耗一种或多种组胺途径前体的酶的降低的活性，所述降低的活性为相对于对照细胞的降低。
实施方案5：实施方案4的经改造的微生物细胞，其中所述一种或多种消耗一种或多种组胺途径前体的酶选自烯醇化酶、丙酮酸脱氢酶、戊糖磷酸途径糖异构酶、转醛醇酶、转酮醇酶、核酮糖-5-磷酸差向异构酶和核酮糖-5-磷酸异构酶。
实施方案6：实施方案4或实施方案5的经改造的微生物细胞，其中所述降低的活性是通过用活性较低的启动子替换所述一种或多种酶的基因的天然启动子来实现的。
实施方案7：实施方案1-6中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞额外地表达反馈失调的(feedback-deregulated)葡萄糖-6-磷酸脱氢酶或反馈失调的ATP磷酸核糖基转移酶。
实施方案8：一种经改造的微生物细胞，其中所述经改造的微生物细胞包括用于表达非天然的组氨酸脱羧酶的方式，其中所述经改造的微生物细胞产生组胺。
实施方案9：实施方案8的经改造的微生物细胞，其中所述经改造的微生物细胞包括用于提高一种或多种上游组胺途径酶的活性的方式，所述提高的活性为相对于对照细胞的提高。
实施方案10：实施方案9的经改造的微生物细胞，其中所述一种或多种上游组胺途径酶选自ATP磷酸核糖基转移酶、磷酸核糖基-ATP焦磷酸酶、磷酸核糖基-AMP环化水解酶、5’ProFAR异构酶、咪唑-甘油磷酸合酶、咪唑-甘油磷酸脱水酶、组氨醇-磷酸氨基转移酶、组氨醇-磷酸磷酸酶、组氨醇脱氢酶和磷酸核糖焦磷酸激酶。
实施方案11：实施方案8-10中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括用于降低一种或多种消耗一种或多种组胺途径前体的酶的活性的方式，所述降低的活性为相对于对照细胞的降低。
实施方案12：实施方案11的经改造的微生物细胞，其中所述一种或多种消耗一种或多种组胺途径前体的酶选自烯醇化酶、丙酮酸脱氢酶、戊糖磷酸途径糖异构酶、转酮醇酶、转醛醇酶、核酮糖-5-磷酸差向异构酶和核酮糖-5-磷酸异构酶。
实施方案13：实施方案11或实施方案12的经改造的微生物细胞，其中所述降低的活性是通过用活性较低的启动子替换所述一种或多种酶的基因的天然启动子的方式来实现的。
实施方案14：实施方案8-13中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞还包括用于表达葡萄糖-6-磷酸脱氢酶或反馈失调的ATP磷酸核糖基转移酶的方式。
实施方案15：实施方案1-14中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括真菌细胞。
实施方案16：实施方案15的经改造的微生物细胞，其中所述经改造的微生物细胞包括酵母细胞。
实施方案17：实施方案16的经改造的微生物细胞，其中所述酵母细胞为酵母属(Saccharomyces)或亚罗酵母属(Yarrowia)的细胞。
实施方案18：实施方案17的经改造的微生物细胞，其中所述酵母细胞为酵母属和酿酒(cerevisiae)种的细胞。
实施方案19：实施方案17的经改造的微生物细胞，其中所述酵母细胞为亚罗酵母属和解脂(lipolytica)种的细胞。
实施方案20：实施方案1-19中任一项的经改造的微生物细胞，其中所述非天然的组氨酸脱羧酶包括与来自色杆菌属种(Chromobacterium sp.)LK1或来自鲍氏不动杆菌(Acinetobacter baumannii)菌株AB0057的组氨酸脱羧酶具有至少70％氨基酸序列同一性的组氨酸脱羧酶。
实施方案21：实施方案1和16-20中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括一种或多种上游组胺途径酶的提高的活性，所述提高的活性为相对于对照细胞的提高，其中所述一种或多种上游组胺途径酶包括ATP磷酸核糖基转移酶。
实施方案22：实施方案21的经改造的微生物细胞，其中所述ATP磷酸核糖基转移酶的提高的活性是通过异源表达它来实现的。
实施方案23：实施方案22的经改造的微生物细胞，其中所述异源ATP磷酸核糖基转移酶与来自酿酒酵母(S.cerevisiae)的ATP磷酸核糖基转移酶具有至少70％氨基酸序列同一性。
实施方案24：实施方案16-23中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括谷氨酸棒状杆菌(Corynebacterium glutamicum)ATP磷酸核糖基转移酶的反馈失调的变体。
实施方案25：实施方案1-14中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞为细菌细胞。
实施方案26：实施方案25的经改造的微生物细胞，其中所述细菌细胞为棒状杆菌属(Corynebacteria)或芽孢杆菌属(Bacillus)的细胞。
实施方案27：实施方案26的经改造的微生物细胞，其中所述细菌细胞为棒状杆菌属和谷氨酸(glutamicum)种的细胞。
实施方案28：实施方案26的经改造的微生物细胞，其中所述细菌细胞为芽孢杆菌属和枯草(subtilis)种的细胞。
实施方案29：实施方案25-28中任一项的经改造的微生物细胞，其中所述非天然的组氨酸脱羧酶包括与来自鲍氏不动杆菌或来自乳杆菌属种(Lactobacillus sp.)(菌株30a)的组氨酸脱羧酶具有至少70％氨基酸序列同一性的组氨酸脱羧酶。
实施方案30：实施方案1和25-29中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括一种或多种上游组胺途径酶的提高的活性，所述提高的活性为相对于对照细胞的提高，其中所述一种或多种上游组胺途径酶包括ATP磷酸核糖基转移酶和咪唑-甘油磷酸脱水酶。
实施方案31：实施方案30的经改造的微生物细胞，其中所述ATP磷酸核糖基转移酶或咪唑-甘油磷酸脱水酶的提高的活性是通过异源表达它来实现的。
实施方案32：实施方案31的经改造的微生物细胞，其中所述异源ATP磷酸核糖基转移酶与来自酿酒酵母S288c或来自鼠伤寒沙门氏菌(Salmonella typhimurium)LT2的ATP磷酸核糖基转移酶具有至少70％氨基酸序列同一性或所述异源咪唑-甘油磷酸脱水酶与来自谷氨酸棒状杆菌的咪唑-甘油磷酸脱水酶具有至少70％氨基酸序列同一性。
实施方案33：实施方案25-32中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞包括鼠伤寒沙门氏菌ATP磷酸核糖基转移酶的反馈失调的变体。
实施方案34：实施方案1-33中任一项的经改造的微生物细胞，其中所述经改造的微生物细胞在培养时产生组胺，其水平为至少20mg/L培养基。
实施方案35：实施方案34的经改造的微生物细胞，其中所述经改造的微生物细胞在培养时产生组胺，其水平为至少300mg/L培养基。
实施方案36：实施方案1-35中任一项的经改造的微生物细胞的培养物。
实施方案37：实施方案36的培养物，其中所述经改造的微生物细胞以一定浓度存在，从而使得所述培养物在600nm下的光密度为10-500。
实施方案38：实施方案36-37中任一项的培养物，其中所述培养物包含组胺。
实施方案39：实施方案36-38中任一项的培养物，其中所述培养物包含水平至少为20mg/L培养基的组胺。
实施方案40：培养实施方案1-35中任一项的经改造的微生物细胞的方法，所述方法包括在适合产生组胺的条件下培养所述细胞。
实施方案41：实施方案40的方法，其中所述方法包括分批补料培养，初始葡萄糖水平范围为1-100g/L，然后是受控的糖补料。
实施方案42：实施方案40-41中任一项的方法，其中所述发酵底物包括葡萄糖和选自尿素、铵盐、氨及其任意组合的氮源。
实施方案43：实施方案40-42中任一项的方法，其中在培养过程中所述培养物是pH受控的。
实施方案44：实施方案40-43中任一项的方法，其中在培养过程中给所述培养物充气。
实施方案45：实施方案40-44中任一项的方法，其中所述经改造的微生物细胞以至少20mg/L培养基的水平产生组胺。
实施方案46：实施方案40-45中任一项的方法，其中所述方法还包括从培养物中回收组胺。
实施方案47：使用经改造以产生组胺的微生物细胞来制备组胺的方法，所述方法包括：(a)在微生物细胞中表达非天然的组氨酸脱羧酶；(b)在适合的培养基中，在使微生物细胞产生组胺的条件下培养微生物细胞，其中所述组胺被释放到培养基中；和从培养基中分离组胺。
附图说明
图1：组胺的生物合成途径。
图2：第一轮经改造的宿主谷氨酸棒状杆菌发酵之后，在细胞外培养液中测定的组胺效价。(另见实施例1，表1)
图3：第一轮经改造的宿主酿酒酵母发酵之后，在细胞外培养液中测定的组胺效价。(另见实施例1，表1)
图4：第二轮经改造的宿主谷氨酸棒状杆菌发酵之后，在细胞外培养液中测定的组胺效价。(另见实施例1，表2)
图5：第二轮经改造的宿主酿酒酵母发酵之后，在细胞外培养液中测定的组胺效价。(另见实施例1，表2)
图6：第一轮经改造的宿主解脂亚罗酵母发酵之后，在细胞外培养液中测定的组胺效价。(另见实施例2，表4)
图7：第一轮经改造的宿主枯草芽胞杆菌发酵之后，在细胞外培养液中测定的组胺效价。
图8：表达宿主评估设计的酿酒酵母发酵之后，在细胞外培养液中测定的组胺酸效价。
图9：表达宿主评估设计的谷氨酸棒状杆菌发酵之后，在细胞外培养液中测定的组胺酸效价。
图10：第三轮经改造的宿主酿酒酵母发酵之后，在细胞外培养液中测定的组胺效价。(提高轮)
图11：将启动子-基因-终止子整合到酿酒酵母和解脂亚罗酵母中。
图12：酿酒酵母和解脂亚罗酵母中的启动子替换。
图13：酿酒酵母和解脂亚罗酵母中的靶基因缺失。
图14：将启动子-基因-终止子整合到谷氨酸棒状杆菌和枯草芽胞杆菌中。
具体实施方式
本公开内容描述了由微生物宿主分别从简单的碳源和氮源(如葡萄糖和尿素)通过发酵产生小分子组胺的方法。这一目的可以通过将非天然的代谢途径引入到适合的微生物宿主中以用于大规模化学产品的工业发酵来实现。示例性宿主包括酿酒酵母、解脂亚罗酵母、谷氨酸棒状杆菌和枯草芽孢杆菌。经改造的代谢途径将宿主的中心代谢与非天然的途径联系起来，从而能够产生组胺。这种方法最简单的实施方案是在能产生组氨酸的微生物宿主菌株中表达一种酶，即非天然的组氨酸脱羧酶。通过关键的上游途径酶ATP磷酸核糖基转移酶的过表达和突变来修饰微生物宿主中心代谢，进一步改造代谢途径，能够使效价达到505mg/L组胺。
以下公开内容描述了如何改造具有必要特性的微生物，以从简单的碳源和氮源产生工业上可行的组胺效价。已鉴定了活性组氨酸脱羧酶，并且已发现反馈失调的ATP磷酸核糖基转移酶和/或天然ATP磷酸核糖基转移酶的组成型表达通过发酵提高了组氨酸的效价。
定义
除非另外具体说明，权利要求书和说明书中使用的术语如下文所定义。
术语“发酵”在本文中用于指这样的过程，即其中微生物细胞通过一个或多个生物转化步骤将一种或多种底物转化为所需产物(例如组胺)，而不需要任何化学转化步骤。
本文中使用的术语“经改造的(engineered)”与细胞有关，用于表示所述细胞包含至少一种人工引入的靶向遗传改变，该遗传改变使经改造的细胞区别于天然存在的细胞。
术语“天然的”在本文中用于指天然存在于特定细胞中的细胞组分，例如多核苷酸或多肽。天然多核苷酸或多肽是细胞内源性的。
当与多核苷酸或多肽有关时，术语“非天然的”是指非天然存在于特定细胞中的多核苷酸或多肽。
当与基因表达的环境有关时，术语“非天然的”是指在其天然表达的基因组和细胞环境之外的任何环境中表达的基因。以非天然方式表达的基因可能与宿主细胞中相应基因具有相同的核苷酸序列，但可能从载体表达或从基因组中与天然基因的基因座不同的整合点表达。
术语“异源的”在本文中用于描述被引入到宿主细胞中的多核苷酸或多肽。该术语包括分别来自不同于宿主细胞的生物体、物种或菌株的多核苷酸或多肽。在这种情况下，异源的多核苷酸或多肽的序列不同于同一宿主细胞中存在的任何序列。然而，该术语还包括其序列与在宿主细胞中存在的序列相同的多核苷酸或多肽，其中所述多核苷酸或多肽存在于与天然序列不同的环境中(例如，异源多核苷酸可以连接到与天然序列不同的启动子上，并插入到不同的基因组位置)。因此，“异源表达”包括宿主细胞非天然序列的表达，以及在非天然环境下宿主细胞天然序列的表达。
如关于多核苷酸或多肽所使用，术语“野生型”是指具有存在于来自天然存在的生物体的多核苷酸或多肽中的核苷酸序列的任何多核苷酸或具有存在于来自天然存在的生物体的多核苷酸或多肽中的氨基酸序列的任何多肽，无论分子来源如何；即，术语“野生型”是指序列特征，无论所述分子是从天然来源中纯化；重组表达，然后纯化；还是合成的。术语“野生型”还用于指天然存在的细胞。
“对照细胞”是这样的细胞，其在其他方面与测试的经改造的细胞相同，包括与经改造的细胞为相同属和种，但是缺少在经改造的细胞中测试的特定遗传修饰。
在本文中，酶是通过它们催化的反应来鉴定，除非另外说明，酶是指能够催化所鉴定的反应的任何多肽。除非另外说明，酶可源自任何生物体，并且可具有天然的或突变的氨基酸序列。本领域中公知，酶可以具有多种功能和/或多个名称，有时取决于它们所源自的生物体。本文使用的酶名称涵盖了直系同源物，包括可具有一种或多种额外的功能或不同名称的酶。
本文中使用的术语“反馈失调的”与在特定细胞中通常被酶通路的下游产物负调控(即反馈抑制)的酶有关。在这种情况下，“反馈失调的”酶是这种形式的酶，其与细胞原生的天然酶相比对反馈抑制更加不敏感。反馈失调的酶可以通过将一种或多种突变引入到天然酶中而产生。或者，反馈失调的酶可以仅仅为异源天然酶，当其被引入到特定微生物细胞中时，对反馈抑制的敏感度不如原生的天然酶。在一些实施方案中，反馈失调的酶在微生物细胞中不显示出反馈抑制。
术语“组胺”是指2-(1I-咪唑-4-基)乙胺(CAS#51-45-6)。
在两个或多个氨基酸或核苷酸序列的情况下，术语“序列同一性”是指当进行比较并最大对应性的比对时，相同或具有特定的相同氨基酸残基或核苷酸百分比的两个或多个序列，如使用序列比较算法或通过目测来测量的。
对于序列比较以确定百分比核苷酸或氨基酸序列同一性，通常一个序列作为“参考序列”，将“测试”序列与之进行比较。当使用序列比较算法时，将测试序列和参考序列输入到计算机中，如果必要，指定子序列坐标，并且指定序列算法程序参数。然后基于指定的程序参数，序列比较算法计算测试序列相对于参考序列的百分比序列同一性。使用设置默认参数的BLAST，可以进行序列比对以进行比较。
本文使用的术语“效价”是指由微生物细胞培养物产生的产物(例如组胺)的质量除以培养物体积。
关于从细胞培养物中回收组胺，本文使用的“回收”是指从至少一种细胞培养基的其他组分中分离出组胺。
改造微生物用于组胺产生
组胺生物合成途径
组胺通常源自氨基酸组氨酸。组胺生物合成途径如图1所示。氨基酸生物合成途径的第一种酶——ATP磷酸核糖基转移酶受到组氨酸的反馈抑制。通过在酿酒酵母、解脂亚罗酵母、谷氨酸棒状杆菌和枯草芽孢杆菌宿主中添加单一的非天然的酶促步骤实现组胺产生，该酶促步骤由组氨酸脱羧酶(EC 4.1.1.22)催化。
用于微生物组胺产生的改造
可将在被改造的微生物细胞中具备活性的任何组氨酸脱羧酶引入到细胞中，通常通过使用标准的基因工程技术引入和表达编码所述酶的基因。适合的组氨酸脱羧酶可来自任何来源，包括植物、古细菌、真菌、革兰氏阳性细菌和革兰氏阴性细菌来源。示例性来源包括但不限于：杀鲑气单胞菌溶果胶亚种(Aeromonas salmonicida subsp.pectinolytica)34mel、鲍氏不动杆菌(菌株AB0057)、溶血色杆菌(Chromobacterium haemolyticum)、色杆菌属种LK1、巴氏柠檬酸杆菌(Citrobacter pasteurii)、黑腹果蝇(Drosophilamelanogaster)、鸟类乳酸菌(Lactobacillus aviaries)DSM 20655、食果糖乳杆菌(Lactobacillus fructivorans)、罗伊乳杆菌(Lactobacillus reuteri)、乳杆菌属种(菌株30a)、巴氏甲烷八叠球菌(Methanosarcina barkeri)(菌株Fusaro/DSM804)、巴氏甲烷八叠球菌菌株Wiesmoor、耐冷摩根氏菌(Morganella psychrotolerans)、小家鼠(Musmusculus)、酒类酒球菌(Oenococcus oeni)(酒明串珠菌(Leuconostoc oenos))、恶臭假单胞菌(Pseudomonas putida)(铁荚膜节杆菌(Arthrobacter siderocapsulatus))、根际假单胞菌(Pseudomonas rhizosphaerae)、假单胞菌属种(Pseudomonas sp.)bs2935、番茄(Solanum lycopersicum)、稻(Oryza sativa)、马尔尼菲青霉菌(Penicilliummarneffei)、吸水链霉菌(Streptomyces hygroscopicus)、恶臭假单胞菌、拟南芥(Arabidopsis thaliana)(水芹鼠耳草(Mouse-ear cress))、野生大豆(Glycine soja)(野生大豆(Wild soybean))、番茄(番茄(Tomato))(番茄(Lycopersicon esculentum))、产气荚膜梭菌(Clostridium perfringens)、布赫内氏乳杆菌(Lactobacillus buchneri)、黑腹果蝇(果蝇(Fruit fly))、摩氏摩根菌(Morganella morganii)(摩氏变形杆菌(Proteusmorganii))、大肠杆菌、牛(Bos taurus)(牛(Bovine))、Raoutella planticol(植生克雷伯氏菌(Klebsiella planticola))、鲍氏不动杆菌、溶血不动杆菌(Acinetobacterhaemolyticus)、美人鱼发光杆菌(Photobacterium damselae)、盐水四联球菌(Tetragenococcus muriaticus)、南极细菌属种(Moritellasp)JT01、嗜热链球菌(Streptococcus thermophilus)、产气肠杆菌(Enterobacter aerogenes)、杨氏柠檬酸杆菌(Citrobacter youngae)、解鸟氨酸拉乌尔菌(Raoultella omithinolytica)和植生拉乌尔菌(Raoultella planticola)。
可将组氨酸脱羧酶基因的一个或多个拷贝引入选择的微生物宿主细胞。如果引入多于一个拷贝的基因，则所述拷贝可具有相同或不同核苷酸序列。在一些实施方案中，从强的组成型启动子表达一种或两种异源基因。在一些实施方案中，从诱导型启动子表达异源的组氨酸脱羧酶基因。异源基因可以任选地进行密码子优化，以增强在选择的微生物宿主细胞中的表达。实施例中使用的宿主的示例性密码子优化表如下：枯草芽孢杆菌Kazusa密码子表：www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi？species＝1423&aa＝1&style＝N；解脂亚罗酵母Kazusa密码子表：www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi？species＝4952&aa＝1&style＝N；谷氨酸棒状杆菌Kazusa密码子表：www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi？species＝340322&aa＝1&style＝N；酿酒酵母Kazusa密码子表：http：//www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi？species＝4932&aa＝1&style＝N。还使用了用于酿酒酵母和谷氨酸棒状杆菌的改良组合密码子使用方案，其在下文中再次出现。
改良的Sc和Cg的密码子使用表
Figure BDA0002807770440000111
Figure BDA0002807770440000121
提高上游酶的活性
在能够产生组胺的微生物细胞中提高组胺产生的一个方法是提高组胺生物合成途径中的一种或多种上游酶的活性。上游途径酶包括从进料直到最终天然代谢物(在以下实施例中描述的示例性微生物细胞中的组氨酸)的转化中涉及的所有酶。这样的酶包括ATP磷酸核糖基转移酶、磷酸核糖基-ATP焦磷酸酶、磷酸核糖基-AMP环化水解酶、5’ProFAR异构酶、咪唑-甘油磷酸合酶、咪唑-甘油磷酸脱水酶、组氨醇-磷酸氨基转移酶、组氨醇-磷酸磷酸酶、组氨醇脱氢酶和磷酸核糖焦磷酸激酶。适合的编码这些酶的上游途径基因可来自任何来源，包括例如，上文中作为组氨酸脱羧酶基因的来源被讨论的那些。
在一些实施方案中，通过调节天然酶的表达或活性来提高一种或多种上游途径酶的活性。例如，可以利用这些酶的表达或活性的天然调节因子来提高适合的酶的活性。
或者，或另外，使用例如图12所示的技术，一个或多个启动子可以置换天然启动子。在某些实施方案中，替换启动子比天然启动子更强，和/或为组成型启动子。
在一些实施方案中，一种或多种上游途径酶的活性通过将一种或多种相应的基因引入到表达组氨酸脱羧酶的微生物宿主细胞中来补充。所引入的上游途径基因可能来自宿主细胞以外的生物体，或仅为天然基因的额外拷贝。在一些实施方案中，将一种或多种这样的基因引入到能够产生组胺的微生物宿主细胞中，并从强的组成型启动子表达，和/或可任选地对所述基因进行密码子优化以增强在选择的微生物宿主细胞中的表达。
实施例1描述了成功改造谷氨酸棒状杆菌以表达来自鲍氏不动杆菌的异源组胺脱羧酶(SEQ ID NO：1)，以及组成型表达异源谷氨酸棒状杆菌咪唑甘油-磷酸脱水酶(SEQ IDNO：2)。该菌株产生自两轮基因改造并以24mg/L培养基的效价产生组胺。该效价在经改造以表达来自鲍氏不动杆菌(菌株AB0057)的组胺脱羧酶(SEQ ID NO：1)和来自酿酒酵母S288c的ATP磷酸核糖基转移酶(SEQ ID NO：3)的谷氨酸棒状杆菌中提高到68mg/L。
实施例2描述了成功改造解脂亚罗酵母以表达来自鲍氏不动杆菌(菌株AB0057)的组氨酸脱羧酶(SEQ ID NO：1)和来自酿酒酵母S288c的ATP磷酸核糖基转移酶(SEQ ID NO：3)，以得到505mg/L的组胺效价。实施例2还描述了改造枯草芽孢杆菌以表达来自乳杆菌属种(菌株30a)的组胺脱羧酶(SEQ ID NO：4)和来自鼠伤寒沙门氏菌LT2的ATP磷酸核糖基转移酶(SEQ ID NO：5)，以得到18mg/L的组胺效价。还在实施例2中改造酿酒酵母以表达来自色杆菌属种LK1的组胺脱羧酶(SEQ ID NO：6)和来自酿酒酵母S288c的ATP磷酸核糖基转移酶(SEQ ID NO：3)，以得到111mg/L的组胺效价。
在各实施方案中，改造产生组胺的微生物细胞以提高一种或多种上游途径酶的活性，这将组胺效价提高至少10％、20％、30％、40％、50％、60％、70％、80％或90％，或至少2倍、2.5倍、3倍、3.5倍、4倍、4.5倍、5倍、5.5倍、6倍、6.5倍、7倍、7.5倍、8倍、8.5倍、9倍、9.5倍、10倍、11倍、12倍、13倍、14倍、15倍、16倍、17倍、18倍、19倍、20倍、21倍、22倍、23倍、24倍、25倍、30倍、35倍、40倍、45倍、50倍、55倍、60倍、65倍、70倍、75倍、80倍、85倍、90倍、95倍或100倍。在各实施方案中，组胺效价的提高在10％至100倍、2倍至50倍、5倍至40倍、10倍至30倍的范围内，或以上列出的任何数值界定的任何范围内。(本文的范围包括它们的端点。)这些提高相对于在上游途径酶活性没有任何提高的产组胺微生物细胞中测定的组胺效价而确定。这种参考细胞可具有一种或多种旨在增加组胺产生的其他遗传改变，例如，所述细胞可表达反馈失调的酶。
在各实施方案中，通过提高一种或多种上游途径基因的活性而实现的组胺效价为至少1、10、20、30、40、50、75、100、200、300、400、500、600、700、800或900mg/L或至少1、1.5、2、2.5、3、3.5、4、4.5、5或10gm/L。在各实施方案中，所述效价在10mg/L至10gm/L、20mg/L至5gm/L、50mg/L至4gm/L、100mg/L至3gm/L、500mg/L至2gm/L的范围内，或以上列出的任何数值界定的任何范围内。
反馈失调的酶的引入
由于组氨酸生物合成受到反馈抑制，在经改造以产生组胺的微生物细胞中提高组胺产生的另一种方法是引入通常受到反馈调节的一种或多种酶的反馈失调形式。这些酶的实例包括葡萄糖-6-磷酸脱氢酶和ATP磷酸核糖基转移酶。反馈失调形式可以是异源、天然酶，其与特定微生物宿主细胞中的天然酶相比对反馈抑制更不敏感。或者，反馈失调形式可以是天然或异源酶的变体，其具有一个或多个突变或被截短，使其与对应的天然酶相比对反馈抑制更不敏感。后者的实例包括含有氨基酸置换N215K、L231F和T235A的变体ATP磷酸核糖基转移酶(来自谷氨酸棒状杆菌)(SEQ ID NO：7)和含有氨基酸Q207和E208缺失的变体ATP磷酸核糖基转移酶(来自鼠伤寒沙门氏菌)(SEQ ID NO：5)。
在各实施方案中，改造产生组胺的微生物细胞以表达反馈失调的酶，这将组胺的效价提高至少10％、20％、30％、40％、50％、60％、70％、80％或90％，或至少2倍、2.5倍、3倍、3.5倍、4倍、4.5倍、5倍、5.5倍、6倍、6.5倍、7倍、7.5倍、8倍、8.5倍、9倍、9.5倍、10倍、11倍、12倍、13倍、14倍、15倍、16倍、17倍、18倍、19倍、20倍、21倍、22倍、23倍、24倍、25倍、30倍、35倍、40倍、45倍、50倍、55倍、60倍、65倍、70倍、75倍、80倍、85倍、90倍、95倍或100倍。在各实施方案中，组胺效价的提高在10％至100倍、2倍至50倍、5倍至40倍、10倍至30倍的范围内，或以上列出的任何数值界定的任何范围内。这些提高相对于在不表达反馈失调的酶的产组胺微生物细胞中测定的组胺效价而确定。这种参考细胞可以(但是不需要)具有旨在增加组胺产生的其他遗传改变，即所述细胞可具有上游途径酶的提高的活性，这是由反馈不敏感以外的一些方式引起的。
在各实施方案中，通过使用反馈失调的酶以增加经过组胺生物合成途径的通量，实现的组胺效价为至少100、200、300、400、500、600、700、800或900μg/L，或至少1、10、50、75、100、200、300、400、500、600、700、800或900mg/L或至少1、1.5、2、2.5、3、3.5、4、4.5、5、10、20、50g/L。在各实施方案中，效价在50μg/L至50g/L、75μg/L至20g/L、100μg/L至10g/L、200μg/L至5g/L、500μg/L至4g/L、1mg/L至3g/L、500mg/L至2g/L的范围内，或以上列出的任何数值界定的任何范围内。
补充一种或多种天然酶的活性和/或引入一种或多种反馈失调的酶的方法可以在表达组胺脱羧酶的微生物细胞中结合以实现甚至更高的组胺产生水平。例如，酿酒酵母在两轮改造中通过引入三个基因，获得了385mg/L的组胺效价，这三个基因为：组氨酸脱羧酶基因(来自色杆菌属种LK1)(SEQ ID NO：6)、含有氨基酸置换N215K、L231F和T235A的ATP磷酸核糖基转移酶(来自谷氨酸棒状杆菌)(SEQ ID NO：7)和来自酿酒酵母S288c的组成型表达的ATP磷酸核糖基转移酶(SEQ ID NO：3)。(实施例1。)
减少前体消耗
在能够产生组胺的微生物细胞中增加组胺产生的另一种方法是降低一种或多种消耗一种或多种组胺途径前体的酶的活性。在一些实施方案中，通过调节天然酶的表达或活性来降低一种或多种这样的酶的活性。该类型的示例性酶包括烯醇化酶、丙酮酸脱氢酶、磷酸戊糖途径糖异构酶、转醛醇酶、转酮醇酶、核酮糖-5-磷酸差向异构酶和核酮糖-5-磷酸异构酶(aribulose-5-phosphate isomerase)。例如，用活性较低或失活的启动子替换对应基因的天然启动子，或者通过缺失对应的基因，可以使这样的酶的活性降低。参见图12和图13，其分别为酿酒酵母和解脂亚罗酵母中启动子替换和靶基因缺失的方案的实例。
在各实施方案中，改造产生组胺的微生物细胞以减少一个或多个旁路对前体的消耗，这将组胺效价提高至少10％、20％、30％、40％、50％、60％、70％、80％或90％，或至少2倍、2.5倍、3倍、3.5倍、4倍、4.5倍、5倍、5.5倍、6倍、6.5倍、7倍、7.5倍、8倍、8.5倍、9倍、9.5倍、10倍、11倍、12倍、13倍、14倍、15倍、16倍、17倍、18倍、19倍、20倍、21倍、22倍、23倍、24倍、25倍、30倍、35倍、40倍、45倍、50倍、55倍、60倍、65倍、70倍、75倍、80倍、85倍、90倍、95倍或100倍。在各实施方案中，组胺效价的提高在10％至100倍、2倍至50倍、5倍至40倍、10倍至30倍的范围内，或以上列出的任何数值界定的任何范围内。这些提高相对于在不包括减少前体消耗的遗传改变的产组胺微生物细胞中测定的组胺效价而确定。这种参考细胞可以(但是不需要)具有旨在增加组胺产生的其他遗传改变，即所述细胞可具有上游途径酶的提高的活性。
在各实施方案中，通过减少一个或多个旁路对前体的消耗，实现的组胺效价为至少100、200、300、400、500、600、700、800或900μg/L，或至少1、10、50、75、100、200、300、400、500、600、700、800或900mg/L或至少1、1.5、2、2.5、3、3.5、4、4.5、5、10、20、50g/L。在各实施方案中，效价在50μg/L至50g/L、75μg/L至20g/L、100μg/L至10g/L、200μg/L至5g/L、500μg/L至4g/L、1mg/L至3g/L、500mg/L至2g/L的范围内，或以上列出的任何数值界定的任何范围内。
增加一种或多种天然酶的活性和/或引入一种或多种反馈失调的酶和/或减少一个或多个旁路对前体消耗的方法可以结合以实现甚至更高的组胺产生水平。
微生物宿主细胞
可以改造任何可用于表达引入基因的微生物，用于如上文所述发酵产生组胺。在某些实施方案中，微生物是天然无法发酵产生组胺的微生物。在一些实施方案中，微生物是易于培养的微生物，例如，已知适用于作为目的化合物发酵产生中的宿主细胞的微生物。可以如上文所述，改造细菌细胞，包括革兰氏阳性细菌或革兰氏阴性细菌。除谷氨酸棒状杆菌细胞以外，实例还包括枯草芽孢杆菌、地衣芽孢杆菌(B.licheniformis)、迟缓芽孢杆菌(B.lentus)、短芽孢杆菌(B.brevis)、嗜热脂肪芽孢杆菌(B.stearothermophilus)、嗜碱芽孢杆菌(B.alkalophilus)、解淀粉芽孢杆菌(B.amyloliquefaciens)、克劳氏芽孢杆菌(B.clausii)、耐盐芽孢杆菌(B.halodurans)、巨大芽孢杆菌(B.megaterium)、凝结芽孢杆菌(B.coagulans)、环状芽孢杆菌(B.circulans)、灿烂芽孢杆菌(B.lautus)、苏云金芽孢杆菌(B.thuringiensis)、白色链霉菌(S.albus)、变铅青链霉菌(S.lividans)、天蓝色链霉菌(S.coelicolor)、灰色链霉菌(S.griseus)、假单胞菌属种、产碱假单胞菌(P.alcaligenes)、柑橘假单胞菌(P.citrea)、乳杆菌属种(Lactobacilis spp.)(例如乳酸乳杆菌(L.lactis)、植物乳杆菌(L.plantarum))、灰色乳杆菌(L.grayi)、大肠杆菌、粪肠球菌(E.faecium)、鹑鸡肠球菌(E.gallinarum)、酪黄肠球菌(E.casseliflavus)、和/或粪肠球菌(E.faecalis)细胞。
有多种类型的厌氧细胞可用作本文所述方法中的微生物宿主细胞。在一些实施方案中，微生物细胞是专性厌氧细胞。专性厌氧菌在存在氧的条件下通常无法良好生长(如果不是完全不生长的话)。应理解，可存在少量氧，即专性厌氧菌对低水平的氧具有一定的耐受水平。如上文所述改造的专性厌氧菌可生长在基本无氧的条件下，其中存在的氧的量对厌氧菌的生长、维持和/或发酵无害。
或者，用于本文所述方法的微生物宿主细胞可以是兼性厌氧细胞。如果存在氧，兼性厌氧菌可通过有氧呼吸(例如利用TCA循环)产生细胞ATP。但是，兼性厌氧菌还可在无氧情况下生长。如上文所述改造的兼性厌氧菌可以在基本无氧的条件下生长，其中存在的氧的量对厌氧菌的生长、维持和/或发酵无害，或者可以在存在较大量的氧的情况下生长。
在一些实施方案中，用于本文所述方法的微生物宿主细胞是丝状真菌细胞。(参见，例如Berka&Barnett，Biotechnology Advances，(1989)，7(2)：127-154)。实例包括长枝木霉(Trichodermalongibrachiatum)、绿色木霉(T.viride)、康氏木霉(T.koningii)、哈茨木霉(T.harzianum)、青霉菌属种(Penicillium sp.)、特异腐质霉(Humicola insolens)、H.lanuginose、灰腐质霉(H.grisea)、金孢子菌属种(Chrysosporium sp.)、鲁克文金孢菌(C.lucknowense)、粘帚霉属种(Gliocladium sp.)、曲霉属种(Aspergillus sp.)(例如，米曲霉(A.oryzae)、黑曲霉(A.niger)、酱油曲霉(A.sojae)、日本曲霉(A.japonicus)、构巢曲霉(A.nidulans)或泡盛曲霉(A.awamori))、镰刀菌属种(Fusarium sp.)(例如，粉红镰刀菌(F.roseum)、禾谷镰刀菌(F.graminum)、禾谷镰刀菌(F.cerealis)、F.oxysporuim或镶片镰孢霉(F.venenatum))、链孢霉属种(Neurospora sp.)(例如，粗糙链孢霉(N.crassa)或肉座菌属种(Hypocrea sp.))、毛霉属种(Mucor sp.)(例如，白色毛霉(M.miehei))、根霉属种(Rhizopus sp.)和裸胞壳属种(Emericella sp.)细胞。在具体实施方案中，如上文所述被改造的真菌细胞是构巢曲霉、泡盛曲霉、米曲霉、棘孢曲霉(A.aculeatus)、黑曲霉、日本曲霉、里氏木霉(T.reesei)、绿色木霉、F.oxysporum或腐皮镰刀菌(F.solani)。用于这样的宿主的示例性质粒或质粒组件包括美国专利公开号2011/0045563中描述的那些。
酵母还可用作本文所述方法中的微生物宿主细胞。实例包括：酵母属种(Saccharomyces sp.)、裂殖酵母属种(Schizosaccharomyces sp.)、毕赤酵母属种(Pichiasp.)、多形汉逊酵母(Hansenula polymorpha)、叶柄毕赤酵母(Pichia stipites)、马克斯克鲁维酵母(Kluyveromyces marxianus)、克鲁维酵母属种(Kluyveromyces spp.)、解脂亚罗酵母和假丝酵母属种(Candida sp.)。在一些实施方案中，酵母属种是酿酒酵母(参见，例如Romanos et al.，Yeast，(1992)，8(6)：423-488)。用于这类宿主的示例性质粒或质粒组件包括美国专利号7,659,097和美国专利公开号2011/0045563中描述的那些。
在一些实施方案中，宿主细胞可以是藻类细胞，其来自绿藻、红藻、灰胞藻(glaucophyte)、绿蜘藻(chlorarachniophyte)、眼虫(euglenid)、假菌界(chromista)或鞭毛藻(dinoflagellate)。(参见例如，Saunders&Warmbrodt，“Gene Expression in Algaeand Fungi，Including Yeast，”(1993)，National Agricultural Library，Beltsville，Md.)。用于藻类细胞的示例性质粒或质粒组件包括美国专利公开号2011/0045563中描述的那些。
在其他实施方案中，宿主细胞是蓝细菌，例如根据形态学归类到以下组中任一组的蓝细菌：绿球藻目(Chlorococcales)、宽球藻目(Pleurocapsales)、颤藻目(Oscillatoriales)、念珠藻目(Nostocales)、Synechosystic或真枝藻目(Stigonematales)(参见，例如Lindberg et al.，Metab.Eng.，(2010)12(1)：70-79)。用于蓝细菌细胞的示例性质粒或质粒组件包括美国专利公开号2010/0297749和2009/0282545以及国际专利公开号WO 2011/034863中描述的那些。
基因工程方法
使用本领域技术范围内的常规的分子生物学(包括重组技术)、微生物学、细胞生物学和生物化学技术，可以改造微生物细胞，用于发酵产生组胺。这类技术在文献中充分解释，参见，例如“Molecular Cloning：A Laboratory Manual，”fourth edition(Sambrooket al.，2012)；“Oligonucleotide Synthesis”(M.J.Gait，ed.，1984)；“Culture ofAnimal Cells：A Manual of Basic Technique and Specialized Applications”(R.I.Freshney，ed.，6th Edition，2010)；“Methods in Enzymology”(Academic Press，Inc.)；“Current Protocols in Molecular Biology”(F.M.Ausubel et al.，eds.，1987，and periodic updates)；“PCR：The Polymerase Chain Reaction，”(Mullis et al.，eds.，1994)；Singleton et al.，Dictionary of Microbiology and Molecular Biology2nd ed.，J.Wiley&Sons(New York，N.Y.1994)。
载体是用于将遗传物质引入到细胞中的多核苷酸载体。适用于本文所述方法的载体可以是线性的或环状的。载体可整合到宿主细胞的靶基因组中或在宿主细胞中独立复制。对于许多应用而言，优选产生稳定转化体的整合载体。载体可包含，例如，复制起点、多克隆位点(MCS)和/或选择性标记。表达载体通常包括含有调控元件的表达盒，所述调控元件促使多核苷酸序列(通常为编码序列)在特定宿主细胞中的表达。载体包括但不限于，整合载体、原核质粒、附加体(episome)、病毒载体、粘粒和人工染色体。
可用于表达盒的示例性调控元件包括启动子、增强子、内部核糖体进入位点(IRES)和其他表达调控元件(例如转录终止信号，如多腺苷酸化信号和poly-U序列)。这类调控元件记载于，例如Goeddel，Gene Expression Technology：Methods In Enzymology185，Academic Press，San Diego，Calif.(1990)。
在一些实施方案中，载体可用于引入能够进行基因组编辑的系统，例如CRISPR系统。参见2014年3月6日公布的美国专利公开号2014/0068797；还可参见Jinek M.，et al.，“A programmable dual-RNA-guided DNA endonuclease in adaptive bacterialimmunity，”Science 337：816-21，2012)。在II型CRISPR-Cas9系统中，Cas9是定点核酸内切酶，即使用两个不同的核酸内切酶结构域(HNH和RuvC/类RNase H结构域)可被定向以在特定靶序列切割多核苷酸的酶。可改造Cas9以在任何所需位点切割DNA，因为Cas 9是由RNA定向到其切割位点。因此，Cas9还被描述为“RNA引导的核酸酶”。更具体而言，Cas9与一个或多个RNA分子连接，所述RNA分子将Cas9导向至特定的靶多核苷酸，这是基于所述RNA分子的至少一部分与靶多核苷酸的特定序列杂交。Ran，F.A.等人(“知vivo genome editing usingStaphylococcus aureus Cas9，”Nature 520(7546)：186-91，2015，Apr 9]，包括全部扩展数据)提供了crRNA/tracrRNA序列和八种II型CRISPR-Cas9系统的二级结构。类Cas9合成蛋白也是本领域已知的(参见2014年10月23日公开的美国公开专利申请号2014-0315985)。
实施例1描述了用于将多核苷酸和其他遗传改变引入到谷氨酸棒状杆菌和酿酒酵母细胞的基因组中的示例性整合方法。
通过多种标准方法的任一种可将载体或其他多核苷酸引入到微生物细胞中，所述方法例如转化、接合、电穿孔、核微注射、转导、转染(例如，脂质转染介导的或DEAE-糊精介导的转染或使用重组噬菌体病毒的转染)、使用磷酸钙DNA沉淀物孵育、使用DNA包被的微粒高速轰击和原生质体融合。通过本领域已知的任何方法可选择转化体。适用于选择转化体的方法记载于美国专利公开号2009/0203102、2010/0048964和2010/0003716，以及国际公开号WO 2009/076676、WO 2010/003007和WO 2009/132220。
经改造的微生物细胞
上述方法可用于产生经改造的微生物细胞，在某些实施方案中，所述经改造的微生物细胞过量产生组胺。与天然微生物细胞(例如本文所述的任何微生物宿主细胞)相比，经改造的微生物细胞可具有至少1、2、3、4、5、6、7、8、9、10、20、30、40、50、60、70、80、90、100或更多个遗传改变，例如30-100个改变。下文实施例中描述的经改造的微生物细胞具有1、2或3个遗传改变，但是本领域技术人员按照本文的指导能够设计具有其他改变的微生物细胞。在一些实施方案中，与天然微生物细胞相比，经改造的微生物细胞具有不多于15、14、13、12、11、10、9、8、7、6、5或4个遗传改变。在各实施方案中，经改造以产生组胺的微生物细胞可具有落入以下任何示例性范围的多个遗传改变：1-10、1-9、1-8、2-7、2-6、2-5、2-4、2-3、3-7、3-6、3-5、3-4等。
在一些实施方案中，经改造的微生物细胞表达至少一种异源的组胺脱羧酶，如在无法天然产生组胺的微生物宿主细胞的情况下。在各实施方案中，微生物细胞可包含并表达，例如：(1)单一异源组胺脱羧酶基因，(2)两种或多种异源组胺脱羧酶基因，其可相同或不同(换言之，可引入相同的异源组胺脱羧酶基因的多个拷贝，或者可引入多种不同的异源组胺脱羧酶基因)，(3)不是细胞原生的单一异源组胺脱羧酶基因以及天然组胺脱羧酶基因的一个或多个额外拷贝，或(4)两种或多种非天然的组胺脱羧酶基因，其可相同或不同，以及天然组胺脱羧酶基因的一个或多个额外拷贝。
这种经改造的宿主细胞可包含至少一种其他遗传改变，其增加经由所述途径产生组氨酸(组胺的直接前体)的通量。途径中的这些“上游”酶包括：ATP磷酸核糖基转移酶、磷酸核糖基-ATP焦磷酸酶、磷酸核糖基-AMP环化水解酶、5’ProFAR异构酶、咪唑-甘油磷酸合酶、咪唑-甘油磷酸脱水酶、组氨醇-磷酸氨基转移酶、组氨醇-磷酸磷酸酶、组氨醇脱氢酶和磷酸核糖焦磷酸激酶，包括具有这些酶活性的任何同工型(isoform)、旁系同源物或直系同源物(本领域技术人员易于理解，它们可称为不同的名称)。所述至少一种其他改变可通过任何可获得的方式提高上游途径酶的活性，例如通过：(1)调节天然酶的表达或活性，(2)表达天然酶的基因的一个或多个额外拷贝，和/或(3)表达一种或多种非天然酶的基因的一个或多个拷贝。
在一些实施方案中，可通过表达一种或多种编码反馈失调的酶的基因来实现经由所述途径的通量的增加，如上文所讨论。例如，经改造的宿主细胞可包含并表达一种或多种反馈失调的ATP磷酸核糖基转移酶基因。
经改造的微生物细胞可包含引入的基因，其具有天然的核苷酸序列或不同于天然的核苷酸序列。例如，可对天然的核苷酸序列进行密码子优化，用于在特定宿主细胞中表达。由任何这些引入的基因编码的氨基酸序列可以是天然的或可以不同于天然的序列。在各实施方案中，氨基酸序列与天然的氨基酸序列具有至少60％、70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。
在一些实施方案中，通过降低消耗一种或多种组胺途径前体的酶(例如烯醇化酶、丙酮酸脱氢酶、磷酸戊糖途径糖异构酶、转醛醇酶、转酮醇酶、核酮糖-5-磷酸差向异构酶和核酮糖-5-磷酸异构酶(aribulose-5-phosphate isomerase))的表达或活性，可以实现至组胺的前体的提高的可用性。例如，经改造的宿主细胞可以包括一个或多个启动子交换(swap)以下调任何这些酶的表达和/或可以缺失它们的基因以完全消除它们的表达。
本文所述方法在细菌细胞即谷氨酸棒状杆菌和枯草芽孢杆菌(原核细胞)和真菌细胞即酿酒酵母和解脂亚罗酵母(真核细胞)中进行。(参见实施例1和2。)
示例性经改造的酵母细胞
在某些实施方案中，经改造的酵母(例如酿酒酵母)细胞表达异源的组胺脱羧酶，其与来自色杆菌属种LK1的组胺脱羧酶(如SEQ ID NO：6)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，色杆菌属种LK1组胺脱羧酶可包含SEQ ID NO：6。经改造的酵母(例如酿酒酵母)细胞还可表达异源的ATP磷酸核糖基转移酶，其与来自酿酒酵母的ATP磷酸核糖基转移酶(SEQ ID NO：3)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，酿酒酵母ATP磷酸核糖基转移酶包含SEQ ID NO：3。
在某些实施方案中，经改造的酵母(例如解脂亚罗酵母)细胞表达异源的组胺脱羧酶，其与来自鲍氏不动杆菌菌株AB0057的组胺脱羧酶(如SEQ ID NO：1)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，鲍氏不动杆菌菌株AB0057组胺脱羧酶可包含SEQ ID NO：1。经改造的酵母(例如解脂亚罗酵母)细胞还可表达异源的ATP磷酸核糖基转移酶，其与来自酿酒酵母S288c的ATP磷酸核糖基转移酶(SEQID NO：3)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，酿酒酵母S288c ATP磷酸核糖基转移酶包含SEQ ID NO：3。
这些可以是经改造的酵母细胞的唯一遗传改变，或者所述酵母细胞可包含一个或多个额外的遗传改变，如上文更一般性的讨论。
例如，在具体实施方案中，上述经改造的酵母酿酒酵母细胞额外地表达谷氨酸棒状杆菌ATP磷酸核糖基转移酶的反馈失调的变体，其通常与含有氨基酸置换N215K、L231F和T235A的谷氨酸棒状杆菌ATP磷酸核糖基转移酶的变体(SEQ ID NO：7)具有至少70％、75％、80％、85％、90％或95％氨基酸序列同一性。在具体实施方案中，谷氨酸棒状杆菌ATP磷酸核糖基转移酶变体可包含SEQ ID NO：7。
示例性经改造的细菌细胞
在某些实施方案中，经改造的细菌(例如谷氨酸棒状杆菌)细胞表达异源的组胺脱羧酶，其与来自鲍氏不动杆菌的组胺脱羧酶(如SEQ ID NO：1)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，鲍氏不动杆菌组胺脱羧酶可包含SEQ ID NO：1。经改造的细菌(例如谷氨酸棒状杆菌)细胞还可表达异源的ATP磷酸核糖基转移酶，其与来自酿酒酵母S288c的ATP磷酸核糖基转移酶(SEQ ID NO：3)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，酿酒酵母S288c ATP磷酸核糖基转移酶包含SEQ ID NO：3。在一些实施方案中，经改造的细菌(例如谷氨酸棒状杆菌)细胞表达咪唑-甘油磷酸脱水酶，而不是ATP磷酸核糖基转移酶，所述咪唑-甘油磷酸脱水酶与来自谷氨酸棒状杆菌的咪唑-甘油磷酸脱水酶(SEQ ID NO：2)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，谷氨酸棒状杆菌咪唑-甘油磷酸脱水酶包含SEQ ID NO：2。
在某些实施方案中，经改造的细菌(例如枯草芽孢杆菌)细胞表达异源的组胺脱羧酶，其与来自乳杆菌属种(菌株30a)的组胺脱羧酶(如SEQ ID NO：4)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，乳杆菌属种(菌株30a)组胺脱羧酶可包含SEQ ID NO：4。经改造的细菌(例如枯草芽孢杆菌)细胞还可表达异源的ATP磷酸核糖基转移酶，其与来自鼠伤寒沙门氏菌LT2的ATP磷酸核糖基转移酶(SEQ IDNO：5)具有至少70％、75％、80％、85％、90％、95％或100％氨基酸序列同一性。在具体实施方案中，鼠伤寒沙门氏菌LT2 ATP磷酸核糖基转移酶包含SEQ ID NO：5。
培养经改造的微生物细胞
可以培养本文所述的任何微生物细胞，例如用于维持、生长和/或产生组胺。
在一些实施方案中，培养物生长到600nm下的光密度为10-500，例如光密度为50-150。
在各实施方案中，培养物包括产生的组胺，其效价为至少10、25、50、75、100、200、300、400、500、600、700、800或900μg/L，或至少1、10、50、75、100、200、300、400、500、600、700、800或900mg/L或至少1、1.5、2、2.5、3、3.5、4、4.5、5、10、20、50g/L。在各实施方案中，效价范围为10μg/L至10g/L、25μg/L至20g/L、100μg/L至10g/L、200μg/L至5g/L、500μg/L至4g/L、1mg/L至3g/L、500mg/L至2g/L，或以上列出的任何数值界定的任何范围内。
培养基
微生物细胞可以在任何适合的培养基中培养，所述培养基包括但不限于基本培养基，即包含可用于细胞生长的最低营养成分的培养基。基本培养基通常包含：(1)用于微生物生长的碳源；(2)盐，其可取决于具体的微生物细胞和生长条件；和(3)水。适合的培养基还包含以下成分的任意组合：用于生长和产物形成的氮源，用于生长的硫源，用于生长的磷酸盐源，用于生长的金属盐，用于生长的维生素，以及用于生长的其他辅因子。
任何适合的碳源均可用于培养宿主细胞。术语“碳源”指一种或多种能够被微生物细胞代谢的含碳化合物。在各实施方案中，碳源是碳水化合物(例如单糖、二糖、寡糖或多糖)或转化糖(例如酶处理的蔗糖浆)。示例性单糖包括葡萄糖(右旋糖)、果糖(左旋糖)和半乳糖；示例性寡糖包括葡聚糖(dextran或glucan)；以及示例性多糖包括淀粉和纤维素。适合的糖包括C6糖(例如果糖、甘露糖、半乳糖或葡萄糖)和C5糖(例如木糖或阿拉伯糖)。其他更便宜的碳源包括甘蔗汁、甜菜汁、高粱汁等，任何这些都可以(但不需要)完全或部分去离子化。
培养基中的盐通常提供必需元素，例如镁、氮、磷和硫，以使细胞合成蛋白质和核酸。
基本培养基可补充一种或多种选择性试剂，例如抗生素。
为了产生组胺，培养基可包含和/或在培养中补充葡萄糖和/或氮源，例如尿素、铵盐、氨或其任意组合。
培养条件
适于微生物细胞的维持和生长的材料和方法是本领域公知的。参见，例如美国公开号2009/0203102、2010/0003716和2010/0048964，以及国际公开号WO 2004/033646、WO2009/076676、WO 2009/132220和WO 2010/003007，Manual of Methods for GeneralBacteriology，Gerhardt等人编著，American Society for Microbiology，Washington，D.C.(1994)或Brock in Biotechnology：A Textbook of Industrial Microbiology，第二版(1989)Sinauer Associates，Inc.，Sunderland，Mass。
通常，细胞生长并维持在适当的温度、气体混合物和pH下(例如约20℃至约37℃，约6％至约84％CO2，pH为约5至约9)。在一些方面中，细胞生长于35℃下。在某些实施方案中，例如当嗜热细菌用作宿主细胞时，可以使用更高的温度(例如50℃-75℃)。在一些方面中，用于发酵的pH范围为约pH 5.0至约pH 9.0(例如约pH 6.0至约pH 8.0或约6.5至约7.0)。根据具体细胞的需求，细胞可生长在有氧、缺氧或厌氧条件下。
可使用的标准培养条件和发酵模式，例如分批发酵、分批补料发酵或连续发酵记载于美国公开号2009/0203102、2010/0003716和2010/0048964，以及国际公开号WO 2009/076676、WO 2009/132220和WO 2010/003007中。分批发酵和分批补料发酵是本领域中常见和公知的，实例可参见Brock，Biotechnology：A Textbook of Industrial Microbiology，第二版(1989)Sinauer Associates，Inc.。
在一些实施方案中，将细胞培养在有限糖(例如葡萄糖)的条件下。在各实施方案中，添加的糖的量少于或约为细胞可消耗的糖的量的105％(例如约100％、90％、80％、70％、60％、50％、40％、30％、20％或10％)。在具体实施方案中，添加到培养基的糖的量与特定时间段内被细胞消耗的糖的量大约相同。在一些实施方案中，通过限制添加的糖的量来控制细胞生长速率，从而使细胞以细胞培养基中糖的量能够支持的速率生长。在一些实施方案中，糖在培养细胞期间不积累。在各实施方案中，在有限糖条件下培养细胞，培养时间长于或约为1、2、3、5、10、15、20、25、30、35、40、50、60或70小时，或甚至最长达约5-10天。在各实施方案中，在有限糖的条件下培养细胞，培养时间长于或约为培养细胞的总时长的5、10、15、20、25、30、35、40、50、60、70、80、90、95或100％。尽管不意欲受限于任何具体理论，认为有限糖的条件可以更有利地调节细胞。
在一些方面中，细胞生长于分批培养物中。细胞还可以生长于分批补料培养物或连续培养物中。另外，细胞可培养于基本培养基中，包括但不限于上文所述的任何基本培养基。基本培养基可进一步补充1.0％(w/v)或更少的葡萄糖(或任何其他六碳糖)。特别地，基本培养基可补充1％(w/v)、0.9％(w/v)、0.8％(w/v)、0.7％(w/v)、0.6％(w/v)、0.5％(w/v)、0.4％(w/v)、0.3％(w/v)、0.2％(w/v)或0.1％(w/v)葡萄糖。在一些培养中，使用显著更高水平的糖(例如葡萄糖)，例如至少10％(w/v)、20％(w/v)、30％(w/v)、40％(w/v)、50％(w/v)、60％(w/v)、70％(w/v)，或最高达所述糖在培养基中的溶解度极限。在一些实施方案中，糖水平落入以上数值中任意两个的范围内，例如0.1-10％(w/v)、1.0-20％(w/v)、10-70％(w/v)、20-60％(w/v)或30-50％(w/v)。此外，不同的糖水平可用于不同的培养阶段。对于(例如酿酒酵母或谷氨酸棒状杆菌的)分批补料培养，分批阶段的糖水平可以为约100-200g/L(10-20％(w/v))，之后最高达约500-700g/L(在补料中50-70％)。
另外，基本培养基可补充0.1％(w/v)或更少的酵母提取物。特别地，基本培养基可补充0.1％(w/v)、0.09％(w/v)、0.08％(w/v)、0.07％(w/v)、0.06％(w/v)、0.05％(w/v)、0.04％(w/v)、0.03％(w/v)、0.02％(w/v)或0.01％(w/v)酵母提取物。或者，基本培养基可补充1％(w/v)、0.9％(w/v)、0.8％(w/v)、0.7％(w/v)、0.6％(w/v)、0.5％(w/v)、0.4％(w/v)、0.3％(w/v)、0.2％(w/v)或0.1％(w/v)葡萄糖并补充0.1％(w/v)、0.09％(w/v)、0.08％(w/v)、0.07％(w/v)、0.06％(w/v)、0.05％(w/v)、0.04％(w/v)、0.03％(w/v)或0.02％(w/v)酵母提取物。在一些培养中，可使用显著更高水平的酵母提取物，例如至少1.5％(w/v)、2.0％(w/v)、2.5％(w/v)或3％(w/v)。在(例如酿酒酵母或谷氨酸棒状杆菌的)一些培养中，酵母提取物水平落入以上数值中任意两个的范围内，例如0.5-3.0％(w/v)、1.0-2.5％(w/v)或1.5-2.0％(w/v)。
适用于本文所述的经改造的微生物细胞的维持和生长的示例性材料和方法可参见以下实施例1。
组胺产生和回收
本文所述的任何方法还包括回收组胺的步骤。在一些实施方案中，从生产容器回收/收获包含于所谓的收获流(harvest stream)中的所产生的组胺。收获流可包括，例如从生产容器流出的无细胞或含细胞水溶液，其包含由生产容器中的静息细胞(resting cell)转化生产底物而得到的组胺。可通过本领域中已知的任何操作将仍存在于收获流中的细胞与组胺分离，所述操作例如过滤、离心、倾析(decantation)、膜错流超滤或微滤、切向流超滤或微滤或死端过滤。在该细胞分离操作之后，收获流基本上不含细胞。
可任选地进行产生的组胺从收获流中包含的其他组分中分离和/或纯化的其他步骤，即所谓的下游处理步骤。这些步骤可包括技术人员已知的任何方式，例如浓缩、萃取、结晶、沉淀、吸附、离子交换和/或色谱法。可单独使用这些步骤中的任何步骤，或将其组合使用以纯化组胺。其他纯化步骤可包括以下方法中的一种或多种：例如，浓缩、结晶、沉淀、清洗和干燥、用活性炭处理、离子交换、纳滤和/或重结晶。适合的纯化方案的设计可取决于细胞、培养基、培养规模、生产容器等，并且在本领域技术水平范围内。
提供以下实施例的目的在于说明公开内容的多种实施方案，并且不意欲以任何方式限制本公开内容。本领域技术人员将能够识别其中的变化和其他用途，其涵盖于本公开内容的宗旨内，由权利要求的范围限定。
实施例1——经改造以产生组胺的谷氨酸棒状杆菌和酿酒酵母菌株的构建和选择
质粒/DNA设计
本研究中所有测试菌株用质粒DNA转化，所述质粒DNA是使用专用软件设计。质粒设计是特别针对本研究中改造的每一种宿主生物体的。通过标准DNA组装方法物理构建质粒DNA。然后，通过分别如下文所述的两种宿主特异性方法之一，将该质粒DNA用于整合代谢途径插入。
谷氨酸棒状杆菌途径整合
“环入，单交换”基因组整合策略被开发以改造谷氨酸棒状杆菌菌株。图14举例说明仅环入的构建体和环入/环出构建体的基因组整合以及通过菌落PCR确认正确的整合。仅环入的构建体(在标题“环入”下示出)包含单个2kb同源臂(表示为“整合基因座”)、正向选择标记物(表示为“标记物”)和目的基因(表示为“启动子-基因-终止子”)。单交换事件将质粒整合到谷氨酸棒状杆菌染色体中。通过在存在抗生素(例如25μg/ml卡那霉素)的情况下生长，整合事件在基因组中稳定维持。通过使用UF/IR和DR/IF PCR引物的菌落PCR，确定源自环入整合的菌落中正确的基因组整合。
环入、环出(loop-in，loop-out)构建体(在标题“环入、环出”下示出)包含两个2kb同源臂(5’和3’臂)、目的基因(箭头)、正向选择标记物(表示为“标记物”)和反向选择标记物。类似于仅“环入”的构建体，单交换事件将质粒整合到谷氨酸棒状杆菌染色体中。注意：此处仅示出两个可能的整合之一。通过菌落PCR来确定正确的基因组整合，并且利用反向选择，使得可以切开质粒骨架和反向选择标记物。这导致两种可能性之一：回复至野生型(下左框)或所需的途径整合(下右框)。同样，通过菌落PCR来确定正确的基因组环出。(缩写：引物：UF＝上游正向，DR＝下游反向，IR＝内部反向，IF＝内部正向。)
酿酒酵母途径整合
“分离标记物，双交换(split-marker，double-crossover)”基因组整合策略被开发以改造酿酒酵母菌株。图11举例说明在酿酒酵母中，互补的分离标记物质粒的基因组整合以及通过菌落PCR确认正确的基因组整合。将具有互补的5’和3’同源臂和URA3选择性标记物的重叠部分(以斜线柱(hashed bar)表示直接重复)的两种质粒用大范围核酸酶(meganuclease)消化并作为线性片段转化。三重交换事件将所需的异源基因整合到靶基因座并重新构建完整的URA3基因。使用两个3-引物反应来测定源自该整合事件的菌落以确定5’和3’接合(UF/IF/wt-R和DR/IF/wt-F)。对于需要进一步改造的菌株，可将菌株铺在5-FOA平板上以选择URA3去除，留下小的单拷贝的原始直接重复。该基因组整合策略可用于相同操作流程中的基因敲除、基因敲入和启动子滴定(promoter titration)。
细胞培养
为酿酒酵母建立的操作流程包括命中挑选(hit-picking)步骤，该步骤使用在平板上随机化菌株的自动化操作流程来强化成功构建的菌株。对于每种成功构建的菌株，从不同的菌落测试最多达4个重复，以测试菌落之间差异和其他过程差异。如果获得的菌落少于4个，则重复现有菌落，使得从每个需要的基因型测试至少4个孔。
将菌落合并到具有选择性培养基(用于酿酒酵母的SD-ura)的96孔板并培养两天直至饱和，然后用16.6％甘油在-80℃下冷冻保存。然后将冷冻的甘油原液用于在基本培养基中接种种子期，所述基本培养基具有低水平氨基酸以有利于生长和从冷冻中恢复。种子平板在30℃下生长1-2天。然后将种子平板用于接种具有基本培养基的主要培养板并生长48-88小时。在需要的时间点移出平板并测试细胞密度(OD600)、活力和葡萄糖，贮存的上清液样品用于目的产物的LC-MS分析。
细胞密度
使用分光光度测定法来测量细胞密度，所述测定检测600nm下各孔的吸光度。机器人技术用于将确定量的培养物从各培养板转移到测定板，然后与175mM磷酸钠(pH 7.0)混合以产生10倍稀释液。使用Tecan M1000分光光度仪测量测定板，并将测定数据上传至LIMS数据库。未接种对照用于减去背景吸光度。通过以下方式来监测细胞生长：在每个时期接种多个平板，然后在每个时间点放弃(sacrifice)整个平板。
为了最小化细胞沉淀同时处理大量平板(这在测量期间可导致非代表性样品)，在每次读数之前振荡每个平板10-15秒。平板内细胞密度的大幅度差异还可导致检测线性范围以外的吸光度测量值，导致对较高OD培养物的低估。总体上，迄今为止测试的菌株的差异尚未显著到足以成为问题的程度。
液体-固体分离
为了收集胞外样品用于LC-MS分析，通过离心分离固相和液相。2000rpm离心培养板4分钟，使用机器人技术将上清液转移到目标板。将75μL上清液转移到各板，其中一个贮存于4℃，第二个贮存于80℃以长期保存。
谷氨酸棒状杆菌和酿酒酵母中的第一轮基因工程结果
采用文库方法筛选异源途径酶以建立组胺途径。对于组氨酸脱羧酶，从表1中列出的细菌、古细菌、绿色植物(Viridiplantae)、脊椎动物门、后生动物和节肢动物门来源中测试了18个异源序列。对组氨酸脱羧酶进行密码子优化，并在酿酒酵母和谷氨酸棒状杆菌宿主中表达。
组氨酸生物合成受到反馈抑制，因此用组氨酸脱羧酶测试了反馈失调的ATP磷酸核糖基转移酶，以提高组氨酸脱羧酶底物组氨酸的产生。测试的ATP磷酸核糖基转移酶来自鼠伤寒沙门氏菌和谷氨酸棒状杆菌，携带已知的缺失和点突变，使它们对反馈抑制产生抗性。
第一轮基因工程结果示于表1和图2(谷氨酸棒状杆菌)和图3(酿酒酵母)。
Figure BDA0002807770440000321
Figure BDA0002807770440000331
Figure BDA0002807770440000341
Figure BDA0002807770440000351
Figure BDA0002807770440000361
谷氨酸棒状杆菌和酿酒酵母的第二轮基因工程结果
采用文库方法，通过用组成型启动子分别表达每个上游途径酶来筛选限速步骤从而提高组胺的产生。筛选的组氨酸途径酶列于表2。此外，对于表2中的酶，菌株含有来自第一轮的最佳酶：谷氨酸棒状杆菌宿主含有组氨酸脱羧酶(UniProt ID B71459)(SEQ ID NO：1)和ATP磷酸核糖基转移酶(UniProt ID P00499)(SEQ ID NO：5)，该ATP磷酸核糖基转移酶含有缺失Q207-E208；以及酿酒酵母宿主含有组氨酸脱羧酶(UniProt ID J6KM89)(SEQ IDNO：6)和ATP磷酸核糖基转移酶(UniProt ID Q9Z472)(SEQ ID NO：7)，该ATP磷酸核糖基转移酶含有氨基酸置换N215K、L231F和T235A。
第二轮基因工程结果示于表2和图4(谷氨酸棒状杆菌)和图5(酿酒酵母)。
在谷氨酸棒状杆菌中，通过整合两个基因(来自鲍氏不动杆菌的组氨酸脱羧酶基因和来自谷氨酸棒状杆菌的咪唑甘油-磷酸脱水酶的组成型表达)的两轮改造后，获得了24mg/L的效价。
在酿酒酵母中，通过整合三个基因(来自色杆菌属种LK1的组氨酸脱羧酶基因(SEQID NO：6)、含有氨基酸置换N215K、L231F和T235A的来自谷氨酸棒状杆菌的ATP磷酸核糖基转移酶(SEQ ID NO：7)和来自酿酒酵母的组成型表达的ATP磷酸核糖基转移酶(SEQ ID NO：3))在两轮改造中，获得了385mg/L的效价。
Figure BDA0002807770440000381
Figure BDA0002807770440000391
Figure BDA0002807770440000401
酿酒酵母中的第三轮基因工程结果
在酿酒酵母中进一步追求组胺的产生，并且我们设计了质粒来整合由强的组成型启动子表达的上游途径基因的额外拷贝，以避免基因的天然调控(表3)。进行了扩展搜索以测试其他组氨酸脱羧酶，其序列与最初确定为活性的酶的序列相似(表3)。
同时，我们追求调节天然基因的表达以进一步提高组胺的产生。我们的改造方法是从第二轮中获得最佳的酿酒酵母菌株，并测试强的或弱的组成型启动子以代替天然启动子。选择启动子改变的基因靶标以将通量供应前体改向到组氨酸。被测试的菌株设计包括通过表达非天然的反馈失调的葡萄糖-6-磷酸脱氢酶(zwf)来增加戊糖磷酸途径通量和通过糖异构酶来减少“较低”的戊糖磷酸途径通量的设计。
对于被认为是必要的(即不能缺失的)，预计将增加可用于组胺产生的上部糖酵解代谢物池的低表达基因，启动子替换针对：1)烯醇化酶(Eno2)，通过较低的糖酵解减少通量；2)丙酮酸脱氢酶(PDH，Lpd1)，通过C3/C2节点减少通量；和3)戊糖磷酸途径糖异构酶，其使用组胺代谢物前体核糖-5-磷酸(Tal1)。酿酒酵母中的启动子交换(“proswap”)和缺失(“敲除”)靶标的示例性列表包括：
Figure BDA0002807770440000411
Figure BDA0002807770440000421
根据Lee等人[7]的表达数据选择启动子。
酿酒酵母的其他基因工程结果示于表3和图10。表3所示的菌株设计的亲本菌株(也是参考菌株，ScHISMN_41)含有组氨酸脱羧酶(UniProt ID J6KM89)和携带氨基酸置换N215K、L231F和T235A的ATP磷酸核糖基转移酶(UniProt ID Q9Z472)，以及来自酿酒酵母的ATP磷酸核糖基转移酶。参考菌株的组胺效价为131mg/L。
在从强的组成型启动子表达下列每种酶的菌株中测定出提高的效价：
1.转酮醇酶(EC 2.2.1.1)(SEQ ID NO：27)，其催化戊糖磷酸途径中糖的相互转化，并产生核糖-5-磷酸，这是组氨酸生物合成途径中最初的代谢物PPRP的前体。
2.核糖-磷酸焦磷酸激酶(EC 2.7.6.1)(SEQ ID NO：28)(最高效价：191mg/L，相对于实验中的对照131mg/L)。
3.ATP磷酸核糖基转移酶(EC 2.4.2.17)(SEQ ID NO：3)。
4.三功能的组氨醇脱氢酶(EC 1.1.1.23)/磷酸核糖基-AMP环化水解酶(EC3.5.4.19)/磷酸核糖基-ATP二磷酸酶(EC 3.6.1.31)(SEQ ID NO：29)。
5.组氨醇-磷酸氨基转移酶(EC 2.6.1.9)(SEQ ID NO：14)。
6.5’ProFAR异构酶(EC 5.3.1.16)(SEQ ID NO：31)。
7.咪唑甘油磷酸合酶(EC 4.3.1.B2)(SEQ ID NO：21)。
8.携带氨基酸置换I170V(SEQ ID NO：32)或I170T[8]的丙糖-磷酸异构酶(EC5.3.1.1)。
9.携带氨基酸置换A243T的葡萄糖-6-磷酸1-脱氢酶(EC 1.1.1.49)(SEQ ID NO：26)。
10.各种组氨酸脱羧酶(EC 4.1.1.22)：
a.UniProt ID A0A089YPE5(SEQ ID NO：33)
b.UniProt ID A0A126S6G9(SEQ ID NO：34)
c.UniProt ID A0A0A1R6V3(SEQ ID NO：35)
d.UniProt ID A0A1W0CM88(SEQ ID NO：36)
e.UniProt ID P00862(SEQ ID NO：4)
f.UniProt ID A0A0K6GJ74(SEQ ID NO：37)
g.UniProt ID T0QL99(SEQ ID NO：38)
h.UniProt ID A0A1B8HLR1(SEQ ID NO：39)
Figure BDA0002807770440000441
Figure BDA0002807770440000451
Figure BDA0002807770440000461
Figure BDA0002807770440000471
Figure BDA0002807770440000481
Figure BDA0002807770440000491
Figure BDA0002807770440000501
Figure BDA0002807770440000511
Figure BDA0002807770440000521
Figure BDA0002807770440000531
Figure BDA0002807770440000541
实施例2-组胺产生的宿主评估
还在两个另外的宿主(枯草芽孢杆菌和解脂亚罗酵母)中测试组胺产生，所述宿主经改造以表达来自最佳效能谷氨酸棒状杆菌和酿酒酵母菌株的酶。
选择宿主评估设计来表达1-3种酶，并根据宿主生物体谷氨酸棒状杆菌、酿酒酵母、枯草芽孢杆菌和解脂亚罗酵母，对每种设计进行四种不同密码子优化测试。测试的密码子优化是基于针对每个宿主制表的Kazusa密码子使用表，以进行基因密码子优化(www.kazusa.or.jp/codon/)。
在解脂亚罗酵母(图6)和枯草芽孢杆菌(图7)中证明了组胺的产生，以及在谷氨酸棒状杆菌(图9)和酿酒酵母(图8)中进一步提高了组胺的产生。
在解脂亚罗酵母中(以下图6，表4)，最佳效能的菌株产生505mg/L组胺，并表达来自鲍氏不动杆菌菌株AB0057的组氨酸脱羧酶(UniProt ID B71459)和来自酿酒酵母S288c的ATP磷酸核糖基转移酶(UniProt ID P00498)，其中组氨酸脱羧酶和ATP磷酸核糖基转移酶的DNA序列针对解脂亚罗酵母进行密码子优化。还测试了同样的两个基因，其中DNA序列针对枯草芽孢杆菌和酿酒酵母进行密码子优化，所得菌株没有产生组胺效价。
在解脂亚罗酵母中，第二佳效能的菌株还表达来自鲍氏不动杆菌菌株AB0057的组氨酸脱羧酶(UniProt ID B71459)和来自鼠伤寒沙门氏菌LT2的ATP磷酸核糖基转移酶(UniProt ID P00499)，其中组氨酸脱羧酶和ATP磷酸核糖基转移酶的DNA序列针对解脂亚罗酵母进行密码子优化。还测试了这两个基因的版本，其中DNA序列针对枯草芽孢杆菌进行密码子优化(其产生0效价)，DNA序列针对酿酒酵母进行密码子优化(其产生33微克组胺)，以及使用酿酒酵母和谷氨酸棒状杆菌的组合的密码子表对DNA序列进行密码子优化(产生97mg/L组胺)。
在解脂亚罗酵母中，第三佳效能的菌株产生258mg/L组胺，并表达来自色杆菌属种LK1的组氨酸脱羧酶(UniProt ID A0A0J6KM89)和来自谷氨酸棒状杆菌ATCC 13032的携带氨基酸置换N215K、L231F、T235A的ATP磷酸核糖基转移酶(UniProt ID Q9Z472)(SEQ IDNO：7)，其中组氨酸脱羧酶的DNA序列针对解脂亚罗酵母进行密码子优化，ATP磷酸核糖基转移酶的DNA序列针对解脂亚罗酵母进行密码子优化(SEQ ID NO：64)。还测试了这两个基因的版本，其中DNA序列针对酿酒酵母(SEQ ID NO：65、66)或枯草芽孢杆菌(SEQ ID NO：67、68)进行密码子优化，这些解脂亚罗酵母菌株分别产生1.8mg/L和0.3mg/L。因此，基因的密码子优化影响解脂亚罗酵母中的表达。
在枯草芽孢杆菌中(以下图7，表5)，最佳效能的菌株产生18mg/L组胺，并表达来自乳杆菌属种(菌株30a)的组胺脱羧酶(UniProt ID P00862)(SEQ ID NO：4)和来自鼠伤寒沙门氏菌LT2的ATP磷酸核糖基转移酶(UniProt ID P00499)(SEQ ID NO：5)，其中DNA序列针对枯草芽孢杆菌进行密码子优化(SEQ ID NO：69、59)。还测试了同样的两个基因，其中DNA序列针对酿酒酵母进行密码子优化(SEQ ID NO：70、60)或使用谷氨酸棒状杆菌和酿酒酵母的改良的密码子使用表对DNA序列进行密码子优化(SEQ ID NO：71、62)，这些菌株分别产生6.7mg/L或0mg/L组胺。
还在酿酒酵母和谷氨酸棒状杆菌中测试了宿主评估设计。在酿酒酵母中(以下图8，表6)，最佳效能的菌株产生111mg/L组胺，并表达来自色杆菌属种LK1的组胺脱羧酶(UniProt ID A0A0J6KM89)(SEQ ID NO：51)和来自酿酒酵母S288c的ATP磷酸核糖基转移酶(UniProt ID P00498)(SEQ ID NO：3)，其中DNA序列针对解脂亚罗酵母进行密码子优化(SEQ ID NO：63、53)。还测试了同样的两个基因，其中DNA序列针对酿酒酵母(SEQ ID NO：65、57)和枯草芽孢杆菌(SEQ ID NO：67、55)进行密码子优化，这些菌株分别产生86mg/L和101mg/L。
在谷氨酸棒状杆菌中(图9，表7)，最佳效能的菌株产生68mg/L组胺，并表达来自鲍氏不动杆菌(菌株AB0057)的组胺脱羧酶(UniProt ID B71459)(SEQ ID NO：1)和来自酿酒酵母S288c的ATP磷酸核糖基转移酶(UniProt ID P00498)(SEQ ID NO：3)，其中使用谷氨酸棒状杆菌和酿酒酵母的改良的密码子使用表对DNA序列进行密码子优化(SEQ ID NO：72、73)。还测试了同样的两个基因，其中DNA序列针对解脂亚罗酵母(SEQ ID NO：52、53)或酿酒酵母(SEQ ID NO：56、57)进行密码子优化，这些菌株分别产生16mg/L和18微克/L组胺。
在谷氨酸棒状杆菌中，第二佳效能的菌株产生15mg/L组胺，还表达来自鲍氏不动杆菌菌株AB0057的组氨酸脱羧酶(UniProt ID B7I459)(SEQ ID NO：1)和来自鼠伤寒沙门氏菌LT2的ATP磷酸核糖基转移酶(UniProt ID P00499)(SEQ ID NO：5)，其中组氨酸脱羧酶的DNA序列针对解脂亚罗酵母进行密码子优化(SEQ ID NO：52)，ATP磷酸核糖基转移酶的DNA针对解脂亚罗酵母进行密码子优化(SEQ ID NO：58)。还测试了同样的这两个基因，其中DNA序列针对枯草芽孢杆菌进行密码子优化(SEQ ID NO：54、59)(其产生8mg/L组胺)或针对酿酒酵母进行密码子优化(SEQ ID NO：56、60)(其产生9.3mg/L组胺)。
由于最佳效能的菌株是在宿主解脂亚罗酵母中，可以在该宿主生物体中追求进一步的菌株改良。可以进一步增强解脂亚罗酵母中组胺产生的设计包括：
1.转酮醇酶(EC 2.2.1.1)(SEQ ID NO：27)，其催化戊糖磷酸途径中糖的相互转化，并产生核糖-5-磷酸，这是组氨酸生物合成途径中最初的代谢物PPRP的前体。
2.核糖-磷酸焦磷酸激酶(EC 2.7.6.1)(SEQ ID NO：28)。
3.ATP磷酸核糖基转移酶(EC 2.4.2.17)(SEQ ID NO：5)。
4.三功能的组氨醇脱氢酶(EC 1.1.1.23)/磷酸核糖基-AMP环化水解酶(EC3.5.4.19)/磷酸核糖基-ATP二磷酸酶(EC 3.6.1.31)(SEQ ID NO：20)。
5.组氨醇-磷酸氨基转移酶(EC 2.6.1.9)(SEQ ID NO：14)。
6.5’ProFAR异构酶(EC 5.3.1.16)(SEQ ID NO：31)。
7.咪唑甘油磷酸合酶(EC 4.3.1.B2)(SEQ ID NO：21)。
8.携带氨基酸置换I170V的丙糖-磷酸异构酶(EC 5.3.1.1)(SEQ ID NO：32)。
9.携带氨基酸置换A243T的葡萄糖-6-磷酸1-脱氢酶(EC 1.1.1.49)(SEQ ID NO：26)。
10.各种组氨酸脱羧酶：
a.UniProt ID A0A089YPE5(SEQ ID NO：33)
b.UniProt ID A0A126S6G9(SEQ ID NO：34)
c.UniProt ID A0A0A1R6V3(SEQ ID NO：35)
d.UniProt ID A0A1W0CM88(SEQ ID NO：36)
e.UniProt ID P00862(SEQ ID NO：4)
f.UniProt ID A0A0K6GJ74(SEQ ID NO：37)
g.UniProt ID T0QL99(SEQ ID NO：38)
h.UniProt ID A0A1B8HLR1(SEQ ID NO：39)
实施例3-经改造以产生组胺的解脂亚罗酵母中组胺产生的提高
在解脂亚罗酵母中进行了三轮提高轮基因工程。
在解脂亚罗酵母中的第一提高轮基因工程
策略：通过两种酶的过表达，提高进入组氨酸和随后的组胺的通量。
Figure BDA0002807770440000581
综述：ATP磷酸核糖基转移酶催化组氨酸生物合成途径的第一个关键步骤(committed step)。这种酶将被组氨酸别构反馈抑制，并被AMP和ADP竞争性抑制。结果没有表明P00498的活性和/或抑制作用。
在解脂亚罗酵母中的第二提高轮基因工程
策略：过表达一种酶。通过利用第一轮最佳的组氨酸脱羧酶增强组胺生物合成的最后一步，该酶被修饰为包括溶解性标签以改善蛋白质折叠。
Figure BDA0002807770440000582
综述：第二轮基因工程使用的组氨酸脱羧酶与第一轮相同，尽管密码子优化不同。此外，第二轮酶中还包括N端溶解性标签(MQYKLALNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDDATKTFTVT，SEQ ID NO：142)。
在解脂亚罗酵母中的第三提高轮基因工程
策略：过表达组氨酸生物合成上游途径中的两种酶，以提高进入磷酸核糖焦磷酸(PRPP)的通量。
Figure BDA0002807770440000591
综述：核糖-磷酸焦磷酸激酶被ADP竞争性抑制。该酶上ATP结合位点的L135I突变缓解了ADP抑制作用。该菌株以1.68g/L培养基的效价表达组胺。
Figure BDA0002807770440000601
Figure BDA0002807770440000611
Figure BDA0002807770440000621
Figure BDA0002807770440000631
Figure BDA0002807770440000641
Figure BDA0002807770440000651
Figure BDA0002807770440000661
Figure BDA0002807770440000671
Figure BDA0002807770440000681
Figure BDA0002807770440000691
Figure BDA0002807770440000701
Figure BDA0002807770440000711
Figure BDA0002807770440000721
Figure BDA0002807770440000731
Figure BDA0002807770440000741
Figure BDA0002807770440000751
Figure BDA0002807770440000761
Figure BDA0002807770440000771
Figure BDA0002807770440000781
Figure BDA0002807770440000791
Figure BDA0002807770440000801
Figure BDA0002807770440000811
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6.Wauters，G.，et al.，Histidine decarboxylase in Enterobacteriaceaerevisited.J Clin Microbiol，2004.42(12)：p.5923-4.
7.Lee，M.E.，et al.，A Highly Characterized Yeast Toolkit for Modular，Multipart Assembly.ACS Synth Biol，2015.4(9)：p.975-86.
8.Roland，B.P.，et al.，Triosephosphate isomerase I170V alters catalyticsite，enhances stability and induces pathology in a Drosophila model of TPIdeficiency.Biochim Biophys Acta，2015.1852(1)：p.61-9.
序列表
<110> 齐默尔根公司
<120> 通过发酵产生组胺的经改造的生物合成途径
<130> ZMGNP011PWO
<150> US 62/660,875
<151> 2018-04-20
<160> 143
<170> PatentIn version 3.5
<210> 1
<211> 383
<212> PRT
<213> 鲍氏不动杆菌（Acinetobacter baumannii）（菌株AB0057）
<400> 1
Met Ile Leu Ser Pro Ala Asp Gln Glu Arg Ile Glu Thr Phe Trp Asn
1 5 10 15
Tyr Cys Leu Lys His Gln Tyr Phe Asn Ile Gly Tyr Pro Glu Ser Ala
20 25 30
Asp Phe Asp Tyr Ser Ala Leu Phe Arg Phe Phe Lys Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Lys Asp Tyr Ser Asn Tyr Ala Leu Asn Ser Phe
50 55 60
Asp Phe Glu Lys Asp Val Met Ala Tyr Phe Ala Glu Ile Phe Gln Ile
65 70 75 80
Pro Phe Glu Glu Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Ala Arg Glu Leu Phe Ser Asp Ser Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Thr His Tyr Ser Val Gly Lys Ile Ala Lys
115 120 125
Leu Leu Gln Met Lys Ser Cys Val Ile Glu Ser Leu Asp Asn Gly Glu
130 135 140
Ile Asp Tyr Asp Asp Leu Ile His Lys Ile Lys Thr Asn Lys Glu Ser
145 150 155 160
His Pro Ile Ile Phe Ala Asn Ile Gly Thr Thr Met Thr Gly Ala Ile
165 170 175
Asp Asp Ile Glu Met Ile Gln Glu Arg Leu Ala Gln Ile Gly Ile Met
180 185 190
Arg Arg Asp Tyr Tyr Ile His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asp His Pro Gln Ala Phe Ser Phe Ala His Gly Ile
210 215 220
Asp Ser Ile Cys Val Ser Gly His Lys Met Ile Gly Ser Pro Ile Pro
225 230 235 240
Cys Gly Ile Val Val Ala Lys Arg Gln Asn Val Glu Arg Ile Ser Val
245 250 255
Asp Val Asp Tyr Ile Ser Thr Arg Asp Gln Thr Ile Ser Gly Ser Arg
260 265 270
Asn Gly His Thr Val Leu Leu Met Trp Ala Ala Ile Arg Ser Gln Thr
275 280 285
Asn Leu Gln Arg Arg Gln Arg Ile Gln His Cys Leu Lys Met Ala Gln
290 295 300
Tyr Ala Val Asp Arg Phe Gln Ala Val Gly Ile Pro Ala Trp Arg Asn
305 310 315 320
Pro Asn Ser Ile Thr Val Val Phe Pro Cys Pro Ser Glu His Ile Trp
325 330 335
Lys Lys His Tyr Leu Ala Thr Ser Gly Asn Met Ala His Leu Ile Thr
340 345 350
Thr Ala His His Arg Asp Thr Arg Gln Ile Asp Ser Leu Ile Asp Asp
355 360 365
Val Ile Phe Asp Leu Gln Gly Ala Ser Lys Arg Thr Val Gly Phe
370 375 380
<210> 2
<211> 202
<212> PRT
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 2
Met Thr Val Ala Pro Arg Ile Gly Thr Ala Thr Arg Thr Thr Ser Glu
1 5 10 15
Ser Asp Ile Thr Val Glu Ile Asn Leu Asp Gly Thr Gly Lys Val Asp
20 25 30
Ile Asp Thr Gly Leu Pro Phe Phe Asp His Met Leu Thr Ala Phe Gly
35 40 45
Val His Gly Ser Phe Asp Leu Lys Val His Ala Lys Gly Asp Ile Glu
50 55 60
Ile Asp Ala His His Thr Val Glu Asp Thr Ala Ile Val Leu Gly Gln
65 70 75 80
Ala Leu Leu Asp Ala Ile Gly Asp Lys Lys Gly Ile Arg Arg Phe Ala
85 90 95
Ser Cys Gln Leu Pro Met Asp Glu Ala Leu Val Glu Ser Val Val Asp
100 105 110
Ile Ser Gly Arg Pro Tyr Phe Val Ile Ser Gly Glu Pro Asp His Met
115 120 125
Ile Thr Ser Val Ile Gly Gly His Tyr Ala Thr Val Ile Asn Glu His
130 135 140
Phe Phe Glu Thr Leu Ala Leu Asn Ser Arg Ile Thr Leu His Val Ile
145 150 155 160
Cys His Tyr Gly Arg Asp Pro His His Ile Thr Glu Ala Glu Tyr Lys
165 170 175
Ala Val Ala Arg Ala Leu Arg Gly Ala Val Glu Met Asp Pro Arg Gln
180 185 190
Thr Gly Ile Pro Ser Thr Lys Gly Ala Leu
195 200
<210> 3
<211> 297
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 3
Met Asp Leu Val Asn His Leu Thr Asp Arg Leu Leu Phe Ala Ile Pro
1 5 10 15
Lys Lys Gly Arg Leu Tyr Ser Lys Ser Val Ser Ile Leu Asn Gly Ala
20 25 30
Asp Ile Thr Phe His Arg Ser Gln Arg Leu Asp Ile Ala Leu Ser Thr
35 40 45
Ser Leu Pro Val Ala Leu Ile Phe Leu Pro Ala Ala Asp Ile Pro Thr
50 55 60
Phe Val Gly Glu Gly Lys Cys Asp Leu Gly Ile Thr Gly Val Asp Gln
65 70 75 80
Val Arg Glu Ser Asp Val Asp Val Asp Leu Ala Ile Asp Leu Gln Phe
85 90 95
Gly Asn Cys Lys Leu Gln Val Gln Val Pro Val Asn Gly Glu Tyr Lys
100 105 110
Lys Pro Glu Gln Leu Ile Gly Lys Thr Ile Val Thr Ser Phe Val Lys
115 120 125
Leu Ala Glu Lys Tyr Phe Ala Asp Leu Glu Gly Thr Thr Val Glu Lys
130 135 140
Met Thr Thr Arg Ile Lys Phe Val Ser Gly Ser Val Glu Ala Ser Cys
145 150 155 160
Ala Leu Gly Ile Gly Asp Ala Ile Val Asp Leu Val Glu Ser Gly Glu
165 170 175
Thr Met Arg Ala Ala Gly Leu Val Asp Ile Ala Thr Val Leu Ser Thr
180 185 190
Ser Ala Tyr Leu Ile Glu Ser Lys Asn Pro Lys Ser Asp Lys Ser Leu
195 200 205
Ile Ala Thr Ile Lys Ser Arg Ile Glu Gly Val Met Thr Ala Gln Arg
210 215 220
Phe Val Ser Cys Ile Tyr Asn Ala Pro Glu Asp Lys Leu Pro Glu Leu
225 230 235 240
Leu Lys Val Thr Pro Gly Arg Arg Ala Pro Thr Ile Ser Lys Ile Asp
245 250 255
Asp Glu Gly Trp Val Ala Val Ser Ser Met Ile Glu Arg Lys Thr Lys
260 265 270
Gly Val Val Leu Asp Glu Leu Lys Arg Leu Gly Ala Ser Asp Ile Met
275 280 285
Val Phe Glu Ile Ser Asn Cys Arg Val
290 295
<210> 4
<211> 311
<212> PRT
<213> 乳杆菌属种（Lactobacillus sp.）（菌株30a）
<400> 4
Met Ser Glu Leu Asp Ala Lys Leu Asn Lys Leu Gly Val Asp Arg Ile
1 5 10 15
Ala Ile Ser Pro Tyr Lys Gln Trp Thr Arg Gly Tyr Met Glu Pro Gly
20 25 30
Asn Ile Gly Asn Gly Tyr Val Thr Gly Leu Lys Val Asp Ala Gly Val
35 40 45
Arg Asp Lys Ser Asp Asp Asp Val Leu Asp Gly Ile Val Ser Tyr Asp
50 55 60
Arg Ala Glu Thr Lys Asn Ala Tyr Ile Gly Gln Ile Asn Met Thr Thr
65 70 75 80
Ala Ser Ser Phe Thr Gly Val Gln Gly Arg Val Ile Gly Tyr Asp Ile
85 90 95
Leu Arg Ser Pro Glu Val Asp Lys Ala Lys Pro Leu Phe Thr Glu Thr
100 105 110
Gln Trp Asp Gly Ser Glu Leu Pro Ile Tyr Asp Ala Lys Pro Leu Gln
115 120 125
Asp Ala Leu Val Glu Tyr Phe Gly Thr Glu Gln Asp Arg Arg His Tyr
130 135 140
Pro Ala Pro Gly Ser Phe Ile Val Cys Ala Asn Lys Gly Val Thr Ala
145 150 155 160
Glu Arg Pro Lys Asn Asp Ala Asp Met Lys Pro Gly Gln Gly Tyr Gly
165 170 175
Val Trp Ser Ala Ile Ala Ile Ser Phe Ala Lys Asp Pro Thr Lys Asp
180 185 190
Ser Ser Met Phe Val Glu Asp Ala Gly Val Trp Glu Thr Pro Asn Glu
195 200 205
Asp Glu Leu Leu Glu Tyr Leu Glu Gly Arg Arg Lys Ala Met Ala Lys
210 215 220
Ser Ile Ala Glu Cys Gly Gln Asp Ala His Ala Ser Phe Glu Ser Ser
225 230 235 240
Trp Ile Gly Phe Ala Tyr Thr Met Met Glu Pro Gly Gln Ile Gly Asn
245 250 255
Ala Ile Thr Val Ala Pro Tyr Val Ser Leu Pro Ile Asp Ser Ile Pro
260 265 270
Gly Gly Ser Ile Leu Thr Pro Asp Lys Asp Met Glu Ile Met Glu Asn
275 280 285
Leu Thr Met Pro Glu Trp Leu Glu Lys Met Gly Tyr Lys Ser Leu Ser
290 295 300
Ala Asn Asn Ala Leu Lys Tyr
305 310
<210> 5
<211> 297
<212> PRT
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 5
Met Leu Asp Asn Thr Arg Leu Arg Ile Ala Ile Gln Lys Ser Gly Arg
1 5 10 15
Leu Ser Asp Asp Ser Arg Glu Leu Leu Ala Arg Cys Gly Ile Lys Ile
20 25 30
Asn Leu His Thr Gln Arg Leu Ile Ala Met Ala Glu Asn Met Pro Ile
35 40 45
Asp Ile Leu Arg Val Arg Asp Asp Asp Ile Pro Gly Leu Val Met Asp
50 55 60
Gly Val Val Asp Leu Gly Ile Ile Gly Glu Asn Val Leu Glu Glu Glu
65 70 75 80
Leu Leu Asn Arg Arg Ala Gln Gly Glu Asp Pro Arg Tyr Leu Thr Leu
85 90 95
Arg Arg Leu Asp Phe Gly Gly Cys Arg Leu Ser Leu Ala Thr Pro Val
100 105 110
Asp Glu Ala Trp Asp Gly Pro Ala Ala Leu Asp Gly Lys Arg Ile Ala
115 120 125
Thr Ser Tyr Pro His Leu Leu Lys Arg Tyr Leu Asp Gln Lys Gly Val
130 135 140
Ser Phe Lys Ser Cys Leu Leu Asn Gly Ser Val Glu Val Ala Pro Arg
145 150 155 160
Ala Gly Leu Ala Asp Ala Ile Cys Asp Leu Val Ser Thr Gly Ala Thr
165 170 175
Leu Glu Ala Asn Gly Leu Arg Glu Val Glu Val Ile Tyr Arg Ser Lys
180 185 190
Ala Cys Leu Ile Gln Arg Asp Gly Glu Met Ala Gln Ser Lys Leu Ile
195 200 205
Asp Lys Leu Leu Thr Arg Ile Gln Gly Val Ile Gln Ala Arg Glu Ser
210 215 220
Lys Tyr Ile Met Met His Ala Pro Ser Glu Arg Leu Glu Glu Val Ile
225 230 235 240
Ala Leu Leu Pro Gly Ala Glu Arg Pro Thr Ile Leu Pro Leu Ala Gly
245 250 255
Glu Gln Gln Arg Val Ala Met His Met Val Ser Ser Glu Thr Leu Phe
260 265 270
Trp Glu Thr Met Glu Lys Leu Lys Ala Leu Gly Ala Ser Ser Ile Leu
275 280 285
Val Leu Pro Ile Glu Lys Met Met Glu
290 295
<210> 6
<211> 387
<212> PRT
<213> 色杆菌属种（Chromobacterium sp.）LK1
<400> 6
Met Ser Leu Ser Pro Leu Asp Gln Asn Arg Ile Glu Ser Phe Trp Gln
1 5 10 15
Tyr Cys Leu Gln His Gln Tyr Phe Asn Leu Ala Tyr Pro Glu Ser Ala
20 25 30
Asp Phe Asp Tyr Ala Pro Leu His Arg Phe Leu Arg Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Asn Glu Ser Ser Asn Tyr Leu Leu Asn Ser Phe
50 55 60
Asp Phe Glu Arg Glu Val Met His Phe Phe Ala Glu Leu Phe His Ile
65 70 75 80
Pro Phe Asp Glu Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Ala Arg Glu Leu Phe Pro Asp Ala Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Ser His Tyr Ser Val Ala Lys Ile Ile Lys
115 120 125
Leu Leu Arg Ile Lys Ser Arg Ala Val Asp Ser Leu Pro Ser Gly Glu
130 135 140
Ile Asp Tyr Asp Asp Leu Val Ala Lys Ile Gln Gln Asp Gln Glu Arg
145 150 155 160
His Pro Ile Val Phe Val Asn Val Gly Thr Thr Met Lys Gly Ala Val
165 170 175
Asp Asp Ile Gly Val Ile Gln His Lys Leu Ala Glu Ala Gly Ile Pro
180 185 190
Arg Gln Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asp Ala Pro Gln Pro Tyr Ser Phe Ala Asp Gly Ile
210 215 220
Asp Ser Ile Ser Val Ser Gly His Lys Met Ile Gly Ser Pro Met Pro
225 230 235 240
Cys Gly Ile Val Leu Ala Lys Arg Ser Asn Val Ser Arg Ile Ser Val
245 250 255
Glu Ile Asp Tyr Ile Ser Ala Lys Asp Gln Thr Ile Ser Gly Ser Arg
260 265 270
Asn Gly His Thr Pro Met Met Leu Trp Ala Ala Ile Lys Ser Arg Pro
275 280 285
Leu Ala Glu Trp Arg Arg Lys Val Arg His Cys Leu Asp Met Ala Gln
290 295 300
Tyr Ala Ile Asp Arg Leu Gln Ala Ala Gly Ile Gln Ala Trp Arg Cys
305 310 315 320
Lys Asn Ser Ile Thr Val Val Phe Pro Ser Pro Ser Glu Pro Val Cys
325 330 335
Asp Lys His Gly Leu Ala Arg Ser Gly Gly Thr Ala His Leu Ile Thr
340 345 350
Thr Pro His His His Asp Ser Gln Arg Leu Asp Arg Leu Leu Asp Asp
355 360 365
Ile Val Gln Asp Leu Gly Ala Met Thr Ala Pro Ala Gly Ala Thr Met
370 375 380
Ser Ala Ala
385
<210> 7
<211> 281
<212> PRT
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 7
Met Leu Lys Ile Ala Val Pro Asn Lys Gly Ser Leu Ser Glu Arg Ala
1 5 10 15
Met Glu Ile Leu Ala Glu Ala Gly Tyr Ala Gly Arg Gly Asp Ser Lys
20 25 30
Ser Leu Asn Val Phe Asp Glu Ala Asn Asn Val Glu Phe Phe Phe Leu
35 40 45
Arg Pro Lys Asp Ile Ala Ile Tyr Val Ala Gly Gly Gln Leu Asp Leu
50 55 60
Gly Ile Thr Gly Arg Asp Leu Ala Arg Asp Ser Gln Ala Asp Val His
65 70 75 80
Glu Val Leu Ser Leu Gly Phe Gly Ser Ser Thr Phe Arg Tyr Ala Ala
85 90 95
Pro Ala Asp Glu Glu Trp Ser Ile Glu Lys Leu Asp Gly Lys Arg Ile
100 105 110
Ala Thr Ser Tyr Pro Asn Leu Val Arg Asp Asp Leu Ala Ala Arg Gly
115 120 125
Leu Ser Ala Glu Val Leu Arg Leu Asp Gly Ala Val Glu Val Ser Ile
130 135 140
Lys Leu Gly Val Ala Asp Ala Ile Ala Asp Val Val Ser Thr Gly Arg
145 150 155 160
Thr Leu Arg Gln Gln Gly Leu Ala Pro Phe Gly Glu Val Leu Cys Thr
165 170 175
Ser Glu Ala Val Ile Val Gly Arg Lys Asp Glu Lys Val Thr Pro Glu
180 185 190
Gln Gln Ile Leu Leu Arg Arg Ile Gln Gly Ile Leu His Ala Gln Asn
195 200 205
Phe Leu Met Leu Asp Tyr Lys Val Asp Arg Asp Asn Leu Asp Ala Ala
210 215 220
Thr Ala Val Thr Pro Gly Phe Ser Gly Pro Ala Val Ser Pro Leu Ala
225 230 235 240
Arg Asp Asn Trp Val Ala Val Arg Ala Met Val Pro Arg Arg Ser Ala
245 250 255
Asn Ala Ile Met Asp Lys Leu Ala Gly Leu Gly Ala Glu Ala Ile Leu
260 265 270
Ala Ser Glu Ile Arg Ile Ala Arg Ile
275 280
<210> 8
<211> 334
<212> PRT
<213> 巴氏甲烷八叠球菌（Methanosarcina barkeri）菌株Wiesmoor
<400> 8
Met Gln Lys Tyr Glu Tyr Asp Pro Lys Trp Val Ile Asn Asn Ala Ile
1 5 10 15
Ser Ser Glu Arg Glu Phe Cys Thr Gly Tyr Gln Asn Pro Gly Ala Ser
20 25 30
Gly Asn Gly Tyr Val Thr Thr Ile Lys Leu Ser Thr Gly Leu Val Asp
35 40 45
Ile Thr Pro Trp Glu Lys Val Gln Ala Met Ser Glu His Glu Val Ile
50 55 60
Lys Phe Asp Arg Gly Cys Ser Asn Ile Val Ser Tyr Asp Arg Cys Glu
65 70 75 80
Cys Asn Asp Ala Tyr Ile Gly Ala Ile Asn Met Leu Thr Ala Ser Ser
85 90 95
Phe Ser Gly Leu Gln Gly Val Ile Trp Gly Tyr Asp Ile Ala Val Val
100 105 110
Glu Asn Leu Arg Ser Arg Lys Leu Tyr Asp Gln Lys Trp Pro Ser Gly
115 120 125
Asp Pro Asn Ser Glu Tyr Ser Thr Pro Val Tyr Ser Ile Glu Pro Leu
130 135 140
Leu Asn Ala Thr Glu Arg Leu Phe Gly His Ala Glu Pro Gly Lys Arg
145 150 155 160
Arg Phe Asn Pro Ile Pro Gly Ser Met Val Val Cys Ala Asn Lys Ser
165 170 175
Ala Thr Ser Asp Pro Ser Ser Asp Val Lys Glu Gly Trp Ala Phe Ser
180 185 190
Val Ile Ser Leu Ala Ile Leu Glu Asn Arg Asn Ser Gly Ser Asn Leu
195 200 205
Phe Ile Glu Asp Cys Asp Ile Ile Asp Ile Asn Asn Pro Asp Gly Thr
210 215 220
Arg Lys Thr Lys Glu Asp Val Lys Ala Met Leu Asp Thr Thr Leu Arg
225 230 235 240
Lys Val Thr Glu Cys Thr Val Leu Cys Gly Leu Asp Gln His Ile Lys
245 250 255
Tyr Lys Glu Ile Phe Ile Gly Tyr Lys Val Ile Lys Phe Asn Glu Lys
260 265 270
Gln Val Gly Cys Ala Leu Ala Cys Ala Pro Tyr Val Thr Leu Ala Arg
275 280 285
Asn Ala Val Pro Gln Gly Met Lys Pro Ser Lys Leu Thr Asp Met Asn
290 295 300
Ile Ser Gln Trp Glu Asn Ala Leu Asn Leu Gln Pro Leu Glu Lys Ile
305 310 315 320
Glu Lys Ser Lys Ile Gly Ile Leu Gly Met Gly Val Leu Asp
325 330
<210> 9
<211> 349
<212> PRT
<213> 巴氏甲烷八叠球菌（Methanosarcina barkeri）（菌株Fusaro / DSM 804）
<400> 9
Met Thr Leu His Ser Arg Tyr Ser Lys Leu Lys Gly Glu Asn Lys Met
1 5 10 15
Gln Lys Tyr Glu Tyr Asp Pro Lys Trp Val Ile Asn Asn Ala Ile Ser
20 25 30
Ser Glu Arg Glu Phe Cys Thr Gly Tyr Gln Asn Pro Gly Ala Ser Gly
35 40 45
Asn Gly Tyr Val Thr Thr Ile Lys Leu Ser Thr Gly Leu Val Asp Ile
50 55 60
Thr Pro Trp Glu Lys Val Gln Ala Met Ser Glu His Glu Val Ile Lys
65 70 75 80
Phe Asp Arg Gly Cys Ser Asn Ile Val Ser Tyr Asp Arg Cys Glu Cys
85 90 95
Asn Asp Ala Tyr Ile Gly Ala Ile Asn Met Leu Thr Ala Ser Ser Phe
100 105 110
Ser Gly Leu Gln Gly Val Ile Trp Gly Tyr Asp Ile Ala Val Val Glu
115 120 125
Asn Leu Arg Ser Arg Lys Leu Tyr Asp Gln Lys Trp Pro Ser Gly Asp
130 135 140
Pro Asn Ser Glu Tyr Ser Thr Pro Val Tyr Ser Ile Glu Pro Leu Leu
145 150 155 160
Asn Ala Thr Glu Arg Leu Phe Gly His Ala Glu Pro Gly Lys Arg Arg
165 170 175
Phe Asn Pro Ile Pro Gly Ser Met Val Val Cys Ala Asn Lys Ser Ala
180 185 190
Thr Ser Asp Pro Ser Ser Asp Val Lys Glu Gly Trp Ala Phe Ser Val
195 200 205
Ile Ser Leu Ala Ile Leu Glu Asn Arg Asn Ser Gly Ser Asn Leu Phe
210 215 220
Ile Glu Asp Cys Asp Ile Ile Asp Ile Asn Asn Pro Asp Gly Thr Arg
225 230 235 240
Lys Thr Lys Glu Asp Val Lys Ala Met Leu Asp Thr Thr Leu Arg Lys
245 250 255
Val Thr Glu Cys Thr Val Leu Cys Gly Leu Asp Gln His Ile Lys Tyr
260 265 270
Lys Glu Ile Phe Ile Gly Tyr Lys Val Ile Lys Phe Asn Glu Lys Gln
275 280 285
Val Gly Cys Ala Leu Ala Cys Ala Pro Tyr Val Thr Leu Ala Arg Asn
290 295 300
Ala Val Pro Gln Gly Met Lys Pro Ser Lys Leu Thr Asp Met Asn Ile
305 310 315 320
Ser Gln Trp Glu Asn Ala Leu Asn Leu Gln Pro Leu Glu Lys Ile Glu
325 330 335
Lys Ser Lys Ile Gly Ile Leu Gly Met Gly Val Leu Asp
340 345
<210> 10
<211> 847
<212> PRT
<213> 黑腹果蝇（Drosophila melanogaster）
<400> 10
Met Asp Phe Lys Glu Tyr Arg Gln Arg Gly Lys Glu Met Val Asp Tyr
1 5 10 15
Ile Ala Asp Tyr Leu Glu Asn Ile Arg Glu Arg Arg Val Phe Pro Asp
20 25 30
Val Ser Pro Gly Tyr Met Arg Gln Leu Leu Pro Glu Ser Ala Pro Ile
35 40 45
Glu Gly Glu Pro Trp Pro Lys Ile Phe Ser Asp Val Glu Arg Ile Val
50 55 60
Met Pro Gly Ile Thr His Trp Gln Ser Pro His Met His Ala Tyr Phe
65 70 75 80
Pro Ala Leu Asn Ser Met Pro Ser Leu Leu Gly Asp Met Leu Ala Asp
85 90 95
Ala Ile Asn Cys Leu Gly Phe Thr Trp Ala Ser Ser Pro Ala Cys Thr
100 105 110
Glu Leu Glu Ile Ile Val Met Asn Trp Leu Gly Lys Met Ile Gly Leu
115 120 125
Pro Asp Ala Phe Leu His Leu Ser Ser Gln Ser Gln Gly Gly Gly Val
130 135 140
Leu Gln Thr Thr Ala Ser Glu Ala Thr Leu Val Cys Leu Leu Ala Gly
145 150 155 160
Arg Thr Arg Ala Ile Gln Arg Phe His Glu Arg His Pro Gly Tyr Gln
165 170 175
Asp Ala Glu Ile Asn Ala Arg Leu Val Ala Tyr Cys Ser Asp Gln Ala
180 185 190
His Ser Ser Val Glu Lys Ala Ala Leu Ile Gly Leu Val Arg Met Arg
195 200 205
Tyr Ile Glu Ala Asp Asp Asp Leu Ala Met Arg Gly Lys Leu Leu Arg
210 215 220
Glu Ala Ile Glu Asp Asp Ile Lys Gln Gly Leu Val Pro Phe Trp Val
225 230 235 240
Cys Ala Thr Leu Gly Thr Thr Gly Ser Cys Ser Phe Asp Asn Leu Glu
245 250 255
Glu Ile Gly Ile Val Cys Ala Glu His His Leu Trp Leu His Val Asp
260 265 270
Ala Ala Tyr Ala Gly Ser Ala Phe Ile Cys Pro Glu Phe Arg Thr Trp
275 280 285
Leu Arg Gly Ile Glu Arg Ala Asp Ser Ile Ala Phe Asn Pro Ser Lys
290 295 300
Trp Leu Met Val His Phe Asp Ala Thr Ala Leu Trp Val Arg Asp Ser
305 310 315 320
Thr Ala Val His Arg Thr Phe Asn Val Glu Pro Leu Tyr Leu Gln His
325 330 335
Glu Asn Ser Gly Val Ala Val Asp Phe Met His Trp Gln Ile Pro Leu
340 345 350
Ser Arg Arg Phe Arg Ala Leu Lys Val Trp Phe Val Leu Arg Ser Tyr
355 360 365
Gly Ile Lys Gly Leu Gln Arg His Ile Arg Glu Gly Val Arg Leu Ala
370 375 380
Gln Lys Phe Glu Ala Leu Val Leu Ala Asp His Arg Phe Glu Leu Pro
385 390 395 400
Ala Lys Arg His Leu Gly Leu Val Val Phe Arg Ile Arg Gly Asp Asn
405 410 415
Glu Ile Thr Glu Lys Leu Leu Lys Arg Leu Asn His Arg Gly Asn Leu
420 425 430
His Cys Ile Pro Ser Ser Leu Lys Gly Gln Tyr Val Ile Arg Phe Thr
435 440 445
Ile Thr Ser Thr His Thr Thr Leu Asp Asp Ile Val Lys Asp Trp Met
450 455 460
Glu Ile Arg Gln Val Ala Ser Thr Val Leu Glu Glu Met Asn Ile Thr
465 470 475 480
Ile Ser Asn Arg Val Tyr Leu Lys Glu Thr Lys Glu Lys Asn Glu Ala
485 490 495
Phe Gly Ser Ser Leu Leu Leu Ser Asn Ser Pro Leu Ser Pro Lys Val
500 505 510
Val Asn Gly Ser Phe Ala Ala Ile Phe Asp Ala Asp Glu Phe Leu Ala
515 520 525
Lys Thr Tyr Ala Gly Val Arg Ile Ala His Gln Glu Ser Pro Ser Met
530 535 540
Arg Arg Arg Val Arg Gly Ile Leu Met Ser Gly Lys Gln Phe Ser Leu
545 550 555 560
Asp Ser His Met Asp Val Val Val Gln Thr Thr Leu Asp Ala Gly Asn
565 570 575
Gly Ala Thr Arg Thr Ser Thr Thr Asn Ser Tyr Gly His Thr Thr Ser
580 585 590
Ala Ala Gln Ala Asn Ser Glu Arg Gln Ala Ser Ile Gln Glu Asp Asn
595 600 605
Glu Glu Ser Pro Glu Glu Thr Glu Leu Leu Ser Leu Cys Arg Thr Ser
610 615 620
Asn Val Pro Ser Pro Glu His Ala His Ser Leu Ser Thr Pro Ser Arg
625 630 635 640
Ser Cys Ser Ser Ser Ser His Ser Leu Ile His Ser Leu Thr Gln Ser
645 650 655
Ser Pro Arg Ser Ser Pro Val Asn Gln Phe Arg Pro Ile Thr Leu Cys
660 665 670
Ala Val Pro Ser Gln Ser Gln Leu Ser Met Pro Leu Ala Met Pro Leu
675 680 685
Pro Asn Arg Asn Val Thr Val Ser Val Asp Ser Leu Leu Asn Pro Val
690 695 700
Thr Thr Cys Asn Val Tyr His Gly Lys Arg Phe Leu Glu Pro Leu Glu
705 710 715 720
Asn Leu Ala Gln Thr Ser Ala Ser Phe Ser Ser Ser Ile Phe Arg Leu
725 730 735
Pro Thr Pro Ile Ala Thr Pro Thr Arg Glu Ser Pro Glu Asp Pro Asp
740 745 750
Trp Pro Ala Lys Thr Phe Ser Gln Leu Leu Leu Glu Arg Tyr Ser Ser
755 760 765
Gln Ser Gln Ser Leu Gly Asn Asn Ser Ser Thr Glu Ser Ser Ser Leu
770 775 780
Ser Gly Gly Ala Thr Pro Thr Pro Thr Pro Met Ser Ser Leu Asp Glu
785 790 795 800
Leu Val Thr Pro Leu Leu Leu Ser Phe Ala Ser Pro Ser Gln Pro Met
805 810 815
Leu Ser Ala His Gly Ile Gly Glu Gly Gln Arg Glu Gln Gly Ser Asp
820 825 830
Ser Asp Ala Thr Val Cys Ser Thr Thr Ser Ser Met Glu Ser Leu
835 840 845
<210> 11
<211> 413
<212> PRT
<213> 番茄（Solanum lycopersicum）
<400> 11
Met Glu Ser Asp Ile Lys Asn Glu Thr Ser Phe Gln Glu Leu Asp Met
1 5 10 15
Ile Leu Thr Gln Tyr Leu Glu Thr Leu Ser Glu Arg Lys Lys Tyr His
20 25 30
Ile Gly Tyr Pro Ile Asn Met Cys Tyr Glu His His Ala Thr Leu Ala
35 40 45
Pro Leu Leu Gln Phe His Leu Asn Asn Cys Gly Asp Pro Phe Thr Gln
50 55 60
His Pro Thr Asp Phe His Ser Lys Asp Phe Glu Val Ala Val Leu Asp
65 70 75 80
Trp Phe Ala Gln Leu Trp Glu Ile Glu Lys Asp Glu Tyr Trp Gly Tyr
85 90 95
Ile Thr Ser Gly Gly Thr Glu Gly Asn Leu His Gly Phe Trp Leu Gly
100 105 110
Arg Arg Glu Leu Leu Pro Asn Gly Tyr Leu Tyr Ala Ser Lys Asp Ser
115 120 125
His Tyr Ser Ile Phe Lys Ala Ala Arg Met Tyr Arg Met Glu Leu Gln
130 135 140
Thr Ile Asn Thr Leu Val Asn Gly Glu Ile Asp Tyr Glu Asp Leu Gln
145 150 155 160
Ser Lys Leu Leu Val Asn Lys Asn Lys Pro Ala Ile Ile Asn Ile Asn
165 170 175
Ile Gly Thr Thr Phe Lys Gly Ala Ile Asp Asp Leu Asp Phe Val Ile
180 185 190
Gln Thr Leu Glu Asn Cys Gly Tyr Ser Asn Asp Asn Tyr Tyr Ile His
195 200 205
Cys Asp Arg Ala Leu Cys Gly Leu Ile Leu Pro Phe Ile Lys His Ala
210 215 220
Lys Lys Ile Thr Phe Lys Lys Pro Ile Gly Ser Ile Ser Ile Ser Gly
225 230 235 240
His Lys Phe Leu Gly Cys Pro Met Ser Cys Gly Val Gln Ile Thr Arg
245 250 255
Arg Ser Tyr Val Ser Thr Leu Ser Lys Ile Glu Tyr Ile Asn Ser Ala
260 265 270
Asp Ala Thr Ile Ser Gly Ser Arg Asn Gly Phe Thr Pro Ile Phe Leu
275 280 285
Trp Tyr Cys Leu Ser Lys Lys Gly His Ala Arg Leu Gln Gln Asp Ser
290 295 300
Ile Thr Cys Ile Glu Asn Ala Arg Tyr Leu Lys Asp Arg Leu Leu Glu
305 310 315 320
Ala Gly Ile Ser Val Met Leu Asn Asp Phe Ser Ile Thr Val Val Phe
325 330 335
Glu Arg Pro Cys Asp His Lys Phe Ile Arg Arg Trp Asn Leu Cys Cys
340 345 350
Leu Arg Gly Met Ala His Val Val Ile Met Pro Gly Ile Thr Arg Glu
355 360 365
Thr Ile Asp Ser Phe Phe Lys Asp Leu Met Gln Glu Arg Asn Tyr Lys
370 375 380
Trp Tyr Gln Asp Val Lys Ala Leu Pro Pro Cys Leu Ala Asp Asp Leu
385 390 395 400
Ala Leu Asn Cys Met Cys Ser Asn Lys Lys Met His Asn
405 410
<210> 12
<211> 662
<212> PRT
<213> 小家鼠（Mus musculus）
<400> 12
Met Met Glu Pro Cys Glu Tyr Arg Glu Tyr Arg Glu Tyr Tyr Arg Ala
1 5 10 15
Arg Gly Lys Glu Met Val Asp Tyr Ile Ser Gln Tyr Leu Ser Thr Val
20 25 30
Arg Glu Arg Gln Val Thr Pro Asn Val Gln Pro Gly Tyr Leu Arg Ala
35 40 45
Gln Leu Pro Ala Ser Ala Pro Glu Glu Pro Asp Ser Trp Asp Ser Ile
50 55 60
Phe Gly Asp Ile Glu Arg Val Ile Met Pro Gly Val Val His Trp Gln
65 70 75 80
Ser Pro His Met His Ala Tyr Tyr Pro Ala Leu Thr Ser Trp Pro Ser
85 90 95
Leu Leu Gly Asp Met Leu Ala Asp Ala Ile Asn Cys Leu Gly Phe Thr
100 105 110
Trp Ala Ser Ser Pro Ala Cys Thr Glu Leu Glu Met Asn Ile Met Asp
115 120 125
Trp Leu Ala Lys Met Leu Gly Leu Pro Glu Tyr Phe Leu His His His
130 135 140
Pro Ser Ser Arg Gly Gly Gly Val Leu Gln Ser Thr Val Ser Glu Ser
145 150 155 160
Thr Leu Ile Ala Leu Leu Ala Ala Arg Lys Asn Lys Ile Leu Ala Met
165 170 175
Lys Ala Cys Glu Pro Asp Ala Asn Glu Ser Ser Leu Asn Ala Arg Leu
180 185 190
Val Ala Tyr Thr Ser Asp Gln Ala His Ser Ser Val Glu Lys Ala Gly
195 200 205
Leu Ile Ser Leu Val Lys Ile Arg Phe Leu Pro Val Asp Asp Asn Phe
210 215 220
Ser Leu Arg Gly Glu Ala Leu Gln Lys Ala Ile Glu Glu Asp Lys Gln
225 230 235 240
Gln Gly Leu Val Pro Val Phe Val Cys Ala Thr Leu Gly Thr Thr Gly
245 250 255
Val Cys Ala Phe Asp Arg Leu Ser Glu Leu Gly Pro Ile Cys Ala Ser
260 265 270
Glu Gly Leu Trp Leu His Val Asp Ala Ala Tyr Ala Gly Thr Ala Phe
275 280 285
Leu Cys Pro Glu Leu Arg Gly Phe Leu Glu Gly Ile Glu Tyr Ala Asp
290 295 300
Ser Phe Thr Phe Asn Pro Ser Lys Trp Met Met Val His Phe Asp Cys
305 310 315 320
Thr Gly Phe Trp Val Lys Asp Lys Tyr Lys Leu Gln Gln Thr Phe Ser
325 330 335
Val Asn Pro Ile Tyr Leu Arg His Ala Asn Ser Gly Ala Ala Thr Asp
340 345 350
Phe Met His Trp Gln Ile Pro Leu Ser Arg Arg Phe Arg Ser Ile Lys
355 360 365
Leu Trp Phe Val Ile Arg Ser Phe Gly Val Lys Asn Leu Gln Ala His
370 375 380
Val Arg His Gly Thr Glu Met Ala Lys Tyr Phe Glu Ser Leu Val Arg
385 390 395 400
Ser Asp Pro Ser Phe Glu Ile Pro Ala Lys Arg His Leu Gly Leu Val
405 410 415
Val Phe Arg Leu Lys Gly Pro Asn Cys Leu Thr Glu Ser Val Leu Lys
420 425 430
Glu Ile Ala Lys Ala Gly Gln Leu Phe Leu Ile Pro Ala Thr Ile Gln
435 440 445
Asp Lys Leu Ile Ile Arg Phe Thr Val Thr Ser Gln Phe Thr Thr Lys
450 455 460
Glu Asp Ile Leu Arg Asp Trp His Leu Ile Gln Glu Ala Ala Asn Leu
465 470 475 480
Val Leu Ser Gln His Cys Thr Ser Gln Pro Ser Pro Arg Ala Lys Asn
485 490 495
Val Ile Pro Pro Pro Pro Gly Thr Arg Gly Leu Ser Leu Glu Ser Val
500 505 510
Ser Glu Gly Gly Asp Asp Pro Ala Gln Ala Arg Lys Ile Ile Lys Gln
515 520 525
Pro Gly Ala Ser Leu Ala Arg Arg Glu Gly Gly Ser Asp Leu Glu Thr
530 535 540
Met Pro Asp Pro Phe Asp Asp Cys Phe Ser Glu Glu Ala Pro Asn Thr
545 550 555 560
Thr Lys His Lys Leu Ser Ser Phe Leu Phe Ser Tyr Leu Ser Val Gln
565 570 575
Asn Arg Arg Lys Thr Thr Arg Ser Leu Ser Cys Asn Ser Val Pro Met
580 585 590
Ser Ala Gln Lys Ser Leu Pro Ala Asp Ala Ser Leu Lys Asn Gly Gly
595 600 605
Ser Phe Arg Ala Arg Ile Phe Ser Gly Phe Pro Glu Gln Met Met Met
610 615 620
Met Lys Lys Gly Ala Phe Lys Lys Leu Ile Lys Phe Tyr Ser Val Pro
625 630 635 640
Ser Phe Pro Glu Cys Ser Ser Gln Cys Ala Arg Gln Leu Pro Cys Cys
645 650 655
Pro Leu Glu Ala Met Val
660
<210> 13
<211> 246
<212> PRT
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 13
Met Thr Phe Thr Ile Leu Pro Ala Val Asp Val Val Asn Gly Gln Ala
1 5 10 15
Val Arg Leu Asp Gln Gly Glu Ala Gly Thr Glu Lys Ser Tyr Gly Thr
20 25 30
Pro Leu Glu Ser Ala Leu Lys Trp Gln Glu Gln Gly Ala Lys Trp Leu
35 40 45
His Phe Val Asp Leu Asp Ala Ala Phe Asn Arg Gly Ser Asn His Glu
50 55 60
Met Met Ala Glu Ile Val Gly Lys Leu Asp Val Asp Val Glu Leu Thr
65 70 75 80
Gly Gly Ile Arg Asp Asp Glu Ser Leu Glu Arg Ala Leu Ala Thr Gly
85 90 95
Ala Arg Arg Val Asn Ile Gly Thr Ala Ala Leu Glu Lys Pro Glu Trp
100 105 110
Ile Ala Ser Ala Ile Gln Arg Tyr Gly Glu Lys Ile Ala Val Asp Ile
115 120 125
Ala Val Arg Leu Glu Asp Gly Glu Trp Arg Thr Arg Gly Asn Gly Trp
130 135 140
Val Ser Asp Gly Gly Asp Leu Trp Asp Val Leu Glu Arg Leu Asp Ser
145 150 155 160
Gln Gly Cys Ala Arg Phe Val Val Thr Asp Val Ser Lys Asp Gly Thr
165 170 175
Leu Ser Gly Pro Asn Val Glu Leu Leu Arg Glu Val Ala Ala Ala Thr
180 185 190
Asp Ala Pro Ile Val Ala Ser Gly Gly Ile Ser Val Leu Glu Asp Val
195 200 205
Leu Glu Leu Ala Lys Tyr Gln Asp Glu Gly Ile Asp Ser Val Ile Ile
210 215 220
Gly Lys Ala Leu Tyr Glu His Lys Phe Thr Leu Glu Glu Ala Leu Ala
225 230 235 240
Ala Val Glu Lys Leu Gly
245
<210> 14
<211> 366
<212> PRT
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 14
Met Thr Lys Ile Thr Leu Ser Asp Leu Pro Leu Arg Glu Glu Leu Arg
1 5 10 15
Gly Glu His Ala Tyr Gly Ala Pro Gln Leu Asn Val Asp Ile Arg Leu
20 25 30
Asn Thr Asn Glu Asn Pro Tyr Pro Pro Ser Glu Ala Leu Val Ala Asp
35 40 45
Leu Val Ala Thr Val Asp Lys Ile Ala Thr Glu Leu Asn Arg Tyr Pro
50 55 60
Glu Arg Asp Ala Val Glu Leu Arg Asp Glu Leu Ala Ala Tyr Ile Thr
65 70 75 80
Lys Gln Thr Gly Val Ala Val Thr Arg Asp Asn Leu Trp Ala Ala Asn
85 90 95
Gly Ser Asn Glu Ile Leu Gln Gln Leu Leu Gln Ala Phe Gly Gly Pro
100 105 110
Gly Arg Thr Ala Leu Gly Phe Gln Pro Ser Tyr Ser Met His Pro Ile
115 120 125
Leu Ala Lys Gly Thr His Thr Glu Phe Ile Ala Val Ser Arg Gly Ala
130 135 140
Asp Phe Arg Ile Asp Met Asp Val Ala Leu Glu Glu Ile Arg Ala Lys
145 150 155 160
Gln Pro Asp Ile Val Phe Val Thr Thr Pro Asn Asn Pro Thr Gly Asp
165 170 175
Val Thr Ser Leu Asp Asp Ile Glu Arg Ile Ile Asn Val Ala Pro Gly
180 185 190
Ile Val Ile Val Asp Glu Ala Tyr Ala Glu Phe Ser Pro Ser Pro Ser
195 200 205
Ala Thr Thr Leu Leu Glu Lys Tyr Pro Thr Lys Leu Val Val Ser Arg
210 215 220
Thr Met Ser Lys Ala Phe Asp Phe Ala Gly Gly Arg Leu Gly Tyr Phe
225 230 235 240
Val Ala Asn Pro Ala Phe Ile Asp Ala Val Met Leu Val Arg Leu Pro
245 250 255
Tyr His Leu Ser Ala Leu Ser Gln Ala Ala Ala Ile Val Ala Leu Arg
260 265 270
His Ser Ala Asp Thr Leu Gly Thr Val Glu Lys Leu Ser Val Glu Arg
275 280 285
Val Arg Val Ala Ala Arg Leu Glu Glu Leu Gly Tyr Ala Val Val Pro
290 295 300
Ser Glu Ser Asn Phe Val Phe Phe Gly Asp Phe Ser Asp Gln His Ala
305 310 315 320
Ala Trp Gln Ala Phe Leu Asp Arg Gly Val Leu Ile Arg Asp Val Gly
325 330 335
Ile Ala Gly His Leu Arg Thr Thr Ile Gly Val Pro Glu Glu Asn Asp
340 345 350
Ala Phe Leu Asp Ala Ala Ala Glu Ile Ile Lys Leu Asn Leu
355 360 365
<210> 15
<211> 442
<212> PRT
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 15
Met Leu Asn Val Thr Asp Leu Arg Gly Gln Thr Pro Ser Lys Ser Asp
1 5 10 15
Ile Arg Arg Ala Leu Pro Arg Gly Gly Thr Asp Val Trp Ser Val Leu
20 25 30
Pro Ile Val Gln Pro Val Val Glu Asp Val Gln Asn Arg Gly Ala Glu
35 40 45
Ala Ala Leu Asp Tyr Gly Glu Lys Phe Asp His Ile Arg Pro Ala Ser
50 55 60
Val Arg Val Pro Ala Glu Val Ile Ala Ala Ala Glu Asn Thr Leu Asp
65 70 75 80
Pro Leu Val Arg Glu Ser Ile Glu Glu Ser Ile Arg Arg Val Arg Lys
85 90 95
Val His Ala Glu Gln Lys Pro Ala Glu His Thr Thr Glu Leu Ser Pro
100 105 110
Gly Gly Thr Val Thr Glu Arg Phe Met Pro Ile Asp Arg Val Gly Leu
115 120 125
Tyr Val Pro Gly Gly Asn Ala Val Tyr Pro Ser Ser Val Ile Met Asn
130 135 140
Thr Val Pro Ala Gln Glu Ala Gly Val Asn Ser Leu Val Val Ala Ser
145 150 155 160
Pro Pro Gln Ala Glu His Gly Gly Trp Pro His Pro Thr Ile Leu Ala
165 170 175
Ala Cys Ser Ile Leu Gly Val Asp Glu Val Trp Ala Val Gly Gly Gly
180 185 190
Gln Ala Val Ala Leu Leu Ala Tyr Gly Asp Asp Ala Ala Gly Leu Glu
195 200 205
Pro Val Asp Met Ile Thr Gly Pro Gly Asn Ile Phe Val Thr Ala Ala
210 215 220
Lys Arg Leu Val Arg Gly Val Val Gly Thr Asp Ser Glu Ala Gly Pro
225 230 235 240
Thr Glu Ile Ala Val Leu Ala Asp Ala Ser Ala Asn Ala Val Asn Val
245 250 255
Ala Tyr Asp Leu Ile Ser Gln Ala Glu His Asp Val Met Ala Ala Ser
260 265 270
Val Leu Ile Thr Asp Ser Glu Gln Leu Ala Lys Asp Val Asn Arg Glu
275 280 285
Ile Glu Ala Arg Tyr Ser Ile Thr Arg Asn Ala Glu Arg Val Ala Glu
290 295 300
Ala Leu Arg Gly Ala Gln Ser Gly Ile Val Leu Val Asp Asp Ile Ser
305 310 315 320
Val Gly Ile Gln Val Ala Asp Gln Tyr Ala Ala Glu His Leu Glu Ile
325 330 335
His Thr Glu Asn Ala Arg Ala Val Ala Glu Gln Ile Thr Asn Ala Gly
340 345 350
Ala Ile Phe Val Gly Asp Phe Ser Pro Val Pro Leu Gly Asp Tyr Ser
355 360 365
Ala Gly Ser Asn His Val Leu Pro Thr Ser Gly Ser Ala Arg Phe Ser
370 375 380
Ala Gly Leu Ser Thr His Met Phe Leu Arg Pro Val Asn Leu Ile Glu
385 390 395 400
Tyr Asp Glu Ala Ala Leu Lys Asp Val Ser Gln Val Val Ile Asn Phe
405 410 415
Ala Asn Ala Glu Asp Leu Pro Ala His Gly Glu Ala Ile Arg Ala Arg
420 425 430
Phe Glu Asn Leu Pro Thr Thr Asp Glu Ala
435 440
<210> 16
<211> 87
<212> PRT
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 16
Val Lys Thr Phe Asp Ser Leu Tyr Glu Glu Leu Leu Asn Arg Ala Gln
1 5 10 15
Thr Arg Pro Glu Gly Ser Gly Thr Val Ala Ala Leu Asp Lys Gly Ile
20 25 30
His His Leu Gly Lys Lys Val Ile Glu Glu Ala Gly Glu Val Trp Ile
35 40 45
Ala Ala Glu Tyr Glu Thr Asp Glu Glu Leu Ala Gly Glu Ile Ser Gln
50 55 60
Leu Ile Tyr Trp Thr Gln Val Ile Met Val Ala Arg Gly Leu Lys Pro
65 70 75 80
Glu Asp Ile Tyr Lys Asn Leu
85
<210> 17
<211> 258
<212> PRT
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 17
Met Gly Val Ala Ile Arg Val Ile Pro Cys Leu Asp Val Asp Asn Gly
1 5 10 15
Arg Val Val Lys Gly Val Asn Phe Glu Asn Leu Arg Asp Ala Gly Asp
20 25 30
Pro Val Glu Leu Ala Lys Arg Tyr Asp Glu Glu Gly Ala Asp Glu Leu
35 40 45
Thr Phe Leu Asp Val Thr Ala Ser Lys His Gly Arg Gly Thr Met Leu
50 55 60
Asp Val Val Arg Arg Thr Ala Asp Gln Val Phe Ile Pro Leu Thr Val
65 70 75 80
Gly Gly Gly Val Arg Ser Glu Glu Asp Val Asp Gln Leu Leu Arg Ala
85 90 95
Gly Ala Asp Lys Val Ser Val Asn Thr Ser Ala Ile Ala Arg Pro Glu
100 105 110
Leu Leu Ser Glu Leu Ser Lys Arg Phe Gly Ala Gln Cys Ile Val Leu
115 120 125
Ser Val Asp Ala Arg Arg Val Pro Glu Gly Gly Thr Pro Gln Pro Ser
130 135 140
Gly Phe Glu Val Thr Thr His Gly Gly Ser Lys Ser Ala Glu Leu Asp
145 150 155 160
Ala Ile Glu Trp Ala Lys Arg Gly Glu Glu Leu Gly Val Gly Glu Ile
165 170 175
Leu Leu Asn Ser Met Asp Gly Asp Gly Thr Lys Asn Gly Phe Asp Leu
180 185 190
Glu Leu Leu Glu Lys Val Arg Ala Ala Val Ser Ile Pro Val Ile Ala
195 200 205
Ser Gly Gly Ala Gly Lys Ala Glu His Phe Pro Pro Ala Val Ala Ala
210 215 220
Gly Ala Asn Ala Val Leu Ala Ala Thr Ile Phe His Phe Arg Glu Val
225 230 235 240
Thr Ile Ala Glu Val Lys Gly Ala Ile Lys Asp Ala Gly Phe Glu Val
245 250 255
Arg Lys
<210> 18
<211> 211
<212> PRT
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 18
Met Thr Lys Thr Val Ala Leu Leu Asp Tyr Gly Ser Gly Asn Leu Arg
1 5 10 15
Ser Ala Gln Arg Ala Leu Glu Arg Ala Gly Ala Glu Val Thr Val Ser
20 25 30
Ser Asp Pro Glu Val Cys Thr Asn Ala Asp Gly Leu Leu Val Pro Gly
35 40 45
Val Gly Ala Phe Asp Ala Cys Met Lys Gly Leu Lys Asn Val Phe Gly
50 55 60
His Arg Ile Ile Gly Gln Arg Leu Ala Gly Gly Arg Pro Val Met Gly
65 70 75 80
Ile Cys Val Gly Met Gln Ile Leu Phe Asp Glu Gly Asp Glu His Gly
85 90 95
Ile Lys Ser Ala Gly Cys Gly Glu Trp Pro Gly Lys Val Glu Arg Leu
100 105 110
Gln Ala Glu Ile Leu Pro His Met Gly Trp Asn Thr Leu Glu Met Pro
115 120 125
Thr Asn Ser Pro Met Phe Glu Gly Ile Ser Pro Asp Glu Arg Phe Tyr
130 135 140
Phe Val His Ser Tyr Gly Val Arg Lys Trp Thr Leu Glu Thr Asp Asp
145 150 155 160
Leu Thr Thr Pro Pro Glu Val Val Trp Ala Lys His Glu Asn Asp Arg
165 170 175
Phe Val Ala Ala Val Glu Asn Gly Thr Leu Trp Ala Thr Gln Phe His
180 185 190
Pro Glu Lys Ser Gly Asp Val Gly Ala Lys Leu Leu Arg Asn Trp Ile
195 200 205
Asn Tyr Ile
210
<210> 19
<211> 118
<212> PRT
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 19
Met Ser Asp Asn Pro Gln Glu Tyr Glu Leu Asp Trp Asp Val Glu Lys
1 5 10 15
Arg Leu Lys Leu Asn Asp Ala Gly Leu Val Pro Ala Ile Val Gln Ala
20 25 30
Asp Gly Thr Asn Glu Val Leu Met Met Ala Trp Met Asp Thr His Ala
35 40 45
Leu Ala Tyr Thr Leu Ala Thr Arg Arg Gly Thr Tyr Phe Ser Arg Ser
50 55 60
Arg Asn Glu Tyr Trp Ile Lys Gly Leu Thr Ser Gly Asn Val Gln Glu
65 70 75 80
Val Thr Gly Leu Ala Leu Asp Cys Asp Gly Asp Thr Val Leu Leu Thr
85 90 95
Val Lys Gln Thr Gly Gly Ala Cys His Thr Gly Ala His Thr Cys Phe
100 105 110
Asp Asn Asp Val Leu Leu
115
<210> 20
<211> 799
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 20
Met Val Leu Pro Ile Leu Pro Leu Ile Asp Asp Leu Ala Ser Trp Asn
1 5 10 15
Ser Lys Lys Glu Tyr Val Ser Leu Val Gly Gln Val Leu Leu Asp Gly
20 25 30
Ser Ser Leu Ser Asn Glu Glu Ile Leu Gln Phe Ser Lys Glu Glu Glu
35 40 45
Val Pro Leu Val Ala Leu Ser Leu Pro Ser Gly Lys Phe Ser Asp Asp
50 55 60
Glu Ile Ile Ala Phe Leu Asn Asn Gly Val Ser Ser Leu Phe Ile Ala
65 70 75 80
Ser Gln Asp Ala Lys Thr Ala Glu His Leu Val Glu Gln Leu Asn Val
85 90 95
Pro Lys Glu Arg Val Val Val Glu Glu Asn Gly Val Phe Ser Asn Gln
100 105 110
Phe Met Val Lys Gln Lys Phe Ser Gln Asp Lys Ile Val Ser Ile Lys
115 120 125
Lys Leu Ser Lys Asp Met Leu Thr Lys Glu Val Leu Gly Glu Val Arg
130 135 140
Thr Asp Arg Pro Asp Gly Leu Tyr Thr Thr Leu Val Val Asp Gln Tyr
145 150 155 160
Glu Arg Cys Leu Gly Leu Val Tyr Ser Ser Lys Lys Ser Ile Ala Lys
165 170 175
Ala Ile Asp Leu Gly Arg Gly Val Tyr Tyr Ser Arg Ser Arg Asn Glu
180 185 190
Ile Trp Ile Lys Gly Glu Thr Ser Gly Asn Gly Gln Lys Leu Leu Gln
195 200 205
Ile Ser Thr Asp Cys Asp Ser Asp Ala Leu Lys Phe Ile Val Glu Gln
210 215 220
Glu Asn Val Gly Phe Cys His Leu Glu Thr Met Ser Cys Phe Gly Glu
225 230 235 240
Phe Lys His Gly Leu Val Gly Leu Glu Ser Leu Leu Lys Gln Arg Leu
245 250 255
Gln Asp Ala Pro Glu Glu Ser Tyr Thr Arg Arg Leu Phe Asn Asp Ser
260 265 270
Ala Leu Leu Asp Ala Lys Ile Lys Glu Glu Ala Glu Glu Leu Thr Glu
275 280 285
Ala Lys Gly Lys Lys Glu Leu Ser Trp Glu Ala Ala Asp Leu Phe Tyr
290 295 300
Phe Ala Leu Ala Lys Leu Val Ala Asn Asp Val Ser Leu Lys Asp Val
305 310 315 320
Glu Asn Asn Leu Asn Met Lys His Leu Lys Val Thr Arg Arg Lys Gly
325 330 335
Asp Ala Lys Pro Lys Phe Val Gly Gln Pro Lys Ala Glu Glu Glu Lys
340 345 350
Leu Thr Gly Pro Ile His Leu Asp Val Val Lys Ala Ser Asp Lys Val
355 360 365
Gly Val Gln Lys Ala Leu Ser Arg Pro Ile Gln Lys Thr Ser Glu Ile
370 375 380
Met His Leu Val Asn Pro Ile Ile Glu Asn Val Arg Asp Lys Gly Asn
385 390 395 400
Ser Ala Leu Leu Glu Tyr Thr Glu Lys Phe Asp Gly Val Lys Leu Ser
405 410 415
Asn Pro Val Leu Asn Ala Pro Phe Pro Glu Glu Tyr Phe Glu Gly Leu
420 425 430
Thr Glu Glu Met Lys Glu Ala Leu Asp Leu Ser Ile Glu Asn Val Arg
435 440 445
Lys Phe His Ala Ala Gln Leu Pro Thr Glu Thr Leu Glu Val Glu Thr
450 455 460
Gln Pro Gly Val Leu Cys Ser Arg Phe Pro Arg Pro Ile Glu Lys Val
465 470 475 480
Gly Leu Tyr Ile Pro Gly Gly Thr Ala Ile Leu Pro Ser Thr Ala Leu
485 490 495
Met Leu Gly Val Pro Ala Gln Val Ala Gln Cys Lys Glu Ile Val Phe
500 505 510
Ala Ser Pro Pro Arg Lys Ser Asp Gly Lys Val Ser Pro Glu Val Val
515 520 525
Tyr Val Ala Glu Lys Val Gly Ala Ser Lys Ile Val Leu Ala Gly Gly
530 535 540
Ala Gln Ala Val Ala Ala Met Ala Tyr Gly Thr Glu Thr Ile Pro Lys
545 550 555 560
Val Asp Lys Ile Leu Gly Pro Gly Asn Gln Phe Val Thr Ala Ala Lys
565 570 575
Met Tyr Val Gln Asn Asp Thr Gln Ala Leu Cys Ser Ile Asp Met Pro
580 585 590
Ala Gly Pro Ser Glu Val Leu Val Ile Ala Asp Glu Asp Ala Asp Val
595 600 605
Asp Phe Val Ala Ser Asp Leu Leu Ser Gln Ala Glu His Gly Ile Asp
610 615 620
Ser Gln Val Ile Leu Val Gly Val Asn Leu Ser Glu Lys Lys Ile Gln
625 630 635 640
Glu Ile Gln Asp Ala Val His Asn Gln Ala Leu Gln Leu Pro Arg Val
645 650 655
Asp Ile Val Arg Lys Cys Ile Ala His Ser Thr Ile Val Leu Cys Asp
660 665 670
Gly Tyr Glu Glu Ala Leu Glu Met Ser Asn Gln Tyr Ala Pro Glu His
675 680 685
Leu Ile Leu Gln Ile Ala Asn Ala Asn Asp Tyr Val Lys Leu Val Asp
690 695 700
Asn Ala Gly Ser Val Phe Val Gly Ala Tyr Thr Pro Glu Ser Cys Gly
705 710 715 720
Asp Tyr Ser Ser Gly Thr Asn His Thr Leu Pro Thr Tyr Gly Tyr Ala
725 730 735
Arg Gln Tyr Ser Gly Ala Asn Thr Ala Thr Phe Gln Lys Phe Ile Thr
740 745 750
Ala Gln Asn Ile Thr Pro Glu Gly Leu Glu Asn Ile Gly Arg Ala Val
755 760 765
Met Cys Val Ala Lys Lys Glu Gly Leu Asp Gly His Arg Asn Ala Val
770 775 780
Lys Ile Arg Met Ser Lys Leu Gly Leu Ile Pro Lys Asp Phe Gln
785 790 795
<210> 21
<211> 552
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 21
Met Pro Val Val His Val Ile Asp Val Glu Ser Gly Asn Leu Gln Ser
1 5 10 15
Leu Thr Asn Ala Ile Glu His Leu Gly Tyr Glu Val Gln Leu Val Lys
20 25 30
Ser Pro Lys Asp Phe Asn Ile Ser Gly Thr Ser Arg Leu Ile Leu Pro
35 40 45
Gly Val Gly Asn Tyr Gly His Phe Val Asp Asn Leu Phe Asn Arg Gly
50 55 60
Phe Glu Lys Pro Ile Arg Glu Tyr Ile Glu Ser Gly Lys Pro Ile Met
65 70 75 80
Gly Ile Cys Val Gly Leu Gln Ala Leu Phe Ala Gly Ser Val Glu Ser
85 90 95
Pro Lys Ser Thr Gly Leu Asn Tyr Ile Asp Phe Lys Leu Ser Arg Phe
100 105 110
Asp Asp Ser Glu Lys Pro Val Pro Glu Ile Gly Trp Asn Ser Cys Ile
115 120 125
Pro Ser Glu Asn Leu Phe Phe Gly Leu Asp Pro Tyr Lys Arg Tyr Tyr
130 135 140
Phe Val His Ser Phe Ala Ala Ile Leu Asn Ser Glu Lys Lys Lys Asn
145 150 155 160
Leu Glu Asn Asp Gly Trp Lys Ile Ala Lys Ala Lys Tyr Gly Ser Glu
165 170 175
Glu Phe Ile Ala Ala Val Asn Lys Asn Asn Ile Phe Ala Thr Gln Phe
180 185 190
His Pro Glu Lys Ser Gly Lys Ala Gly Leu Asn Val Ile Glu Asn Phe
195 200 205
Leu Lys Gln Gln Ser Pro Pro Ile Pro Asn Tyr Ser Ala Glu Glu Lys
210 215 220
Glu Leu Leu Met Asn Asp Tyr Ser Asn Tyr Gly Leu Thr Arg Arg Ile
225 230 235 240
Ile Ala Cys Leu Asp Val Arg Thr Asn Asp Gln Gly Asp Leu Val Val
245 250 255
Thr Lys Gly Asp Gln Tyr Asp Val Arg Glu Lys Ser Asp Gly Lys Gly
260 265 270
Val Arg Asn Leu Gly Lys Pro Val Gln Leu Ala Gln Lys Tyr Tyr Gln
275 280 285
Gln Gly Ala Asp Glu Val Thr Phe Leu Asn Ile Thr Ser Phe Arg Asp
290 295 300
Cys Pro Leu Lys Asp Thr Pro Met Leu Glu Val Leu Lys Gln Ala Ala
305 310 315 320
Lys Thr Val Phe Val Pro Leu Thr Val Gly Gly Gly Ile Lys Asp Ile
325 330 335
Val Asp Val Asp Gly Thr Lys Ile Pro Ala Leu Glu Val Ala Ser Leu
340 345 350
Tyr Phe Arg Ser Gly Ala Asp Lys Val Ser Ile Gly Thr Asp Ala Val
355 360 365
Tyr Ala Ala Glu Lys Tyr Tyr Glu Leu Gly Asn Arg Gly Asp Gly Thr
370 375 380
Ser Pro Ile Glu Thr Ile Ser Lys Ala Tyr Gly Ala Gln Ala Val Val
385 390 395 400
Ile Ser Val Asp Pro Lys Arg Val Tyr Val Asn Ser Gln Ala Asp Thr
405 410 415
Lys Asn Lys Val Phe Glu Thr Glu Tyr Pro Gly Pro Asn Gly Glu Lys
420 425 430
Tyr Cys Trp Tyr Gln Cys Thr Ile Lys Gly Gly Arg Glu Ser Arg Asp
435 440 445
Leu Gly Val Trp Glu Leu Thr Arg Ala Cys Glu Ala Leu Gly Ala Gly
450 455 460
Glu Ile Leu Leu Asn Cys Ile Asp Lys Asp Gly Ser Asn Ser Gly Tyr
465 470 475 480
Asp Leu Glu Leu Ile Glu His Val Lys Asp Ala Val Lys Ile Pro Val
485 490 495
Ile Ala Ser Ser Gly Ala Gly Val Pro Glu His Phe Glu Glu Ala Phe
500 505 510
Leu Lys Thr Arg Ala Asp Ala Cys Leu Gly Ala Gly Met Phe His Arg
515 520 525
Gly Glu Phe Thr Val Asn Asp Val Lys Glu Tyr Leu Leu Glu His Gly
530 535 540
Leu Lys Val Arg Met Asp Glu Glu
545 550
<210> 22
<211> 281
<212> PRT
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）（菌株ATCC 13032 / DSM20300 / JCM 1318 / LMG 3730 / NCIMB 10025）
<400> 22
Met Leu Lys Ile Ala Val Pro Asn Lys Gly Ser Leu Ser Glu Arg Ala
1 5 10 15
Met Glu Ile Leu Ala Glu Ala Gly Tyr Ala Gly Arg Gly Asp Ser Lys
20 25 30
Ser Leu Asn Val Phe Asp Glu Ala Asn Asn Val Glu Phe Phe Phe Leu
35 40 45
Arg Pro Lys Asp Ile Ala Ile Tyr Val Ala Gly Gly Gln Leu Asp Leu
50 55 60
Gly Ile Thr Gly Arg Asp Leu Ala Arg Asp Ser Gln Ala Asp Val His
65 70 75 80
Glu Val Leu Ser Leu Gly Phe Gly Ser Ser Thr Phe Arg Tyr Ala Ala
85 90 95
Pro Ala Asp Glu Glu Trp Ser Ile Glu Lys Leu Asp Gly Lys Arg Ile
100 105 110
Ala Thr Ser Tyr Pro Asn Leu Val Arg Asp Asp Leu Ala Ala Arg Gly
115 120 125
Leu Ser Ala Glu Val Leu Arg Leu Asp Gly Ala Val Glu Val Ser Ile
130 135 140
Lys Leu Gly Val Ala Asp Ala Ile Ala Asp Val Val Ser Thr Gly Arg
145 150 155 160
Thr Leu Arg Gln Gln Gly Leu Ala Pro Phe Gly Glu Val Leu Cys Thr
165 170 175
Ser Glu Ala Val Ile Val Gly Arg Lys Asp Glu Lys Val Thr Pro Glu
180 185 190
Gln Gln Ile Leu Leu Arg Arg Ile Gln Gly Ile Leu His Ala Gln Asn
195 200 205
Phe Leu Met Leu Asp Tyr Lys Val Asp Arg Asp Asn Leu Asp Ala Ala
210 215 220
Thr Ala Val Thr Pro Gly Phe Ser Gly Pro Ala Val Ser Pro Leu Ala
225 230 235 240
Arg Asp Asn Trp Val Ala Val Arg Ala Met Val Pro Arg Arg Ser Ala
245 250 255
Asn Ala Ile Met Asp Lys Leu Ala Gly Leu Gly Ala Glu Ala Ile Leu
260 265 270
Ala Ser Glu Ile Arg Ile Ala Arg Ile
275 280
<210> 23
<211> 385
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 23
Met Val Phe Asp Leu Lys Arg Ile Val Arg Pro Lys Ile Tyr Asn Leu
1 5 10 15
Glu Pro Tyr Arg Cys Ala Arg Asp Asp Phe Thr Glu Gly Ile Leu Leu
20 25 30
Asp Ala Asn Glu Asn Ala His Gly Pro Thr Pro Val Glu Leu Ser Lys
35 40 45
Thr Asn Leu His Arg Tyr Pro Asp Pro His Gln Leu Glu Phe Lys Thr
50 55 60
Ala Met Thr Lys Tyr Arg Asn Lys Thr Ser Ser Tyr Ala Asn Asp Pro
65 70 75 80
Glu Val Lys Pro Leu Thr Ala Asp Asn Leu Cys Leu Gly Val Gly Ser
85 90 95
Asp Glu Ser Ile Asp Ala Ile Ile Arg Ala Cys Cys Val Pro Gly Lys
100 105 110
Glu Lys Ile Leu Val Leu Pro Pro Thr Tyr Ser Met Tyr Ser Val Cys
115 120 125
Ala Asn Ile Asn Asp Ile Glu Val Val Gln Cys Pro Leu Thr Val Ser
130 135 140
Asp Gly Ser Phe Gln Met Asp Thr Glu Ala Val Leu Thr Ile Leu Lys
145 150 155 160
Asn Asp Ser Leu Ile Lys Leu Met Phe Val Thr Ser Pro Gly Asn Pro
165 170 175
Thr Gly Ala Lys Ile Lys Thr Ser Leu Ile Glu Lys Val Leu Gln Asn
180 185 190
Trp Asp Asn Gly Leu Val Val Val Asp Glu Ala Tyr Val Asp Phe Cys
195 200 205
Gly Gly Ser Thr Ala Pro Leu Val Thr Lys Tyr Pro Asn Leu Val Thr
210 215 220
Leu Gln Thr Leu Ser Lys Ser Phe Gly Leu Ala Gly Ile Arg Leu Gly
225 230 235 240
Met Thr Tyr Ala Thr Ala Glu Leu Ala Arg Ile Leu Asn Ala Met Lys
245 250 255
Ala Pro Tyr Asn Ile Ser Ser Leu Ala Ser Glu Tyr Ala Leu Lys Ala
260 265 270
Val Gln Asp Ser Asn Leu Lys Lys Met Glu Ala Thr Ser Lys Ile Ile
275 280 285
Asn Glu Glu Lys Met Arg Leu Leu Lys Glu Leu Thr Ala Leu Asp Tyr
290 295 300
Val Asp Asp Gln Tyr Val Gly Gly Leu Asp Ala Asn Phe Leu Leu Ile
305 310 315 320
Arg Ile Asn Gly Gly Asp Asn Val Leu Ala Lys Lys Leu Tyr Tyr Gln
325 330 335
Leu Ala Thr Gln Ser Gly Val Val Val Arg Phe Arg Gly Asn Glu Leu
340 345 350
Gly Cys Ser Gly Cys Leu Arg Ile Thr Val Gly Thr His Glu Glu Asn
355 360 365
Thr His Leu Ile Lys Tyr Phe Lys Glu Thr Leu Tyr Lys Leu Ala Asn
370 375 380
Glu
385
<210> 24
<211> 220
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 24
Met Thr Glu Gln Lys Ala Leu Val Lys Arg Ile Thr Asn Glu Thr Lys
1 5 10 15
Ile Gln Ile Ala Ile Ser Leu Lys Gly Gly Pro Leu Ala Ile Glu His
20 25 30
Ser Ile Phe Pro Glu Lys Glu Ala Glu Ala Val Ala Glu Gln Ala Thr
35 40 45
Gln Ser Gln Val Ile Asn Val His Thr Gly Ile Gly Phe Leu Asp His
50 55 60
Met Ile His Ala Leu Ala Lys His Ser Gly Trp Ser Leu Ile Val Glu
65 70 75 80
Cys Ile Gly Asp Leu His Ile Asp Asp His His Thr Thr Glu Asp Cys
85 90 95
Gly Ile Ala Leu Gly Gln Ala Phe Lys Glu Ala Leu Gly Ala Val Arg
100 105 110
Gly Val Lys Arg Phe Gly Ser Gly Phe Ala Pro Leu Asp Glu Ala Leu
115 120 125
Ser Arg Ala Val Val Asp Leu Ser Asn Arg Pro Tyr Ala Val Val Glu
130 135 140
Leu Gly Leu Gln Arg Glu Lys Val Gly Asp Leu Ser Cys Glu Met Ile
145 150 155 160
Pro His Phe Leu Glu Ser Phe Ala Glu Ala Ser Arg Ile Thr Leu His
165 170 175
Val Asp Cys Leu Arg Gly Lys Asn Asp His His Arg Ser Glu Ser Ala
180 185 190
Phe Lys Ala Leu Ala Val Ala Ile Arg Glu Ala Thr Ser Pro Asn Gly
195 200 205
Thr Asn Asp Val Pro Ser Thr Lys Gly Val Leu Met
210 215 220
<210> 25
<211> 335
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 25
Met His Ser His His Ser His Ser Gly Asp Tyr Ser Ala His Gly Thr
1 5 10 15
Asp Pro Leu Asp Ser Val Val Asp Gln Val Val Asn Leu Asn Phe His
20 25 30
Thr Tyr Cys Leu Thr Glu His Ile Pro Arg Ile Glu Ala Lys Phe Ile
35 40 45
Tyr Pro Glu Glu Gln Ser Leu Gly Lys Asn Pro Glu Glu Val Ile Ser
50 55 60
Lys Leu Glu Thr Ser Phe Lys Asn Phe Met Ser His Ala Gln Glu Ile
65 70 75 80
Lys Thr Arg Tyr Ala Asp Arg Pro Asp Val Arg Thr Lys Phe Ile Ile
85 90 95
Gly Met Glu Ile Glu Ser Cys Asp Met Ala His Ile Glu Tyr Ala Lys
100 105 110
Arg Leu Met Lys Glu Asn Asn Asp Thr Leu Lys Phe Cys Val Gly Ser
115 120 125
Val His His Val Asn Gly Ile Pro Ile Asp Phe Asp Gln Gln Gln Trp
130 135 140
Tyr Asn Ser Leu His Ser Phe Asn Asp Asn Leu Lys Asp Phe Leu Leu
145 150 155 160
Ser Tyr Phe Gln Ser Gln Tyr Glu Met Leu Ile Asn Ile Lys Pro Leu
165 170 175
Val Val Gly His Phe Asp Leu Tyr Lys Leu Phe Leu Pro Asn Asp Met
180 185 190
Leu Val Asn Gln Lys Ser Gly Asn Cys Asn Glu Glu Thr Gly Val Pro
195 200 205
Val Ala Ser Leu Asp Val Ile Ser Glu Trp Pro Glu Ile Tyr Asp Ala
210 215 220
Val Val Arg Asn Leu Gln Phe Ile Asp Ser Tyr Gly Gly Ala Ile Glu
225 230 235 240
Ile Asn Thr Ser Ala Leu Arg Lys Gly Leu Glu Glu Pro Tyr Pro Ser
245 250 255
Lys Thr Leu Cys Asn Leu Val Lys Lys His Cys Gly Ser Arg Phe Val
260 265 270
Leu Ser Asp Asp Ala His Gly Val Ala Gln Val Gly Val Cys Tyr Asp
275 280 285
Lys Val Lys Lys Tyr Ile Val Asp Val Leu Gln Leu Glu Tyr Ile Cys
290 295 300
Tyr Leu Glu Glu Ser Gln Ser Pro Glu Asn Val Leu Thr Val Lys Arg
305 310 315 320
Leu Pro Ile Ser Gln Phe Val Asn Asp Pro Phe Trp Ala Asn Ile
325 330 335
<210> 26
<211> 514
<212> PRT
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 26
Met Ser Thr Asn Thr Thr Pro Ser Ser Trp Thr Asn Pro Leu Arg Asp
1 5 10 15
Pro Gln Asp Lys Arg Leu Pro Arg Ile Ala Gly Pro Ser Gly Met Val
20 25 30
Ile Phe Gly Val Thr Gly Asp Leu Ala Arg Lys Lys Leu Leu Pro Ala
35 40 45
Ile Tyr Asp Leu Ala Asn Arg Gly Leu Leu Pro Pro Gly Phe Ser Leu
50 55 60
Val Gly Tyr Gly Arg Arg Glu Trp Ser Lys Glu Asp Phe Glu Lys Tyr
65 70 75 80
Val Arg Asp Ala Ala Ser Ala Gly Ala Arg Thr Glu Phe Arg Glu Asn
85 90 95
Val Trp Glu Arg Leu Ala Glu Gly Met Glu Phe Val Arg Gly Asn Phe
100 105 110
Asp Asp Asp Ala Ala Phe Asp Asn Leu Ala Ala Thr Leu Lys Arg Ile
115 120 125
Asp Lys Thr Arg Gly Thr Ala Gly Asn Trp Ala Tyr Tyr Leu Ser Ile
130 135 140
Pro Pro Asp Ser Phe Ala Ala Val Cys His Gln Leu Glu Arg Ser Gly
145 150 155 160
Met Ala Glu Ser Thr Glu Glu Ala Trp Arg Arg Val Ile Ile Glu Lys
165 170 175
Pro Phe Gly His Asn Leu Glu Ser Ala His Glu Leu Asn Gln Leu Val
180 185 190
Asn Ala Val Phe Pro Glu Ser Ser Val Phe Arg Ile Asp His Tyr Leu
195 200 205
Gly Lys Glu Thr Val Gln Asn Ile Leu Ala Leu Arg Phe Ala Asn Gln
210 215 220
Leu Phe Glu Pro Leu Trp Asn Ser Asn Tyr Val Asp His Val Gln Ile
225 230 235 240
Thr Met Thr Glu Asp Ile Gly Leu Gly Gly Arg Ala Gly Tyr Tyr Asp
245 250 255
Gly Ile Gly Ala Ala Arg Asp Val Ile Gln Asn His Leu Ile Gln Leu
260 265 270
Leu Ala Leu Val Ala Met Glu Glu Pro Ile Ser Phe Val Pro Ala Gln
275 280 285
Leu Gln Ala Glu Lys Ile Lys Val Leu Ser Ala Thr Lys Pro Cys Tyr
290 295 300
Pro Leu Asp Lys Thr Ser Ala Arg Gly Gln Tyr Ala Ala Gly Trp Gln
305 310 315 320
Gly Ser Glu Leu Val Lys Gly Leu Arg Glu Glu Asp Gly Phe Asn Pro
325 330 335
Glu Ser Thr Thr Glu Thr Phe Ala Ala Cys Thr Leu Glu Ile Thr Ser
340 345 350
Arg Arg Trp Ala Gly Val Pro Phe Tyr Leu Arg Thr Gly Lys Arg Leu
355 360 365
Gly Arg Arg Val Thr Glu Ile Ala Val Val Phe Lys Asp Ala Pro His
370 375 380
Gln Pro Phe Asp Gly Asp Met Thr Val Ser Leu Gly Gln Asn Ala Ile
385 390 395 400
Val Ile Arg Val Gln Pro Asp Glu Gly Val Leu Ile Arg Phe Gly Ser
405 410 415
Lys Val Pro Gly Ser Ala Met Glu Val Arg Asp Val Asn Met Asp Phe
420 425 430
Ser Tyr Ser Glu Ser Phe Thr Glu Glu Ser Pro Glu Ala Tyr Glu Arg
435 440 445
Leu Ile Leu Asp Ala Leu Leu Asp Glu Ser Ser Leu Phe Pro Thr Asn
450 455 460
Glu Glu Val Glu Leu Ser Trp Lys Ile Leu Asp Pro Ile Leu Glu Ala
465 470 475 480
Trp Asp Ala Asp Gly Glu Pro Glu Asp Tyr Pro Ala Gly Thr Trp Gly
485 490 495
Pro Lys Ser Ala Asp Glu Met Leu Ser Arg Asn Gly His Thr Trp Arg
500 505 510
Arg Pro
<210> 27
<211> 680
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）（菌株ATCC 204508 / S288c）（面包酵母）
<400> 27
Met Thr Gln Phe Thr Asp Ile Asp Lys Leu Ala Val Ser Thr Ile Arg
1 5 10 15
Ile Leu Ala Val Asp Thr Val Ser Lys Ala Asn Ser Gly His Pro Gly
20 25 30
Ala Pro Leu Gly Met Ala Pro Ala Ala His Val Leu Trp Ser Gln Met
35 40 45
Arg Met Asn Pro Thr Asn Pro Asp Trp Ile Asn Arg Asp Arg Phe Val
50 55 60
Leu Ser Asn Gly His Ala Val Ala Leu Leu Tyr Ser Met Leu His Leu
65 70 75 80
Thr Gly Tyr Asp Leu Ser Ile Glu Asp Leu Lys Gln Phe Arg Gln Leu
85 90 95
Gly Ser Arg Thr Pro Gly His Pro Glu Phe Glu Leu Pro Gly Val Glu
100 105 110
Val Thr Thr Gly Pro Leu Gly Gln Gly Ile Ser Asn Ala Val Gly Met
115 120 125
Ala Met Ala Gln Ala Asn Leu Ala Ala Thr Tyr Asn Lys Pro Gly Phe
130 135 140
Thr Leu Ser Asp Asn Tyr Thr Tyr Val Phe Leu Gly Asp Gly Cys Leu
145 150 155 160
Gln Glu Gly Ile Ser Ser Glu Ala Ser Ser Leu Ala Gly His Leu Lys
165 170 175
Leu Gly Asn Leu Ile Ala Ile Tyr Asp Asp Asn Lys Ile Thr Ile Asp
180 185 190
Gly Ala Thr Ser Ile Ser Phe Asp Glu Asp Val Ala Lys Arg Tyr Glu
195 200 205
Ala Tyr Gly Trp Glu Val Leu Tyr Val Glu Asn Gly Asn Glu Asp Leu
210 215 220
Ala Gly Ile Ala Lys Ala Ile Ala Gln Ala Lys Leu Ser Lys Asp Lys
225 230 235 240
Pro Thr Leu Ile Lys Met Thr Thr Thr Ile Gly Tyr Gly Ser Leu His
245 250 255
Ala Gly Ser His Ser Val His Gly Ala Pro Leu Lys Ala Asp Asp Val
260 265 270
Lys Gln Leu Lys Ser Lys Phe Gly Phe Asn Pro Asp Lys Ser Phe Val
275 280 285
Val Pro Gln Glu Val Tyr Asp His Tyr Gln Lys Thr Ile Leu Lys Pro
290 295 300
Gly Val Glu Ala Asn Asn Lys Trp Asn Lys Leu Phe Ser Glu Tyr Gln
305 310 315 320
Lys Lys Phe Pro Glu Leu Gly Ala Glu Leu Ala Arg Arg Leu Ser Gly
325 330 335
Gln Leu Pro Ala Asn Trp Glu Ser Lys Leu Pro Thr Tyr Thr Ala Lys
340 345 350
Asp Ser Ala Val Ala Thr Arg Lys Leu Ser Glu Thr Val Leu Glu Asp
355 360 365
Val Tyr Asn Gln Leu Pro Glu Leu Ile Gly Gly Ser Ala Asp Leu Thr
370 375 380
Pro Ser Asn Leu Thr Arg Trp Lys Glu Ala Leu Asp Phe Gln Pro Pro
385 390 395 400
Ser Ser Gly Ser Gly Asn Tyr Ser Gly Arg Tyr Ile Arg Tyr Gly Ile
405 410 415
Arg Glu His Ala Met Gly Ala Ile Met Asn Gly Ile Ser Ala Phe Gly
420 425 430
Ala Asn Tyr Lys Pro Tyr Gly Gly Thr Phe Leu Asn Phe Val Ser Tyr
435 440 445
Ala Ala Gly Ala Val Arg Leu Ser Ala Leu Ser Gly His Pro Val Ile
450 455 460
Trp Val Ala Thr His Asp Ser Ile Gly Val Gly Glu Asp Gly Pro Thr
465 470 475 480
His Gln Pro Ile Glu Thr Leu Ala His Phe Arg Ser Leu Pro Asn Ile
485 490 495
Gln Val Trp Arg Pro Ala Asp Gly Asn Glu Val Ser Ala Ala Tyr Lys
500 505 510
Asn Ser Leu Glu Ser Lys His Thr Pro Ser Ile Ile Ala Leu Ser Arg
515 520 525
Gln Asn Leu Pro Gln Leu Glu Gly Ser Ser Ile Glu Ser Ala Ser Lys
530 535 540
Gly Gly Tyr Val Leu Gln Asp Val Ala Asn Pro Asp Ile Ile Leu Val
545 550 555 560
Ala Thr Gly Ser Glu Val Ser Leu Ser Val Glu Ala Ala Lys Thr Leu
565 570 575
Ala Ala Lys Asn Ile Lys Ala Arg Val Val Ser Leu Pro Asp Phe Phe
580 585 590
Thr Phe Asp Lys Gln Pro Leu Glu Tyr Arg Leu Ser Val Leu Pro Asp
595 600 605
Asn Val Pro Ile Met Ser Val Glu Val Leu Ala Thr Thr Cys Trp Gly
610 615 620
Lys Tyr Ala His Gln Ser Phe Gly Ile Asp Arg Phe Gly Ala Ser Gly
625 630 635 640
Lys Ala Pro Glu Val Phe Lys Phe Phe Gly Phe Thr Pro Glu Gly Val
645 650 655
Ala Glu Arg Ala Gln Lys Thr Ile Ala Phe Tyr Lys Gly Asp Lys Leu
660 665 670
Ile Ser Pro Leu Lys Lys Ala Phe
675 680
<210> 28
<211> 496
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）（菌株ATCC 204508 / S288c）（面包酵母）
<400> 28
Met Ser Met Ser Asn Ile Val Val Phe Gly Gly Asp Ser His Pro Glu
1 5 10 15
Leu Val Thr Lys Ile Cys Glu Asn Leu Asp Ile His Pro Ser Lys Val
20 25 30
Glu Leu Gly Lys Phe Ser Asn Gly Glu Thr Asn Ile Ala Leu Arg Glu
35 40 45
Ser Val Arg Glu Lys Asp Val Tyr Ile Ile Gln Ser Gly Cys Gly Gln
50 55 60
Val Asn Asp Thr Phe Met Gln Leu Leu Ile Leu Ile Ser Ala Cys Lys
65 70 75 80
Ser Ala Ser Ala Ser Arg Val Thr Ala Val Met Pro Tyr Leu Cys Tyr
85 90 95
Ser Arg Gln Pro Asp Ile Pro Tyr Thr Ala Lys Gly Ala Pro Ile Ile
100 105 110
Ser Lys Pro Lys Glu Asn Tyr Thr Phe Glu Ser His Pro Gly Thr Pro
115 120 125
Val Ser Ser Ser Leu Met Thr Gln Arg Pro Gly Ala Glu Ser Ser Leu
130 135 140
Lys Ser Leu Asp Ser Ala Ile Arg Ser Thr Ile Asn Leu Glu Asn Pro
145 150 155 160
Gln Pro Ile Arg Thr Pro Asn Ser Ser Ala Thr Ala Asn Asn Asn Phe
165 170 175
Asp Ile Lys Lys Thr Leu Ser Phe Ser Arg Ile Pro Met Ile Pro Gly
180 185 190
Gly Lys Leu Gln Asn Thr Ser Asn Ser Thr Asp Ala Gly Glu Leu Phe
195 200 205
Asn Ala Gln Asn Ala Gly Tyr Lys Leu Trp Val Val Gln Ala Gly Thr
210 215 220
Leu Ile Ala His Leu Leu Ser Ala Ala Gly Ala Asp His Val Ile Thr
225 230 235 240
Met Asp Leu His Asp Pro Gln Phe Pro Gly Phe Phe Asp Ile Pro Val
245 250 255
Asp Asn Leu Tyr Cys Lys Pro Ile Ala Gln Asn Tyr Ile Gln His Arg
260 265 270
Ile Pro Asp Tyr Gln Asp Ala Val Ile Val Ser Pro Asp Ala Gly Gly
275 280 285
Ala Lys Arg Ala Thr Ala Ile Ala Asp Ala Leu Glu Leu Ser Phe Ala
290 295 300
Leu Ile His Lys Glu Arg Arg Ser Gln Leu Leu Lys Gly Pro Pro Asp
305 310 315 320
Ala Thr Leu Thr Ser Gly Gly Ser Leu Pro Val Ser Pro Arg Pro Leu
325 330 335
Val Thr Thr Leu Val Ser Ser Gln Asn Thr Thr Ser Ser Gly Ala Thr
340 345 350
Gly Val Ala Ala Leu Glu Met Lys Lys Thr Thr Ser Thr Ser Ser Thr
355 360 365
Ser Ser Gln Ser Ser Asn Ser Ser Lys Phe Val Gln Thr Thr Met Leu
370 375 380
Val Gly Asp Val Arg Asn Lys Val Cys Ile Ile Val Asp Asp Leu Val
385 390 395 400
Asp Thr Ser Tyr Thr Ile Thr Arg Ala Ala Lys Leu Leu Lys Asp Gln
405 410 415
Gly Ser Thr Lys Val Tyr Ala Leu Ile Thr His Gly Val Phe Ser Gly
420 425 430
Asp Ala Leu Glu Arg Ile Gly Gln Ser Ser Ile Asp Lys Leu Ile Ile
435 440 445
Ser Asn Thr Val Pro Gln Asp Arg Thr Leu Gln Tyr Leu Gly Lys Asp
450 455 460
Arg Val Asp Val Ile Asp Val Ser Cys Ile Ile Gly Glu Ala Ile Arg
465 470 475 480
Arg Ile His Asn Gly Glu Ser Ile Ser Met Leu Phe Glu His Gly Trp
485 490 495
<210> 29
<211> 2397
<212> DNA
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 29
atggttttgc cgattctacc gttaattgat gatctggcct catggaatag taagaaggaa 60
tacgtttcac ttgttggtca ggtacttttg gatggctcga gcctgagtaa tgaagagatt 120
ctccagttct ccaaagagga agaagttcca ttggtggctt tgtccttgcc aagtggtaaa 180
ttcagcgatg atgaaatcat tgccttcttg aacaacggag tttcttctct gttcattgct 240
agccaagatg ctaaaacagc cgaacacttg gttgaacaat tgaatgtacc aaaggagcgt 300
gttgttgtgg aagagaacgg tgttttctcc aatcaattca tggtaaaaca aaaattctcg 360
caagataaaa ttgtgtccat aaagaaatta agcaaggata tgttgaccaa agaagtgctt 420
ggtgaagtac gtacagaccg tcctgacggt ttatatacca ccctagttgt cgaccaatat 480
gagcgttgtc tagggttggt gtattcttcg aagaaatcta tagcaaaggc catcgatttg 540
ggtcgtggcg tttattattc tcgttctagg aatgaaatct ggatcaaggg tgaaacttct 600
ggcaatggcc aaaagctttt acaaatctct actgactgtg attcggatgc cttaaagttt 660
atcgttgaac aagaaaacgt tggattttgc cacttggaga ccatgtcttg ctttggtgaa 720
ttcaagcatg gtttggtggg gctagaatct ttactaaaac aaaggctaca ggacgctcca 780
gaggaatctt atactagaag actattcaac gactctgcat tgttagatgc caagatcaag 840
gaagaagctg aagaactgac tgaggcaaag ggtaagaagg agctttcttg ggaggctgcc 900
gatttgttct actttgcact ggccaaatta gtggccaacg atgtttcatt gaaggacgtc 960
gagaataatc tgaatatgaa gcatctgaag gttacaagac ggaaaggtga tgctaagcca 1020
aagtttgttg gacaaccaaa ggctgaagaa gaaaaactga ccggtccaat tcacttggac 1080
gtggtgaagg cttccgacaa agttggtgtg cagaaggctt tgagcagacc aatccaaaag 1140
acttctgaaa ttatgcattt agtcaatccg atcatcgaaa atgttagaga caaaggtaac 1200
tctgcccttt tggagtacac agaaaagttt gatggtgtaa aattatccaa tcctgttctt 1260
aatgctccat tcccagaaga atactttgaa ggtttaaccg aggaaatgaa ggaagctttg 1320
gacctttcaa ttgaaaacgt ccgcaaattc catgctgctc aattgccaac agagactctt 1380
gaagttgaaa cccaacctgg tgtcttgtgt tccagattcc ctcgtcctat tgaaaaagtt 1440
ggtttgtata tccctggtgg cactgccatt ttaccaagta ctgcattaat gcttggtgtt 1500
ccagcacaag ttgcccaatg taaggagatt gtgtttgcat ctccaccaag aaaatctgat 1560
ggtaaagttt cacccgaagt tgtttatgtc gcagaaaaag ttggcgcttc caagattgtt 1620
ctagctggtg gtgcccaagc cgttgctgct atggcttacg ggacagaaac tattcctaaa 1680
gtggataaga tcttgggtcc aggtaatcaa tttgtgactg ccgccaaaat gtatgttcaa 1740
aatgacactc aagctctatg ttccattgat atgccagctg gcccaagtga agttttggtt 1800
attgccgatg aagatgccga tgtggatttt gttgcaagtg atttgctatc gcaagctgaa 1860
cacggtattg actcccaagt tatccttgtt ggtgttaact tgagcgaaaa gaaaattcaa 1920
gagattcaag atgctgtcca caatcaagct ttacaactgc cacgtgtgga tattgttcgt 1980
aaatgtattg ctcacagtac gatcgttctt tgtgacggtt acgaagaagc ccttgaaatg 2040
tccaaccaat atgcaccaga acatttgatt ctacaaatcg ccaatgctaa cgattatgtt 2100
aaattggttg acaatgcagg gtccgtattt gtgggtgctt acactccaga atcgtgcggt 2160
gactattcaa gtggtactaa ccatacatta ccaacctatg gttacgctag gcagtacagt 2220
ggtgccaaca ctgcaacctt ccaaaagttt atcactgccc aaaacattac ccctgaaggt 2280
ttagaaaaca tcggtagagc tgttatgtgc gttgccaaga aggagggtct agacggtcac 2340
agaaacgctg tgaaaatcag aatgagtaag cttgggttga tcccaaagga tttccag 2397
<210> 30
<211> 843
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 30
atgttgaaaa tcgctgtccc aaacaagggc tcgctgtccg agcgcgccat ggaaatcctc 60
gccgaagcag gctacgcagg ccgtggagat tccaaatccc tcaacgtttt tgatgaagca 120
aacaacgttg aattcttctt ccttcgccct aaagatatcg ccatctacgt tgcaggtggc 180
cagctcgatt tgggtatcac cggccgcgac cttgctcgcg attcccaggc tgatgtccac 240
gaagttcttt ccctcggctt cggttcctcc accttccgtt acgcagcacc agctgatgaa 300
gagtggagca tcgaaaagct cgacggcaag cgcatcgcta cctcttaccc caaccttgtt 360
cgcgatgacc tcgcagcacg tgggctttcc gctgaggtgc tccgcctcga cggtgcagta 420
gaggtatcca tcaagcttgg tgtcgcagat gccatcgccg atgttgtatc caccggccgc 480
acgctgcgtc agcaaggtct tgcacctttc ggcgaggttc tgtgcacctc tgaggctgtc 540
attgttggcc gcaaggatga aaaggtcacc ccagagcagc agatcctgct tcgccgcatc 600
cagggaattt tgcacgcgca gaacttcctc atgctggatt acaacgtcga ccgcgacaac 660
ctggacgctg ccactgcagt aaccccaggc ctatccggcc caacggtatc cccactggca 720
cgcgacaact gggttgctgt acgcgccatg gtgccacgca ggtcagctaa cgccatcatg 780
gataagcttg ctggactcgg cgctgaagcc atcctggctt ctgaaatccg catcgcccgc 840
atc 843
<210> 31
<211> 261
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）（菌株ATCC 204508 / S288c）（面包酵母）
<400> 31
Met Thr Lys Phe Ile Gly Cys Ile Asp Leu His Asn Gly Glu Val Lys
1 5 10 15
Gln Ile Val Gly Gly Thr Leu Thr Ser Lys Lys Glu Asp Val Pro Lys
20 25 30
Thr Asn Phe Val Ser Gln His Pro Ser Ser Tyr Tyr Ala Lys Leu Tyr
35 40 45
Lys Asp Arg Asp Val Gln Gly Cys His Val Ile Lys Leu Gly Pro Asn
50 55 60
Asn Asp Asp Ala Ala Arg Glu Ala Leu Gln Glu Ser Pro Gln Phe Leu
65 70 75 80
Gln Val Gly Gly Gly Ile Asn Asp Thr Asn Cys Leu Glu Trp Leu Lys
85 90 95
Trp Ala Ser Lys Val Ile Val Thr Ser Trp Leu Phe Thr Lys Glu Gly
100 105 110
His Phe Gln Leu Lys Arg Leu Glu Arg Leu Thr Glu Leu Cys Gly Lys
115 120 125
Asp Arg Ile Val Val Asp Leu Ser Cys Arg Lys Thr Gln Asp Gly Arg
130 135 140
Trp Ile Val Ala Met Asn Lys Trp Gln Thr Leu Thr Asp Leu Glu Leu
145 150 155 160
Asn Ala Asp Thr Phe Arg Glu Leu Arg Lys Tyr Thr Asn Glu Phe Leu
165 170 175
Ile His Ala Ala Asp Val Glu Gly Leu Cys Gly Gly Ile Asp Glu Leu
180 185 190
Leu Val Ser Lys Leu Phe Glu Trp Thr Lys Asp Tyr Asp Asp Leu Lys
195 200 205
Ile Val Tyr Ala Gly Gly Ala Lys Ser Val Asp Asp Leu Lys Leu Val
210 215 220
Asp Glu Leu Ser His Gly Lys Val Asp Leu Thr Phe Gly Ser Ser Leu
225 230 235 240
Asp Ile Phe Gly Gly Asn Leu Val Lys Phe Glu Asp Cys Cys Arg Trp
245 250 255
Asn Glu Lys Gln Gly
260
<210> 32
<211> 248
<212> PRT
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 32
Met Ala Arg Thr Phe Phe Val Gly Gly Asn Phe Lys Leu Asn Gly Ser
1 5 10 15
Lys Gln Ser Ile Lys Glu Ile Val Glu Arg Leu Asn Thr Ala Ser Ile
20 25 30
Pro Glu Asn Val Glu Val Val Ile Cys Pro Pro Ala Thr Tyr Leu Asp
35 40 45
Tyr Ser Val Ser Leu Val Lys Lys Pro Gln Val Thr Val Gly Ala Gln
50 55 60
Asn Ala Tyr Leu Lys Ala Ser Gly Ala Phe Thr Gly Glu Asn Ser Val
65 70 75 80
Asp Gln Ile Lys Asp Val Gly Ala Lys Trp Val Ile Leu Gly His Ser
85 90 95
Glu Arg Arg Ser Tyr Phe His Glu Asp Asp Lys Phe Ile Ala Asp Lys
100 105 110
Thr Lys Phe Ala Leu Gly Gln Gly Val Gly Val Ile Leu Cys Ile Gly
115 120 125
Glu Thr Leu Glu Glu Lys Lys Ala Gly Lys Thr Leu Asp Val Val Glu
130 135 140
Arg Gln Leu Asn Ala Val Leu Glu Glu Val Lys Asp Trp Thr Asn Val
145 150 155 160
Val Val Ala Tyr Glu Pro Val Trp Ala Val Gly Thr Gly Leu Ala Ala
165 170 175
Thr Pro Glu Asp Ala Gln Asp Ile His Ala Ser Ile Arg Lys Phe Leu
180 185 190
Ala Ser Lys Leu Gly Asp Lys Ala Ala Ser Glu Leu Arg Ile Leu Tyr
195 200 205
Gly Gly Ser Ala Asn Gly Ser Asn Ala Val Thr Phe Lys Asp Lys Ala
210 215 220
Asp Val Asp Gly Phe Leu Val Gly Gly Ala Ser Leu Lys Pro Glu Phe
225 230 235 240
Val Asp Ile Ile Asn Ser Arg Asn
245
<210> 33
<211> 408
<212> PRT
<213> 根际假单胞菌（Pseudomonas rhizosphaerae）
<400> 33
Met Thr Leu Ser Thr Ala Asp Thr His Arg Leu Asp Asp Phe Trp Gln
1 5 10 15
His Cys Leu Lys His Gln Phe Phe Asn Ile Gly Tyr Pro Glu Asn Ala
20 25 30
Asp Phe Asp Tyr Ser Ala Leu Glu Arg Phe Leu Arg Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Ser Glu His Ser Asn Tyr Val Leu Asn Ser Phe
50 55 60
Asp Phe Glu Lys Glu Val Met Ala Tyr Phe Ala Asp Leu Phe Glu Ile
65 70 75 80
Pro Arg Glu Asp Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Ala Arg Glu Leu Phe Pro Asp Gly Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Thr His Tyr Ser Val Ala Lys Ile Val Lys
115 120 125
Leu Leu Arg Ile Lys Cys Arg Ala Val Asn Ala Leu Pro Thr Gly Glu
130 135 140
Ile Asp Tyr Asp Asp Leu Leu Ala Lys Ile Ala Ala Asp Gly Glu Arg
145 150 155 160
His Pro Ile Ile Phe Ala Asn Ile Gly Thr Thr Met Arg Gly Ala Val
165 170 175
Asp Asp Ile Ala Val Ile Gln Gln Arg Leu Gln Asp Ala Gly Ile Ala
180 185 190
Arg Arg Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asp Ala Pro Gln Pro Phe Thr Phe Ala Asp Gly Ile
210 215 220
Asp Ser Ile Cys Val Ser Gly His Lys Met Ile Gly Ser Pro Ile Pro
225 230 235 240
Cys Gly Ile Val Val Ala Lys Arg Arg Asn Val Ala Arg Ile Ser Val
245 250 255
Glu Val Asp Tyr Ile Ser Ala Ser Asp Lys Thr Ile Ser Gly Ser Arg
260 265 270
Asn Gly His Thr Pro Met Ile Met Trp Ala Ala Leu Arg Ser His Ser
275 280 285
Ser Ala Gln Trp Arg Arg Arg Val Glu Arg Ser Leu Ala Ala Ala Gln
290 295 300
Tyr Ala Val Asn Arg Leu Gln Ala Gly Gly Val Lys Ala Trp Arg Asn
305 310 315 320
Pro Asn Ser Ile Thr Val Val Phe Pro Cys Pro Ser Ala Asp Ile Ala
325 330 335
Arg Lys Tyr Gly Leu Ala Thr Ser Gly Asp Thr Ala His Leu Ile Thr
340 345 350
Thr Pro His His Arg Asp Asn Arg Ala Ile Asp Ala Leu Ile Asp Glu
355 360 365
Val Ile Ala Asp Ala Arg Pro Glu Val Trp Arg Ala Thr Leu Gln Arg
370 375 380
Ser Trp Gln Gly Ser Ala Gln Arg Leu Pro Arg Thr Ala Ser Trp Asn
385 390 395 400
Gln Ala Ala Gly Leu Gly Arg Phe
405
<210> 34
<211> 427
<212> PRT
<213> 恶臭假单胞菌（Pseudomonas putida）（铁荚膜节杆菌（Arthrobactersiderocapsulatus））
<400> 34
Met Thr Phe Ser Pro Ala Asp His Lys Arg Leu Glu Ala Phe Trp Gln
1 5 10 15
Tyr Cys Leu Thr His Gln Tyr Phe Asn Val Gly Tyr Pro Glu Ser Ala
20 25 30
Asp Phe Asp Tyr Ser Leu Leu His Arg Phe Met Arg Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Asn Glu Pro Ser Asn Tyr Leu Leu Asn Ser Phe
50 55 60
Asp Phe Glu Arg Glu Val Met Arg Phe Phe Ala Glu Leu Phe His Ile
65 70 75 80
Pro Phe Glu Asp Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Ala Arg Glu Leu Phe Pro Asp Ala Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Thr His Tyr Ser Val Ala Lys Ile Val Arg
115 120 125
Leu Leu Arg Ile Lys Ala Gln Val Val Glu Ser Gln Ala Asn Gly Glu
130 135 140
Met Asp Tyr Asp Asp Leu Val Ala Arg Ile Ala Ala Asp Gly Glu Arg
145 150 155 160
His Pro Ile Ile Phe Ala Asn Ile Gly Thr Thr Leu Arg Gly Ala Thr
165 170 175
Asp Asn Ile Ala Val Ile Gln Gln Arg Leu Ala Gln Ala Gly Ile Arg
180 185 190
Arg Glu Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asp Ala Pro Glu Pro Tyr Ser Phe Ala Asp Gly Ile
210 215 220
Asp Ser Ile Cys Val Ser Gly His Lys Met Ile Gly Ser Pro Met Pro
225 230 235 240
Cys Gly Ile Val Val Ala Arg Arg His Asn Val Glu Arg Ile Ser Val
245 250 255
Glu Ile Asp Tyr Ile Ser Ala Arg Asp Gln Thr Ile Ser Gly Ser Arg
260 265 270
Asn Gly His Thr Pro Leu Met Met Trp Ala Ala Leu Cys Ser Arg Ser
275 280 285
Arg Glu Asp Trp Arg Ala Arg Ile Gln Arg Cys Leu Asp Leu Ala Gln
290 295 300
His Ala Val Asp Arg Leu Arg Ala Ala Gly Ile Glu Ala Trp Arg Asn
305 310 315 320
Pro Asn Ser Ile Thr Val Val Phe Pro Cys Pro Ser Ala Ser Val Trp
325 330 335
Lys Arg His Cys Leu Ala Thr Ser Gly Asp Thr Ala His Leu Ile Thr
340 345 350
Thr Ala His His Gln Asp Ser Thr Gln Ile Asp Ala Leu Leu Asp Glu
355 360 365
Leu Ile Ala Asp Leu Lys Ala Arg Thr Arg Tyr Pro Leu Gln Ala Pro
370 375 380
Leu Val Glu Gly Leu Gly Ser Arg Thr Arg Val Tyr Ser Thr Ala Gly
385 390 395 400
Gln Pro Arg Pro Glu Pro Leu Pro Asp His Ala Arg Pro Ala Arg Asp
405 410 415
Ser Gly Leu Ala Arg Leu Lys Asp Ser Glu Ile
420 425
<210> 35
<211> 378
<212> PRT
<213> 巴氏柠檬酸杆菌（Citrobacter pasteurii）
<400> 35
Met Thr Leu Ser Ile Val Asp Gln Asn Lys Leu Asp Ala Phe Trp Ser
1 5 10 15
Tyr Cys Val Lys Asn Gln Tyr Phe Asn Ile Gly Tyr Pro Glu Ser Ala
20 25 30
Asp Phe Asp Tyr Thr Ile Leu Glu Arg Phe Met Arg Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Gly Glu Tyr Cys Asn Tyr Leu Leu Asn Ser Phe
50 55 60
Asp Phe Glu Lys Glu Val Met Glu Tyr Phe Ala Arg Ile Phe Lys Ile
65 70 75 80
Pro Phe Glu Glu Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Gly Arg Glu Leu Phe Pro Glu Gly Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Thr His Tyr Ser Val Ala Lys Ile Val Lys
115 120 125
Leu Leu Arg Ile Lys Ser Ser Leu Val Glu Ser Gln Pro Asn Gly Glu
130 135 140
Met Asp Tyr Asp Asp Leu Ile Arg Lys Ile Gln Arg Asp Asn Glu Glu
145 150 155 160
His Pro Ile Ile Phe Ala Asn Ile Gly Thr Thr Val Arg Gly Ala Ile
165 170 175
Asp Asn Ile Ala Glu Ile Gln Gln Arg Ile Gly Gln Leu Gly Ile Lys
180 185 190
Arg Asp Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asn Asp Pro Gln Pro Phe Asn Phe Ala Asp Gly Ile
210 215 220
Asp Ser Ile Gly Val Ser Gly His Lys Met Ile Gly Ser Pro Ile Pro
225 230 235 240
Cys Gly Ile Val Val Ala Lys Arg Lys Asn Val Asp Arg Ile Ser Val
245 250 255
Glu Ile Asp Tyr Ile Ser Ala His Asp Lys Thr Ile Ser Gly Ser Arg
260 265 270
Asn Gly His Thr Pro Leu Met Met Trp Glu Ala Ile Arg Ser His Ser
275 280 285
Trp Ser Asp Trp Gln Arg Arg Ile Glu His Ser Leu Asn Met Ala Gln
290 295 300
Tyr Ala Val Asp Arg Leu Gln Ala Ala Gly Ile Asp Ala Trp Arg Asn
305 310 315 320
Lys Asn Ser Ile Thr Val Val Phe Pro Cys Pro Ser Glu Ala Val Trp
325 330 335
Lys Lys His Cys Leu Ala Thr Ser Gly Asp Ile Ala His Leu Ile Thr
340 345 350
Thr Ala His His Leu Asp Ser Ser Lys Ile Asp Glu Leu Ile Asp Asp
355 360 365
Val Ile Ala Asp Leu Asn Gln Gln Ala Ala
370 375
<210> 36
<211> 387
<212> PRT
<213> 溶血色杆菌（Chromobacterium haemolyticum）
<400> 36
Met Ser Leu Ser Ser Leu Asp Gln Asn Arg Ile Glu Ser Phe Trp Gln
1 5 10 15
Tyr Cys Leu Gln His Gln Tyr Phe Asn Leu Ala Tyr Pro Glu Ser Ala
20 25 30
Asp Phe Asp Tyr Thr Pro Leu His Arg Phe Leu Arg Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Asn Glu Ser Ser Asn Tyr Leu Leu Asn Ser Phe
50 55 60
Asp Phe Glu Arg Glu Val Met His Phe Phe Ala Glu Leu Phe His Ile
65 70 75 80
Pro Phe Asp Glu Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Ala Arg Glu Leu Phe Pro Asp Ala Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Ser His Tyr Ser Val Ala Lys Ile Ile Lys
115 120 125
Leu Leu Arg Ile Lys Ser Arg Ala Val Asp Ser Leu Pro Ser Gly Glu
130 135 140
Ile Asp Tyr Asp Asp Leu Val Ala Lys Ile Gln Gln Asp Gln Glu Arg
145 150 155 160
His Pro Ile Val Phe Val Asn Val Gly Thr Thr Met Lys Gly Ala Val
165 170 175
Asp Asp Ile Gly Ile Ile Gln Asp Lys Leu Ala Gln Ala Gly Ile Pro
180 185 190
Arg Arg Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asp Ala Pro Gln Pro Tyr Ser Phe Ala Asp Gly Ile
210 215 220
Asp Ser Ile Ser Val Ser Gly His Lys Met Ile Gly Ser Pro Met Pro
225 230 235 240
Cys Gly Ile Val Leu Ala Lys Arg Ser Asn Val Ser Arg Ile Ser Val
245 250 255
Glu Ile Asp Tyr Ile Ser Ala Arg Asp Gln Thr Ile Ser Gly Ser Arg
260 265 270
Asn Gly His Thr Pro Met Met Leu Trp Ala Ala Ile Lys Ser Arg Pro
275 280 285
Leu Ala Glu Trp Arg Arg Lys Val Arg His Cys Leu Asp Met Ala Gln
290 295 300
Tyr Ala Ile Asp Arg Phe Arg Ala Ala Gly Ile Gln Ala Trp Arg Cys
305 310 315 320
Gln Asn Ser Ile Thr Val Val Phe Pro Ser Pro Ser Glu Pro Val Cys
325 330 335
Asp Lys His Gly Leu Ala Arg Ser Gly Gly Ala Ala His Leu Ile Thr
340 345 350
Thr Pro His His His Asp Ser Gln Arg Leu Asp Arg Leu Ile Asp Asp
355 360 365
Ile Ile Gln Asp Leu Gly Ala Val Thr Ala Pro Ala Gly Ala Ala Met
370 375 380
Ser Ala Ala
385
<210> 37
<211> 311
<212> PRT
<213> 罗伊乳杆菌（Lactobacillus reuteri）
<400> 37
Met Ser Glu Leu Asp Thr Lys Leu His Lys Leu Gly Val Asp Arg Ile
1 5 10 15
Ala Ile Ser Pro Tyr Lys Gln Trp Ser Arg Gly Tyr Met Glu Pro Gly
20 25 30
Asn Ile Gly Asn Gly Tyr Val Thr Gly Leu Lys Val Asp Ala Gly Val
35 40 45
Arg Asp Lys Thr Asp Asp Glu Val Leu Asp Gly Ile Val Ser Tyr Asp
50 55 60
Arg Ala Glu Thr Lys Asn Ala Tyr Ile Gly Gln Ile Asn Met Thr Thr
65 70 75 80
Ala Ser Ser Phe Thr Gly Pro Gln Gly His Cys Ile Gly Tyr Asp Leu
85 90 95
Leu Arg Asn Pro Glu Val Asp Thr Ala Glu Pro Leu Phe Thr Val Lys
100 105 110
Gln Trp Asp Gly Ser Glu Leu Pro Ile Tyr Asp Ala Lys Pro Leu Gln
115 120 125
Asp Ser Leu Val Glu Tyr Phe Gly Thr Asn Asn Asn Arg Arg His Tyr
130 135 140
Pro Ala Pro Gly Ser Phe Ile Val Cys Ala Asn Lys Gly Val Thr Ala
145 150 155 160
Glu Arg Pro Met Asn Asp Ser Asp Met Lys Pro Gly Gln Gly Tyr Gly
165 170 175
Val Trp Ser Ala Ile Ala Leu Ser Phe Ala Lys Asp Pro Ala Lys Asp
180 185 190
Ser Ser Met Phe Ile Glu Asp Ala Gly Val Trp Glu Thr Pro Asn Glu
195 200 205
Asp Glu Leu Ile Glu Tyr Leu Lys Gly Arg Arg Lys Ala Ile Ala Lys
210 215 220
Ser Ile Ala Glu Cys Gly Gln Asp Ala Asn Thr Ser Phe Lys Gly Ser
225 230 235 240
Trp Ile Gly Phe Ala His Ala Met Met Glu Pro Gly Gln Ile Gly Asn
245 250 255
Ala Ile Thr Val Ala Pro Tyr Ile Ser Met Pro Val Asp Ser Ile Pro
260 265 270
Gly Gly Ser Ile Leu Thr Pro Asp Thr Asp Met Asp Ile Met Glu Asn
275 280 285
Leu Thr Met Pro Glu Trp Leu Asp Lys Met Glu Tyr Lys Ser Leu Thr
290 295 300
Ala Asn Gly Ala Ile Lys Tyr
305 310
<210> 38
<211> 390
<212> PRT
<213> 杀鲑气单胞菌溶果胶亚种（Aeromonas salmonicida subsp. pectinolytica）34mel
<400> 38
Met Cys Arg His Ala Cys Ile Ala Leu Ala Met Gly Met Ile Met Gly
1 5 10 15
Leu Ser Ser Glu Asp Ala Gly Lys Ile Glu Ser Phe Trp Arg Tyr Cys
20 25 30
Val Gln His Gln Tyr Phe Asn Ile Gly Tyr Pro Glu Ala Ala Asp Phe
35 40 45
Asp Tyr Ser Ala Leu Asn Arg Phe Leu Asn Phe Ser Ile Asn Asn Cys
50 55 60
Gly Asp Trp Ser Gln Gln Ser Asn Tyr Leu Leu Asn Ser Phe Asp Phe
65 70 75 80
Glu Arg Glu Val Met Gln Phe Phe Ala Thr Leu Phe Cys Ile Pro Phe
85 90 95
Glu Gln Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly Asn Met
100 105 110
Phe Gly Cys Tyr Leu Ala Arg Glu Leu Phe Pro Glu Ala Thr Leu Tyr
115 120 125
Tyr Ser Lys Asp Thr His Tyr Ser Val Ala Lys Ile Ile Arg Leu Leu
130 135 140
Arg Val Lys Ser Cys Met Val Asp Ser Leu Pro Asn Gly Glu Met Asn
145 150 155 160
Tyr Asp Asp Leu Ile Asn Arg Ile Arg Leu Asp Gly Glu Arg His Pro
165 170 175
Ile Ile Phe Ala Asn Ile Gly Thr Thr Met Thr Gly Ala Thr Asp Asn
180 185 190
Ile Ala Thr Ile Gln Arg Arg Leu Lys Lys Ile Gly Ile Thr Lys Gly
195 200 205
Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile Leu Pro
210 215 220
Phe Ile Asp Asn Pro Gln Pro Phe Ser Phe Ala Asp Gly Val Asp Ser
225 230 235 240
Ile Ser Val Ser Gly His Lys Met Ile Gly Ser Pro Ile Pro Cys Gly
245 250 255
Ile Val Leu Ala Arg Arg Lys His Val Glu His Val Ser Val Glu Ile
260 265 270
Asp Tyr Ile Ser Ala Cys Asp Gln Thr Ile Ser Gly Ser Arg Asn Gly
275 280 285
Tyr Thr Pro Leu Leu Leu Trp Met Ala Ile Lys Ser Arg Ser Phe Ser
290 295 300
Asp Trp Arg Gln Arg Thr Gln His Cys Leu Asp Met Ala Gln Tyr Val
305 310 315 320
Ile Glu Arg Phe His Ala Lys Gly Ile His Ala Trp Arg Asn Pro Asn
325 330 335
Ser Ile Thr Val Val Phe Pro Lys Pro Ala Asp His Ile Trp Lys Lys
340 345 350
His Cys Leu Ala Thr Ser Gly Lys Ile Ser His Ile Ile Thr Met Pro
355 360 365
His His Thr Gly Lys Glu Thr Leu Asp Arg Val Ile Asn Asp Ile Ala
370 375 380
Leu Asp Arg Glu Pro Lys
385 390
<210> 39
<211> 378
<212> PRT
<213> 耐冷摩根氏菌（Morganella psychrotolerans）
<400> 39
Met Thr Leu Ser Ile Asn Asp Gln Asn Lys Leu Asp Ala Phe Trp Ala
1 5 10 15
Tyr Cys Val Lys Asn Gln Tyr Phe Asn Ile Gly Tyr Pro Glu Ser Ala
20 25 30
Asp Phe Asp Tyr Thr Asn Leu Glu Arg Phe Leu Arg Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Gly Glu Tyr Cys Asn Tyr Leu Leu Asn Ser Phe
50 55 60
Asp Phe Glu Lys Glu Val Met Glu Tyr Phe Ala Asp Leu Phe Lys Ile
65 70 75 80
Pro Phe Glu Lys Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Gly Arg Glu Ile Phe Pro Asp Gly Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Thr His Tyr Ser Val Ala Lys Ile Val Lys
115 120 125
Leu Leu Arg Ile Lys Ser Gln Val Val Glu Ala Gln Pro Asn Gly Glu
130 135 140
Ile Asp Tyr Asp Asp Leu Met Lys Lys Ile Ala Ala Asp Lys Glu Ala
145 150 155 160
His Pro Ile Ile Phe Ala Asn Ile Gly Thr Thr Val Arg Gly Ala Ile
165 170 175
Asp Asp Ile Thr Glu Ile Gln Lys Arg Met Lys Ala Ala Gly Ile Lys
180 185 190
Arg Glu Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asp Glu Pro Gln Ala Phe Thr Phe Ala Asp Gly Ile
210 215 220
Asp Ser Ile Gly Val Ser Gly His Lys Met Ile Gly Ser Pro Ile Pro
225 230 235 240
Cys Gly Ile Val Val Ala Lys Lys Glu Asn Val Asp Arg Ile Ser Val
245 250 255
Glu Ile Asp Tyr Ile Ser Ala His Asp Lys Thr Ile Thr Gly Ser Arg
260 265 270
Asn Gly His Thr Pro Leu Met Leu Trp Glu Ala Val Arg Ala His Ser
275 280 285
Thr Glu Asp Trp Lys Arg Arg Ile Gly Arg Ser Leu Asp Met Ala Gln
290 295 300
Tyr Ala Val Asp Arg Leu Gln Lys Ala Gly Ile Asn Ala Trp Arg Asn
305 310 315 320
Lys Asn Ser Ile Thr Val Val Phe Pro Cys Pro Ser Glu Arg Val Trp
325 330 335
Lys Glu His Cys Leu Ala Thr Ser Gly Asn Asp Ala His Leu Ile Thr
340 345 350
Thr Ala His His Leu Asp Thr Ala Gln Ile Asp Ala Leu Ile Asp Asp
355 360 365
Val Ile Ala Asp Ala Lys Leu His Ala Ala
370 375
<210> 40
<211> 311
<212> PRT
<213> 食果糖乳杆菌（Lactobacillus fructivorans）
<400> 40
Met Ala Lys Ile Asp Lys Ile Leu Asn Gln Glu Gly Ile Asp Arg Ile
1 5 10 15
Ala Ile Asn Pro Tyr Gln Lys Tyr Ser Arg Gly Tyr Met Glu Pro Gly
20 25 30
Asn Leu Gly Gly Gly Tyr Val Thr Gly Leu Lys Val Asp Ala Gly Thr
35 40 45
Arg Glu Lys Thr Asp Asp Ser Met Leu Asp Gly Ile Val Ser Tyr Asp
50 55 60
Arg Ala Glu Cys Lys Asn Ala Tyr Ile Gly Gln Ile Asn Met Met Thr
65 70 75 80
Ala Ser Ser Phe Thr Gly Val Gln Gly His Ile Leu Gly Tyr Asp Leu
85 90 95
Leu Arg Asn Pro Ala Val Asp Lys Ala Gln Pro Leu Phe Tyr Glu Thr
100 105 110
Gln Trp Asp Gly Ser Lys Leu Pro Ile Tyr Asp Gly Lys Pro Leu Gln
115 120 125
Asp Ser Leu Val Glu Phe Phe Gly Thr Ala Asp Asn Arg Arg His Tyr
130 135 140
Pro Ala Pro Gly Ser Phe Ile Val Cys Ala Asn Lys Gly Val Thr Ala
145 150 155 160
Glu Arg Pro Leu Glu Asp Arg Pro Leu Asn Pro Gly Glu Ala Tyr Gly
165 170 175
Val Trp Ser Ala Ile Ala Ile Ser Ile Ala Lys Asp Pro Val His Asn
180 185 190
Ser Ser Met Phe Ile Glu Asp Ala Gly Thr Trp Asn Thr Pro Asn Glu
195 200 205
Asp Asp Leu Asn Glu Phe Leu Tyr His Arg Arg Glu Ala Ile Ala Arg
210 215 220
Ser Ile Ala Gln Cys Gly Gln Asp Ala Ser Thr Ser Phe Ala Ser Ser
225 230 235 240
Trp Ile Gly Phe Ala His Val Met Met Lys Pro Gly Glu Ile Gly Asn
245 250 255
Ala Ile Thr Val Gly Pro Tyr Phe Ser Met Pro Val Asp Ala Val Pro
260 265 270
Gly Gly Ser Ile Leu Thr Pro Asp Val Asp Met Asn Ile Met Glu Asp
275 280 285
Leu Ser Leu Pro Glu Trp Leu Glu Lys Met Gly Tyr Gln Ser Ile Val
290 295 300
Glu Asn Gln Asp Ile Gln Tyr
305 310
<210> 41
<211> 315
<212> PRT
<213> 大肠杆菌（Escherichia coli）（菌株K12）
<400> 41
Met Pro Asp Met Lys Leu Phe Ala Gly Asn Ala Thr Pro Glu Leu Ala
1 5 10 15
Gln Arg Ile Ala Asn Arg Leu Tyr Thr Ser Leu Gly Asp Ala Ala Val
20 25 30
Gly Arg Phe Ser Asp Gly Glu Val Ser Val Gln Ile Asn Glu Asn Val
35 40 45
Arg Gly Gly Asp Ile Phe Ile Ile Gln Ser Thr Cys Ala Pro Thr Asn
50 55 60
Asp Asn Leu Met Glu Leu Val Val Met Val Asp Ala Leu Arg Arg Ala
65 70 75 80
Ser Ala Gly Arg Ile Thr Ala Val Ile Pro Tyr Phe Gly Tyr Ala Arg
85 90 95
Gln Asp Arg Arg Val Arg Ser Ala Arg Val Pro Ile Thr Ala Lys Val
100 105 110
Val Ala Asp Phe Leu Ser Ser Val Gly Val Asp Arg Val Leu Thr Val
115 120 125
Asp Leu His Ala Glu Gln Ile Gln Gly Phe Phe Asp Val Pro Val Asp
130 135 140
Asn Val Phe Gly Ser Pro Ile Leu Leu Glu Asp Met Leu Gln Leu Asn
145 150 155 160
Leu Asp Asn Pro Ile Val Val Ser Pro Asp Ile Gly Gly Val Val Arg
165 170 175
Ala Arg Ala Ile Ala Lys Leu Leu Asn Asp Thr Asp Met Ala Ile Ile
180 185 190
Asp Lys Arg Arg Pro Arg Ala Asn Val Ser Gln Val Met His Ile Ile
195 200 205
Gly Asp Val Ala Gly Arg Asp Cys Val Leu Val Asp Asp Met Ile Asp
210 215 220
Thr Gly Gly Thr Leu Cys Lys Ala Ala Glu Ala Leu Lys Glu Arg Gly
225 230 235 240
Ala Lys Arg Val Phe Ala Tyr Ala Thr His Pro Ile Phe Ser Gly Asn
245 250 255
Ala Ala Asn Asn Leu Arg Asn Ser Val Ile Asp Glu Val Val Val Cys
260 265 270
Asp Thr Ile Pro Leu Ser Asp Glu Ile Lys Ser Leu Pro Asn Val Arg
275 280 285
Thr Leu Thr Leu Ser Gly Met Leu Ala Glu Ala Ile Arg Arg Ile Ser
290 295 300
Asn Glu Glu Ser Ile Ser Ala Met Phe Glu His
305 310 315
<210> 42
<211> 337
<212> PRT
<213> 拟南芥（Arabidopsis thaliana）（水芹鼠耳草（Mouse-ear cress））
<400> 42
Met Ser Glu Asn Ala Ala Asn Asn Ile Met Glu Thr Lys Ile Cys Thr
1 5 10 15
Asp Ala Ile Val Ser Glu Leu Gln Lys Lys Lys Val His Leu Phe Tyr
20 25 30
Cys Leu Glu Cys Glu Glu Leu Ala Arg Asn Ile Ala Ala Glu Ser Asp
35 40 45
His Ile Thr Leu Gln Ser Ile Asn Trp Arg Ser Phe Ala Asp Gly Phe
50 55 60
Pro Asn Leu Phe Ile Asn Asn Ala His Asp Ile Arg Gly Gln His Val
65 70 75 80
Ala Phe Leu Ala Ser Phe Ser Ser Pro Ala Val Ile Phe Glu Gln Ile
85 90 95
Ser Val Ile Tyr Leu Leu Pro Arg Leu Phe Val Ala Ser Phe Thr Leu
100 105 110
Val Leu Pro Phe Phe Pro Thr Gly Ser Phe Glu Arg Met Glu Glu Glu
115 120 125
Gly Asp Val Ala Thr Ala Phe Thr Met Ala Arg Ile Val Ser Asn Ile
130 135 140
Pro Ile Ser Arg Gly Gly Pro Thr Ser Val Val Ile Tyr Asp Ile His
145 150 155 160
Ala Leu Gln Glu Arg Phe Tyr Phe Ala Asp Gln Val Leu Pro Leu Phe
165 170 175
Glu Thr Gly Ile Pro Leu Leu Thr Lys Arg Leu Gln Gln Leu Pro Glu
180 185 190
Thr Glu Lys Val Ile Val Ala Phe Pro Asp Asp Gly Ala Trp Lys Arg
195 200 205
Phe His Lys Leu Leu Asp His Tyr Pro Thr Val Val Cys Thr Lys Val
210 215 220
Arg Glu Gly Asp Lys Arg Ile Val Arg Leu Lys Glu Gly Asn Pro Ala
225 230 235 240
Gly Cys His Val Val Ile Val Asp Asp Leu Val Gln Ser Gly Gly Thr
245 250 255
Leu Ile Glu Cys Gln Lys Val Leu Ala Ala His Gly Ala Val Lys Val
260 265 270
Ser Ala Tyr Val Thr His Gly Val Phe Pro Lys Ser Ser Trp Glu Arg
275 280 285
Phe Thr His Lys Lys Asn Gly Leu Glu Glu Ala Phe Ala Tyr Phe Trp
290 295 300
Ile Thr Asp Ser Cys Pro Gln Thr Val Lys Ala Ile Gly Asn Lys Ala
305 310 315 320
Pro Phe Glu Val Leu Ser Leu Ala Gly Ser Ile Ala Asp Ala Leu Gln
325 330 335
Ile
<210> 43
<211> 318
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）（菌株ATCC 204508 / S288c）（面包酵母）
<400> 43
Met Ser Thr Asn Ser Ile Lys Leu Leu Ala Gly Asn Ser His Pro Gly
1 5 10 15
Leu Ala Glu Leu Ile Ser Gln Arg Leu Gly Val Pro Leu Ser Lys Val
20 25 30
Gly Val Tyr Gln Tyr Ser Asn Lys Glu Thr Ser Val Thr Ile Gly Glu
35 40 45
Ser Ile Arg Asp Glu Asp Val Tyr Ile Ile Gln Thr Gly Tyr Gly Glu
50 55 60
His Glu Ile Asn Asp Phe Leu Met Glu Leu Leu Ile Leu Ile His Ala
65 70 75 80
Cys Lys Thr Ala Ser Val Arg Arg Ile Thr Ala Val Ile Pro Asn Phe
85 90 95
Pro Tyr Ala Arg Gln Asp Lys Lys Asp Lys Ser Arg Ala Pro Ile Thr
100 105 110
Ala Lys Leu Ile Ala Asn Leu Leu Glu Thr Ala Gly Cys Asp His Val
115 120 125
Ile Thr Met Asp Leu His Ala Ser Gln Ile Gln Gly Phe Phe His Ile
130 135 140
Pro Val Asp Asn Leu Tyr Gly Glu Pro Ser Val Leu Asn Tyr Ile Arg
145 150 155 160
Thr Lys Thr Asp Phe Asn Asn Ala Ile Leu Val Ser Pro Asp Ala Gly
165 170 175
Gly Ala Lys Arg Val Ala Ser Leu Ala Asp Lys Leu Asp Met Asn Phe
180 185 190
Ala Leu Ile His Lys Glu Arg Gln Lys Ala Asn Glu Val Ser Arg Met
195 200 205
Leu Leu Val Gly Asp Val Ala Gly Lys Ser Cys Leu Leu Ile Asp Asp
210 215 220
Met Ala Asp Thr Cys Gly Thr Leu Val Lys Ala Cys Asp Thr Leu Met
225 230 235 240
Asp His Gly Ala Lys Glu Val Ile Ala Ile Val Thr His Gly Ile Phe
245 250 255
Ser Gly Ser Ala Arg Glu Lys Leu Ile Asn Ser Arg Leu Ser Arg Ile
260 265 270
Val Cys Thr Asn Thr Val Pro Val Asp Leu Asp Leu Asp Ile Val Asp
275 280 285
Gln Val Asp Ile Ser Pro Thr Ile Ala Glu Ala Ile Arg Arg Leu His
290 295 300
Asn Gly Glu Ser Val Ser Tyr Leu Phe Thr His Ala Pro Val
305 310 315
<210> 44
<211> 427
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）（菌株ATCC 204508 / S288c）（面包酵母）
<400> 44
Met Arg Lys Cys Lys Ile Phe Val Gly Asn Ser His Pro Glu Leu Gly
1 5 10 15
Asn Met Val Cys Gln Arg Leu Gly Ile Glu Pro Ala Pro Cys Thr Leu
20 25 30
Lys Lys Phe Ala Asn Gly Glu Thr Ser Val Gln Ile Gly Val Ser Val
35 40 45
Arg Asp Glu Asp Val Tyr Val Ile Gln Ser Gly Ser Pro Ser Ile Asn
50 55 60
Asp Asp Ile Met Glu Leu Leu Ile Leu Val Ser Ala Cys Arg Gly Gly
65 70 75 80
Ser Ala Arg Lys Ile Thr Ala Val Ile Pro Gln Phe Pro Tyr Ser Lys
85 90 95
Gln Cys Lys Met Lys Arg His Arg Gly Ala Ile Thr Ala Arg Met Leu
100 105 110
Ala Asn Leu Leu Val Met Ala Gly Ala Asp His Val Val Ser Met Asp
115 120 125
Leu His Ala Ser Gln Met Gln Gly Phe Phe Thr Lys Pro Val Asp Asn
130 135 140
Leu Tyr Gly Gly Pro Ser Leu Ala Lys Trp Ile Arg Glu Asn Val Glu
145 150 155 160
Asp Tyr Glu Asp Ala Val Val Val Ser Lys Asn Pro Gly Gly Thr Lys
165 170 175
Arg Val Thr Ala Leu Ala Asp Ser Leu Lys Ile Asn Phe Ala Met Ile
180 185 190
His Thr Asp Arg Arg Arg Ser Lys Asp Leu Tyr Ser Gln Asn Lys Asp
195 200 205
Leu Gln Gln Leu Lys Leu Arg Lys Gln Ser Met Leu Arg Lys Asn Arg
210 215 220
Pro Ile Ile Arg Gln Gly Asp His Pro Asn Glu Glu Glu Asn Ile Ile
225 230 235 240
Leu Ser Asn Gly Ile Gln Thr Ala Arg Ile Arg Asn Gly His Val Ile
245 250 255
Gly Asp Asp Glu Ala Asp Asp Asp Glu Asp Ala Ile Leu Glu Ser Asp
260 265 270
Ser Glu Leu His Ser Ile Asp Gly Leu Asp Ser His Gly Leu Gly Gly
275 280 285
Thr Tyr Asp Ala Val Asp Ser Glu Asp Glu Glu Glu Ile Pro Val Leu
290 295 300
Tyr Arg Glu Gln Leu Ile Thr Leu Val Gly Asn Val Arg Gly Arg Ser
305 310 315 320
Ala Ile Ile Leu Asp Asp Met Ile Asp Arg Pro Gly Ser Phe Ile Ser
325 330 335
Ala Ala Glu His Leu Val Gln Asn Cys Gly Ala Lys Lys Val Tyr Val
340 345 350
Val Ala Thr His Gly Ile Phe Thr Gly Asp Cys Leu Glu Glu Leu Glu
355 360 365
Lys Ser Asp Ala Ile Asp Thr Ile Val Val Thr Asn Thr Tyr Pro Ile
370 375 380
Ser Gly Glu Arg Ile Ala Gly Ser Lys Lys Leu Val Thr Ile Asp Val
385 390 395 400
Ser Pro Ile Phe Ala Glu Cys Ile Arg Arg Asp His Tyr Gly Glu Ser
405 410 415
Ile Ser Val Leu Phe Asp Ser Leu Ala Ala Leu
420 425
<210> 45
<211> 320
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）（菌株ATCC 204508 / S288c）（面包酵母）
<400> 45
Met Pro Thr Asn Ser Ile Lys Leu Leu Ala Pro Asp Val His Arg Gly
1 5 10 15
Leu Ala Glu Leu Val Ala Lys Arg Leu Gly Leu Gln Leu Thr Ser Ser
20 25 30
Lys Leu Lys Arg Asp Pro Thr Gly Glu Val Ser Phe Ser Ile Gly Glu
35 40 45
Ser Val Arg Asp Gln Asp Ile Phe Ile Ile Thr Gln Ile Gly Ser Gly
50 55 60
Val Val Asn Asp Arg Val Leu Glu Leu Leu Ile Met Ile Asn Ala Ser
65 70 75 80
Lys Thr Ala Ser Ala Arg Arg Ile Thr Ala Ile Ile Pro Asn Phe Pro
85 90 95
Tyr Ala Arg Gln Asp Arg Lys Asp Lys Ser Arg Ala Pro Ile Thr Ala
100 105 110
Lys Leu Met Ala Asp Met Leu Thr Thr Ala Gly Cys Asp His Val Ile
115 120 125
Thr Met Asp Leu His Ala Ser Gln Ile Gln Gly Phe Phe Asp Val Pro
130 135 140
Val Asp Asn Leu Tyr Ala Glu Pro Ser Val Val Arg Tyr Ile Lys Glu
145 150 155 160
Asn Val Asn Tyr Met Asp Ser Ile Ile Ile Ser Pro Asp Ala Gly Gly
165 170 175
Ala Lys Arg Ala Ala Thr Leu Ala Asp Arg Leu Asp Leu Asn Phe Ala
180 185 190
Leu Ile His Lys Glu Arg Ala Arg Ala Asn Glu Val Ser Arg Met Val
195 200 205
Leu Val Gly Asp Val Thr Asp Lys Ile Cys Ile Ile Val Asp Asp Met
210 215 220
Ala Asp Thr Cys Gly Thr Leu Ala Lys Ala Ala Glu Ile Leu Leu Glu
225 230 235 240
Asn Arg Ala Lys Ser Val Ile Ala Ile Val Thr His Gly Val Leu Ser
245 250 255
Gly Arg Ala Ile Glu Asn Ile Asn Asn Ser Lys Leu Asp Arg Val Val
260 265 270
Cys Thr Asn Thr Val Pro Phe Glu Glu Lys Ile Lys Lys Cys Pro Lys
275 280 285
Leu Ala Val Ile Asp Ile Ser Ser Val Leu Ala Glu Ser Ile Arg Arg
290 295 300
Leu His Asn Gly Glu Ser Ile Ser Tyr Leu Phe Lys Asn Tyr Pro Leu
305 310 315 320
<210> 46
<211> 335
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）（菌株ATCC 204508 / S288c）（面包酵母）
<400> 46
Met Ser Glu Pro Ala Gln Lys Lys Gln Lys Val Ala Asn Asn Ser Leu
1 5 10 15
Glu Gln Leu Lys Ala Ser Gly Thr Val Val Val Ala Asp Thr Gly Asp
20 25 30
Phe Gly Ser Ile Ala Lys Phe Gln Pro Gln Asp Ser Thr Thr Asn Pro
35 40 45
Ser Leu Ile Leu Ala Ala Ala Lys Gln Pro Thr Tyr Ala Lys Leu Ile
50 55 60
Asp Val Ala Val Glu Tyr Gly Lys Lys His Gly Lys Thr Thr Glu Glu
65 70 75 80
Gln Val Glu Asn Ala Val Asp Arg Leu Leu Val Glu Phe Gly Lys Glu
85 90 95
Ile Leu Lys Ile Val Pro Gly Arg Val Ser Thr Glu Val Asp Ala Arg
100 105 110
Leu Ser Phe Asp Thr Gln Ala Thr Ile Glu Lys Ala Arg His Ile Ile
115 120 125
Lys Leu Phe Glu Gln Glu Gly Val Ser Lys Glu Arg Val Leu Ile Lys
130 135 140
Ile Ala Ser Thr Trp Glu Gly Ile Gln Ala Ala Lys Glu Leu Glu Glu
145 150 155 160
Lys Asp Gly Ile His Cys Asn Leu Thr Leu Leu Phe Ser Phe Val Gln
165 170 175
Ala Val Ala Cys Ala Glu Ala Gln Val Thr Leu Ile Ser Pro Phe Val
180 185 190
Gly Arg Ile Leu Asp Trp Tyr Lys Ser Ser Thr Gly Lys Asp Tyr Lys
195 200 205
Gly Glu Ala Asp Pro Gly Val Ile Ser Val Lys Lys Ile Tyr Asn Tyr
210 215 220
Tyr Lys Lys Tyr Gly Tyr Lys Thr Ile Val Met Gly Ala Ser Phe Arg
225 230 235 240
Ser Thr Asp Glu Ile Lys Asn Leu Ala Gly Val Asp Tyr Leu Thr Ile
245 250 255
Ser Pro Ala Leu Leu Asp Lys Leu Met Asn Ser Thr Glu Pro Phe Pro
260 265 270
Arg Val Leu Asp Pro Val Ser Ala Lys Lys Glu Ala Gly Asp Lys Ile
275 280 285
Ser Tyr Ile Ser Asp Glu Ser Lys Phe Arg Phe Asp Leu Asn Glu Asp
290 295 300
Ala Met Ala Thr Glu Lys Leu Ser Glu Gly Ile Arg Lys Phe Ser Ala
305 310 315 320
Asp Ile Val Thr Leu Phe Asp Leu Ile Glu Lys Lys Val Thr Ala
325 330 335
<210> 47
<211> 603
<212> PRT
<213> 酿酒酵母（Saccharomyces cerevisiae）（菌株ATCC 204508 / S288c）（面包酵母）
<400> 47
Met Pro Arg Asn Pro Leu Lys Lys Glu Tyr Trp Ala Asp Val Val Asp
1 5 10 15
Gly Phe Lys Pro Ala Thr Ser Pro Ala Phe Glu Asn Glu Lys Glu Ser
20 25 30
Thr Thr Phe Val Thr Glu Leu Thr Ser Lys Thr Asp Ser Ala Phe Pro
35 40 45
Leu Ser Ser Lys Asp Ser Pro Gly Ile Asn Gln Thr Thr Asn Asp Ile
50 55 60
Thr Ser Ser Asp Arg Phe Arg Arg Asn Glu Asp Thr Glu Gln Glu Asp
65 70 75 80
Ile Asn Asn Thr Asn Leu Ser Lys Asp Leu Ser Val Arg His Leu Leu
85 90 95
Thr Leu Ala Val Gly Gly Ala Ile Gly Thr Gly Leu Tyr Val Asn Thr
100 105 110
Gly Ala Ala Leu Ser Thr Gly Gly Pro Ala Ser Leu Val Ile Asp Trp
115 120 125
Val Ile Ile Ser Thr Cys Leu Phe Thr Val Ile Asn Ser Leu Gly Glu
130 135 140
Leu Ser Ala Ala Phe Pro Val Val Gly Gly Phe Asn Val Tyr Ser Met
145 150 155 160
Arg Phe Ile Glu Pro Ser Phe Ala Phe Ala Val Asn Leu Asn Tyr Leu
165 170 175
Ala Gln Trp Leu Val Leu Leu Pro Leu Glu Leu Val Ala Ala Ser Ile
180 185 190
Thr Ile Lys Tyr Trp Asn Asp Lys Ile Asn Ser Asp Ala Trp Val Ala
195 200 205
Ile Phe Tyr Ala Thr Ile Ala Leu Ala Asn Met Leu Asp Val Lys Ser
210 215 220
Phe Gly Glu Thr Glu Phe Val Leu Ser Met Ile Lys Ile Leu Ser Ile
225 230 235 240
Ile Gly Phe Thr Ile Leu Gly Ile Val Leu Ser Cys Gly Gly Gly Pro
245 250 255
His Gly Gly Tyr Ile Gly Gly Lys Tyr Trp His Asp Pro Gly Ala Phe
260 265 270
Val Gly His Ser Ser Gly Thr Gln Phe Lys Gly Leu Cys Ser Val Phe
275 280 285
Val Thr Ala Ala Phe Ser Tyr Ser Gly Ile Glu Met Thr Ala Val Ser
290 295 300
Ala Ala Glu Ser Lys Asn Pro Arg Glu Thr Ile Pro Lys Ala Ala Lys
305 310 315 320
Arg Thr Phe Trp Leu Ile Thr Ala Ser Tyr Val Thr Ile Leu Thr Leu
325 330 335
Ile Gly Cys Leu Val Pro Ser Asn Asp Pro Arg Leu Leu Asn Gly Ser
340 345 350
Ser Ser Val Asp Ala Ala Ser Ser Pro Leu Val Ile Ala Ile Glu Asn
355 360 365
Gly Gly Ile Lys Gly Leu Pro Ser Leu Met Asn Ala Ile Ile Leu Ile
370 375 380
Ala Val Val Ser Val Ala Asn Ser Ala Val Tyr Ala Cys Ser Arg Cys
385 390 395 400
Met Val Ala Met Ala His Ile Gly Asn Leu Pro Lys Phe Leu Asn Arg
405 410 415
Val Asp Lys Arg Gly Arg Pro Met Asn Ala Ile Leu Leu Thr Leu Phe
420 425 430
Phe Gly Leu Leu Ser Phe Val Ala Ala Ser Asp Lys Gln Ala Glu Val
435 440 445
Phe Thr Trp Leu Ser Ala Leu Ser Gly Leu Ser Thr Ile Phe Cys Trp
450 455 460
Met Ala Ile Asn Leu Ser His Ile Arg Phe Arg Gln Ala Met Lys Val
465 470 475 480
Gln Glu Arg Ser Leu Asp Glu Leu Pro Phe Ile Ser Gln Thr Gly Val
485 490 495
Lys Gly Ser Trp Tyr Gly Phe Ile Val Leu Phe Leu Val Leu Ile Ala
500 505 510
Ser Phe Trp Thr Ser Leu Phe Pro Leu Gly Gly Ser Gly Ala Ser Ala
515 520 525
Glu Ser Phe Phe Glu Gly Tyr Leu Ser Phe Pro Ile Leu Ile Val Cys
530 535 540
Tyr Val Gly His Lys Leu Tyr Thr Arg Asn Trp Thr Leu Met Val Lys
545 550 555 560
Leu Glu Asp Met Asp Leu Asp Thr Gly Arg Lys Gln Val Asp Leu Thr
565 570 575
Leu Arg Arg Glu Glu Met Arg Ile Glu Arg Glu Thr Leu Ala Lys Arg
580 585 590
Ser Phe Val Thr Arg Phe Leu His Phe Trp Cys
595 600
<210> 48
<211> 417
<212> PRT
<213> 粟酒裂殖酵母（Schizosaccharomyces pombe）（菌株972 / ATCC 24843）（裂殖酵母）
<400> 48
Met Ala Leu Leu Pro Phe Phe Asp Leu Thr Asn Phe Glu Ser Asp Ala
1 5 10 15
Ser Glu Glu Leu Gly Trp Leu Lys Tyr Val Gly Arg Val Gln Thr Arg
20 25 30
Val Phe Pro Gln His Phe Lys Asp Asn Leu Glu Lys Val Arg Lys Ile
35 40 45
Ser Glu Thr Ile Asp Val Ile Val Asp Thr Thr Ala Glu Leu Gly Pro
50 55 60
Glu Ala Cys Val Asn Leu Leu Asn Ala Gly Ala Leu Ala Ile Leu Val
65 70 75 80
Asn Glu Glu Met Leu Asn Glu Leu Ala Asp Ile Ser Pro Asn Arg Leu
85 90 95
Val Leu Lys Thr Asp Thr Thr Asp Ile Gly Lys Ile Glu Lys Leu Ser
100 105 110
Gln Val Ala Gly Ser Ile Gln Trp Ile Gly Ser Ala Glu Asn Tyr Pro
115 120 125
Pro Asp Phe Phe Glu Arg Ala Ser Lys Ile Ile His Lys Ala Val Met
130 135 140
Pro Glu Gly Gly Gly Arg Thr Leu Tyr Leu Glu Phe Pro Glu Gln Pro
145 150 155 160
Ser Met Glu Val Leu Lys Ser Phe Ser Val His Ser Val Val Pro Val
165 170 175
Leu Ser Ser Ser Phe Leu Thr Val Lys Pro Ala Glu Glu Pro Lys Lys
180 185 190
Leu Ser Leu Ala Asp Leu Ile Leu Ile Ser Ala Asn Thr Asp Arg Glu
195 200 205
Asp Gly Leu Phe Ser Thr Leu Val Val Asn Glu Leu Gly Ile Ala Leu
210 215 220
Gly Leu Val Tyr Ser Ser Lys Glu Ser Val Ala Glu Ser Leu Lys Thr
225 230 235 240
Gly Thr Gly Val Tyr Gln Ser Arg Lys Arg Gly Leu Trp Tyr Lys Gly
245 250 255
Ala Ser Ser Gly Ala Val Gln His Leu Ile His Ile Asp Val Asp Cys
260 265 270
Asp Glu Asp Cys Leu Arg Phe Val Val Tyr Gln Thr Gly Lys Gly Phe
275 280 285
Cys His Leu Asp Thr Leu His Cys Phe Gly Gln Ala Ser Gly Leu Cys
290 295 300
Gln Leu Glu Lys Thr Leu Ile Asp Arg Lys Asn Asn Ala Pro Glu Gly
305 310 315 320
Ser Tyr Thr Ala Arg Leu Phe Ser Asp Pro Lys Leu Leu Arg Ala Lys
325 330 335
Ile Met Glu Glu Ala Glu Glu Leu Cys Asp Ala Thr Thr Lys Glu Asn
340 345 350
Val Ile Trp Glu Met Ala Asp Leu Met Tyr Phe Ala Ile Thr Arg Cys
355 360 365
Val Gly Ser Gly Val Ser Leu Asn Asp Ile Ser Arg His Leu Asp Leu
370 375 380
Lys His Arg Lys Val Thr Arg Arg Lys Gly Asp Ala Lys Val Ala Trp
385 390 395 400
Gln Glu Lys Leu Lys Asp Lys Gly Gly Val Ala Asn Thr Ser Tyr Thr
405 410 415
Ala
<210> 49
<211> 316
<212> PRT
<213> 酒类酒球菌（Oenococcus oeni）（酒明串珠菌（Leuconostoc oenos））
<400> 49
Met Ser Glu Phe Asp Lys Lys Leu Asn Thr Leu Gly Val Asp Arg Ile
1 5 10 15
Ser Val Ser Pro Tyr Lys Lys Trp Ser Arg Gly Tyr Leu Glu Pro Gly
20 25 30
Asn Val Gly Asn Gly Tyr Val Ser Gly Leu Lys Val Asp Ala Gly Val
35 40 45
Ile Asp Lys Thr Asp Asp Met Ile Leu Asp Gly Ile Val Ser Tyr Asp
50 55 60
Arg Ala Glu Thr Lys Asn Ala Tyr Ile Gly Gln Ile Asn Met Thr Thr
65 70 75 80
Ala Ser Ser Phe Ser Gly Val Gly Gly Thr Val Leu Gly Tyr Asp Ile
85 90 95
Leu Arg Asn Pro Glu Val Asp Lys Ala Lys Pro Leu Phe Thr Glu Lys
100 105 110
Gln Trp Asp Gly Ser Glu Leu Pro Ile Tyr Asp Ala Lys Pro Leu Gln
115 120 125
Asp Thr Leu Val Glu Tyr Phe Gly Thr Lys Asp Asp Met Arg His Tyr
130 135 140
Pro Ala Pro Gly Ala Phe Val Cys Cys Ala Asn Lys Gly Val Thr Ala
145 150 155 160
Glu Arg Pro Lys Asn Asp Ala Asp Met Lys Pro Gly Gln Gly Tyr Gly
165 170 175
Val Trp Ser Ala Ile Ala Ile Ser Phe Ala Lys Asp Pro Thr Lys Tyr
180 185 190
Ser Ser Met Tyr Val Glu Asp Ala Gly Val Trp Glu Thr Pro Asn Glu
195 200 205
Asp Glu Leu Ile Glu Tyr Leu Lys Gly Arg Arg Asn Ala Met Ala Lys
210 215 220
Ser Ile Ala Ala Cys Gly Glu Asn Thr Ala Ala Glu Asn Gly Gly Ala
225 230 235 240
Val Phe Thr Ser Ser Trp Ile Gly Phe Ala His Ala Met Met Lys Pro
245 250 255
Gly Gln Val Gly Asn Ala Ile Thr Val Ala Pro Tyr Ile Ala Met Pro
260 265 270
Val Asp Ser Ile Pro Gly Gly Ser Ile Leu Thr Pro Asp Thr Asp Met
275 280 285
Asp Ile Met Gln Asn Leu Thr Met Pro Glu Trp Leu Asp Lys Met Gly
290 295 300
Tyr Gln Pro Leu Thr Lys Gly Gly Asn Ile Asn Tyr
305 310 315
<210> 50
<211> 283
<212> PRT
<213> 鸟乳杆菌鸟亚种（Lactobacillus aviarius subsp. aviarius）DSM 20655
<400> 50
Met Glu Pro Gly Asn Ile Gly Asn Gly Tyr Val Thr Gly Leu Lys Val
1 5 10 15
Asp Ala Gly Val Arg Asp Lys Thr Asp Asp Asp Val Leu Asp Gly Ile
20 25 30
Val Ser Tyr Asp Arg Ala Glu Thr Lys Asn Ala Tyr Ile Gly Gln Ile
35 40 45
Asn Met Thr Thr Ala Ser Ser Phe Thr Gly Pro Gln Gly His Val Val
50 55 60
Gly Tyr Asp Ile Leu Arg Asn Pro Glu Val Asp Lys Val Lys Pro Leu
65 70 75 80
Phe Val Glu Lys Gln Trp Asp Gly Ser Asp Leu Pro Ile Tyr Asp Ala
85 90 95
Lys Pro Leu Gln Asp Ala Leu Val Glu Tyr Phe Gly Met Glu Gln Glu
100 105 110
Arg Arg His Tyr Pro Ala Pro Gly Ser Phe Ile Val Cys Ala Asn Lys
115 120 125
Gly Val Thr Ala Glu Arg Pro Lys Ala Asp Glu Pro Leu Lys Pro Gly
130 135 140
Gln Gly Tyr Gly Val Trp Ser Ala Ile Ala Ile Ser Phe Ala Lys Asp
145 150 155 160
Pro Ser Lys Asn Ser Ser Met Phe Ile Glu Asp Ala Gly Val Trp Glu
165 170 175
Thr Pro Asn Glu Asp Glu Leu Ile Glu Tyr Leu Asn Gly Arg Arg Lys
180 185 190
Ala Ile Ala Lys Ser Ile Ala Glu Cys Gly Gln Asp Ala Asp Thr Ala
195 200 205
Phe Glu Ser Ser Trp Ile Gly Phe Ala His Val Met Met Lys Pro Gly
210 215 220
Gln Ile Gly Asn Ala Ile Thr Val Gly Pro Tyr Phe Ser Leu Pro Val
225 230 235 240
Asp Ser Ile Pro Asn Gly Ser Ile Leu Thr Pro Asp Lys Asp Met Glu
245 250 255
Ile Met Glu Asn Leu Ser Leu Pro Glu Trp Leu Asp Lys Met Gly Tyr
260 265 270
Glu Ser Leu Val Lys Lys Asn Asn Val Thr Tyr
275 280
<210> 51
<211> 387
<212> PRT
<213> 色杆菌属种（Chromobacterium sp.）LK1
<400> 51
Met Ser Leu Ser Pro Leu Asp Gln Asn Arg Ile Glu Ser Phe Trp Gln
1 5 10 15
Tyr Cys Leu Gln His Gln Tyr Phe Asn Leu Ala Tyr Pro Glu Ser Ala
20 25 30
Asp Phe Asp Tyr Ala Pro Leu His Arg Phe Leu Arg Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Asn Glu Ser Ser Asn Tyr Leu Leu Asn Ser Phe
50 55 60
Asp Phe Glu Arg Glu Val Met His Phe Phe Ala Glu Leu Phe His Ile
65 70 75 80
Pro Phe Asp Glu Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Ala Arg Glu Leu Phe Pro Asp Ala Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Ser His Tyr Ser Val Ala Lys Ile Ile Lys
115 120 125
Leu Leu Arg Ile Lys Ser Arg Ala Val Asp Ser Leu Pro Ser Gly Glu
130 135 140
Ile Asp Tyr Asp Asp Leu Val Ala Lys Ile Gln Gln Asp Gln Glu Arg
145 150 155 160
His Pro Ile Val Phe Val Asn Val Gly Thr Thr Met Lys Gly Ala Val
165 170 175
Asp Asp Ile Gly Val Ile Gln His Lys Leu Ala Glu Ala Gly Ile Pro
180 185 190
Arg Gln Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asp Ala Pro Gln Pro Tyr Ser Phe Ala Asp Gly Ile
210 215 220
Asp Ser Ile Ser Val Ser Gly His Lys Met Ile Gly Ser Pro Met Pro
225 230 235 240
Cys Gly Ile Val Leu Ala Lys Arg Ser Asn Val Ser Arg Ile Ser Val
245 250 255
Glu Ile Asp Tyr Ile Ser Ala Lys Asp Gln Thr Ile Ser Gly Ser Arg
260 265 270
Asn Gly His Thr Pro Met Met Leu Trp Ala Ala Ile Lys Ser Arg Pro
275 280 285
Leu Ala Glu Trp Arg Arg Lys Val Arg His Cys Leu Asp Met Ala Gln
290 295 300
Tyr Ala Ile Asp Arg Leu Gln Ala Ala Gly Ile Gln Ala Trp Arg Cys
305 310 315 320
Lys Asn Ser Ile Thr Val Val Phe Pro Ser Pro Ser Glu Pro Val Cys
325 330 335
Asp Lys His Gly Leu Ala Arg Ser Gly Gly Thr Ala His Leu Ile Thr
340 345 350
Thr Pro His His His Asp Ser Gln Arg Leu Asp Arg Leu Leu Asp Asp
355 360 365
Ile Val Gln Asp Leu Gly Ala Met Thr Ala Pro Ala Gly Ala Thr Met
370 375 380
Ser Ala Ala
385
<210> 52
<211> 1149
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 52
atgattctgt cccccgccga ccaagaaaga attgagacct tctggaatta ctgcctgaaa 60
catcagtact ttaacattgg ttaccctgag tctgccgact tcgattactc cgccctgttt 120
cggttcttca aattttctat taacaactgc ggcgactgga aggactactc caactacgcc 180
ctgaactcct ttgactttga gaaggacgtt atggcttatt tcgctgagat ctttcagatc 240
cccttcgaag agtcgtgggg atatgtcacc aatggtggca cagaaggcaa catgtttggt 300
tgttaccttg caagagagct ctttagcgat tcgaccctct actactctaa ggatacacac 360
tattccgtgg gcaagatcgc caagcttctt cagatgaaat cgtgcgtcat cgaatcgctc 420
gacaacggcg aaattgacta cgacgacctc attcacaaga tcaagacaaa taaggaatcg 480
catcctatca tctttgccaa cattggtaca actatgactg gcgctattga cgacattgag 540
atgatccagg agcgactcgc tcagatcgga attatgcgac gagattacta catccacgct 600
gacgcagccc tgtctggtat gattctgcct ttcgtcgacc atcctcaggc cttctctttt 660
gctcatggta tcgatagcat ctgcgtgtcg ggacataaga tgatcggatc ccctattcct 720
tgtggtattg tcgtcgctaa gcggcagaat gtggagcgaa tttccgtgga cgtggattac 780
atctctacac gagatcaaac tatttctggt tctcgaaacg gccataccgt gctgctgatg 840
tgggcagcca tccgttcgca gaccaacctc cagagacgac aacgaatcca gcactgtctg 900
aaaatggctc agtatgctgt cgatcgattt caagctgtcg gaatccctgc ctggcgaaac 960
cccaattcca tcactgtcgt ttttccctgt ccttctgagc acatttggaa gaagcactac 1020
ctggcaacat ctggcaacat ggctcacctt atcaccacag cccaccaccg agatacgcga 1080
cagattgact ctctcatcga tgacgtcatt tttgatcttc aaggtgcttc caagcgaacc 1140
gtcggcttc 1149
<210> 53
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 53
atggatctcg tcaaccatct gaccgaccgt ctcctgttcg ctatccccaa aaagggtcgg 60
ctctattcca agtctgtgtc gatcctcaac ggagctgaca ttacgtttca tcgatctcaa 120
cggctggata ttgctctctc cacgtccctg cctgtcgccc tgattttcct gcctgccgct 180
gacattccca ctttcgtggg cgagggtaag tgtgacctcg gtatcaccgg tgtcgatcag 240
gtgagagagt ctgacgtcga cgtggacctg gccatcgatc tgcagttcgg aaactgcaag 300
cttcaggtgc aggtgcctgt gaacggtgag tacaagaagc ctgagcagct gatcggtaag 360
accatcgtca catcctttgt taagctggca gagaaatact ttgctgacct ggagggcact 420
accgtcgaga agatgaccac ccgaatcaaa ttcgtttcgg gttctgtcga ggcttcctgc 480
gctctgggca tcggtgacgc cattgtcgac cttgttgagt ctggagagac catgagagcc 540
gctggactgg tggacattgc caccgttctg tcgacttcgg cctaccttat cgagtctaag 600
aaccccaaga gcgacaagtc cctgatcgca actattaaga gcagaattga gggtgttatg 660
acagcacagc ggtttgtgtc ctgcatctat aatgctcccg aggataaact tcccgaactg 720
ctcaaggtta cgcccggcag acgagccccc accatcagca agatcgacga tgagggttgg 780
gtcgccgttt cgtctatgat cgagagaaag acaaagggag tggtcctgga cgagcttaag 840
cgactgggtg cctctgacat tatggtcttt gagatttcca actgcagagt c 891
<210> 54
<211> 1149
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 54
atgattcttt caccggcgga tcaggaacgc attgagacct tctggaacta ctgcctgaaa 60
caccagtact tcaatatcgg ctatccagaa tcagcagatt tcgattacag cgcgctgttc 120
cgtttcttca aattttcgat aaataactgc ggagactgga aggactattc gaattacgcc 180
ttaaattcat ttgatttcga gaaagatgtg atggcctact tcgctgaaat ctttcaaatt 240
ccgttcgaag aatcatgggg ctatgtaaca aacggcggta ctgagggaaa catgtttggc 300
tgttatctgg caagagaact cttttccgat tcaaccctgt actattcaaa agatacacac 360
tattcagtcg gcaaaattgc gaaattactt caaatgaaaa gttgtgtcat tgagagttta 420
gataacggcg aaatcgacta tgatgattta atacataaaa ttaaaacaaa taaagaatca 480
catccaatca tatttgcgaa tatcggaact acaatgacgg gagctattga cgacatcgag 540
atgatacaag agagattagc acaaattgga attatgcgcc gcgattacta tattcacgca 600
gacgcggcct tgtcagggat gatacttcct tttgttgatc acccgcaggc attttctttc 660
gcgcatggta tcgatagcat ctgtgtttca ggacataaaa tgattgggag cccgatccca 720
tgtggtattg tcgtagcgaa aagacagaat gtcgaaagaa taagcgttga tgtggattac 780
attagcacac gcgaccagac aattagcggc tcacgtaacg gacatacagt cctgttaatg 840
tgggctgcta ttagatccca aactaattta caaagaagac aacgcatcca acattgcctt 900
aaaatggcac agtacgcggt agacagattt caagctgttg gaattccggc gtggagaaac 960
ccgaattcaa ttactgtagt ctttccgtgc ccttcggaac atatttggaa gaagcattat 1020
ttagcgacga gcgggaacat ggcccatctt atcacgacgg ctcaccacag agatacacgt 1080
caaattgatt ctttgattga tgatgtaatt ttcgatctgc aaggagccag taagagaacc 1140
gtgggattt 1149
<210> 55
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 55
atggatttag ttaatcactt gaccgataga ttattgtttg cgattcctaa aaaaggtcgg 60
ctttactcca aaagcgtgag cattttaaat ggcgcagata ttacctttca tagatctcag 120
cgtctggata tagcgctctc cacaagcctg ccggtggcgc ttatattctt accggcggca 180
gacatcccta ctttcgtggg ggaaggaaag tgcgacctcg gaatcacagg agtagatcaa 240
gttcgcgaat cggatgtaga tgttgacctt gccatcgatt tacaatttgg aaattgcaag 300
ctccaagttc aggtacctgt gaacggagaa tataagaagc cggaacagtt aattggcaaa 360
actattgtca cttcttttgt aaagctggcg gaaaaatatt tcgcggactt ggaaggtaca 420
accgtcgaaa aaatgactac acggatcaaa tttgtaagcg ggtctgttga agcaagctgc 480
gctctgggta ttggtgacgc gatagtggac ctggtggaaa gtggggaaac gatgcgagca 540
gcaggattgg tagatatcgc tacagtgtta tcaactagtg catacctgat agaatccaaa 600
aaccctaaat cggataaatc gcttatcgcc acaataaaat cacgcattga aggagtaatg 660
acggctcaac gatttgtgtc ctgcatatat aacgctcctg aggataaatt gccggagctg 720
ctgaaggtga caccgggccg gagagccccg accataagca aaatagacga cgagggctgg 780
gttgccgtaa gcagtatgat cgaacgcaag acgaagggag tagttttgga cgaacttaag 840
cgcttagggg catccgatat aatggtgttt gaaattagca actgtagagt c 891
<210> 56
<211> 1149
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 56
atgatcctat ctcctgctga ccaagaaaga attgaaacct tttggaacta ctgtttgaaa 60
catcaatact tcaacatcgg ttaccctgag agcgcagact ttgactatag tgcattattt 120
agatttttca aattctctat taacaactgc ggagactgga aagattactc caattacgct 180
ttaaactcct ttgattttga gaaggatgtg atggcatact ttgcagaaat attccaaatt 240
cctttcgaag aaagctgggg atacgttaca aatggtggta cagaaggtaa catgttcggt 300
tgttacttag cgagagaatt gttcagtgac tccacattgt attatagcaa agacactcat 360
tactcagtag gtaagattgc caaacttttg caaatgaagt catgcgtcat agaatcttta 420
gacaatgggg aaatagacta cgatgatctg atccataaga ttaaaactaa caaggaaagc 480
catccaatca tattcgcaaa tattggaaca acaatgacag gagcaatcga cgacatcgaa 540
atgatacaag aaagattagc ccaaataggc ataatgcgta gagactatta catacatgct 600
gacgcagcat tgagtggtat gattttgcct tttgtcgatc acccacaggc gttcagtttt 660
gctcatggta tcgatagtat atgtgtctcc ggacacaaga tgattggctc accgattcca 720
tgcggcattg ttgttgcaaa aagacagaac gtcgaaagaa ttagtgtgga cgtcgactat 780
atatcaacca gagaccaaac cattagcggt tcaagaaatg gccacacagt acttctgatg 840
tgggccgcga ttcgtagtca aacgaactta caaagaaggc aaagaattca acattgttta 900
aagatggcgc aatacgccgt cgacagattt caggctgtag gtatacctgc gtggagaaat 960
ccaaattcaa ttaccgtggt ttttccatgt ccttctgaac atatctggaa aaaacattac 1020
ttagccacta gcggtaacat ggcacaccta atcactaccg cccatcatag agatacaaga 1080
caaattgatt ccttaattga tgatgtcatt tttgacttac agggagcttc taagaggacc 1140
gttggtttc 1149
<210> 57
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 57
atggatttgg tcaaccacct tactgacagg cttttgttcg ccattccaaa gaaaggcagg 60
ttatactcca aatctgtttc aatactaaat ggagcagaca ttacttttca tcgtagccag 120
agactagata ttgctttatc aacctcactt ccagtagcac ttattttttt acccgcagca 180
gatataccaa catttgtagg tgagggtaag tgtgatttag gtattacagg ggttgaccaa 240
gttagagaaa gcgatgtcga tgtagacctt gctatcgact tgcagttcgg aaattgcaag 300
ttacaagtgc aggtgccagt taatggcgag tataagaagc cggaacagtt aataggaaaa 360
acaatagtta cctcttttgt gaagttagct gaaaaatact ttgccgactt agaaggtacg 420
actgtggaaa aaatgacaac aagaattaag tttgttagtg gttctgtcga agcatcctgc 480
gcactaggaa ttggtgatgc tattgtagac ttagtagaga gtggagaaac aatgagggcc 540
gcaggtttgg tagatattgc taccgtatta tcaacatccg catatcttat tgagtctaaa 600
aatcctaaat ccgacaaaag tttgatcgct actataaagt caaggattga gggtgtcatg 660
acagctcaga gatttgtatc atgcatctat aatgctcccg aagacaagct gcctgaatta 720
ctaaaggtaa caccgggaag aagggcacca actatcagca agattgatga tgaaggatgg 780
gtcgccgttt catcaatgat tgaaagaaag acaaaaggtg tggttcttga cgaacttaaa 840
cgtttgggag ccagtgatat catggtcttc gaaatttcta attgccgtgt g 891
<210> 58
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 58
atgctggaca acacaagact gcgaattgcc atccagaagt ctggtagact cagcgatgat 60
tccagagagc ttctggcccg ttgtggcatt aaaatcaacc ttcatactca gcggctcatc 120
gccatggccg aaaacatgcc tatcgatatt cttagagtcc gagatgatga catccctgga 180
ctggtcatgg acggcgtcgt cgatctgggt attattggcg aaaacgtgct cgaggaagag 240
ctcctcaacc gacgtgccca gggagaggat ccccgatacc tcaccctccg aagactcgac 300
ttcggaggct gccgactctc gcttgccacc cccgtcgacg aggcttggga cggacctgcc 360
gcccttgatg gtaagcgaat tgccacctct tacccccacc tgctcaagcg ttacctggat 420
cagaagggag tctcgttcaa gtcgtgcctg cttaacggta gcgtcgaggt ggctccccga 480
gctggcctcg ccgacgccat ttgtgatctg gtctccactg gcgccaccct cgaagccaat 540
ggtcttcggg aggttgaggt gatttaccga tccaaggctt gtctcattca gcgagatggc 600
gagatggccc aatccaaact gattgacaag ctcctgaccc gaattcaggg agtgatccag 660
gcccgagaat cgaagtacat catgatgcac gccccttccg aacgactcga ggaggtcatc 720
gccctgcttc ccggcgctga gcgtcccacc attcttcccc tggctggtga gcaacagcga 780
gttgccatgc acatggtttc ctcggaaact cttttttggg agacaatgga gaagctcaag 840
gccctcggcg cctcgtctat cctcgtcctt cctatcgaga agatgatgga g 891
<210> 59
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 59
atgttggata acacacgcct gagaatagcc attcagaagt ctggtagatt gtcagacgat 60
agtcgggaat tgctggctcg gtgtggaatt aagattaacc tgcatacgca gcgcctgatc 120
gcgatggctg aaaatatgcc aattgatatt cttcgtgtga gagacgatga tattccaggt 180
ttagtaatgg atggcgttgt cgatcttggc atcataggag aaaatgttct ggaggaggaa 240
ttattgaacc ggcgcgcaca gggcgaggat cctcgatatt taacattacg gcgactggac 300
tttggtggat gccggttgag cttagcgacg ccagtcgacg aggcttggga tggaccggct 360
gctttggatg gaaaacgtat tgcgacctca taccctcatc tgttaaaacg ctatcttgac 420
cagaaaggag tatcctttaa atcatgtctg ctgaatggat ctgtggaggt ggcccctcgt 480
gcaggattgg ccgatgccat ctgcgatctt gtgagcacag gtgcaactct tgaggcaaat 540
gggctcagag aagtcgaagt aatttaccgt agcaaagctt gcctgatcca gcgcgatgga 600
gaaatggcgc agtctaagtt aattgacaaa ttgctgactc gaatacaagg agttatccaa 660
gctagagaat ctaaatacat catgatgcat gctccatctg agcgactgga ggaagtaatt 720
gcgcttcttc ctggggcgga aagaccgact atcttgccgt tagcaggtga acaacagcgg 780
gttgccatgc acatggtttc ctctgaaact ctgttttggg aaaccatgga aaaactgaaa 840
gcgttgggag cgtcctctat ccttgttctt cctatcgaga aaatgatgga a 891
<210> 60
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 60
atgctggaca atacaagatt aagaatagca atacagaaat caggtagact gtcagacgat 60
agccgtgagc ttttagccag atgtggcata aaaattaatt tgcacacaca aaggcttatt 120
gccatggccg aaaatatgcc gatagacata ttaagagtga gagatgatga tatacccggg 180
ttggttatgg atggtgttgt cgatttgggt atcattggtg agaacgtgtt ggaagaagaa 240
ctgttaaaca gaagagctca gggagaggac cccagatatt taactttaag gcgtctggat 300
tttggtggtt gtcgtctttc cttagccaca ccagtagacg aagcttggga tggtccagca 360
gcactagacg gtaaaaggat tgcgacatcc tacccacatt tgctaaagcg ttatttggac 420
caaaaaggcg tttcttttaa gtcttgtcta ttaaatggct cagtagaagt ggctcctaga 480
gctggtctgg ctgatgcaat ctgtgattta gtgtctacag gtgcaacact agaagctaac 540
ggcttaagag aagtggaggt catttatagg tcaaaggctt gtttgatcca aagagacgga 600
gaaatggccc aatcaaaatt aattgataaa ctgcttacta gaattcaagg tgtgatccaa 660
gcaagagaat ccaaatatat catgatgcat gcaccgtcag agcgtctgga ggaggtgatc 720
gccttgttac caggtgcgga aagaccaacg attttgcccc ttgctggtga acaacaaaga 780
gtggccatgc acatggtatc ctctgaaacg ttattctggg agacgatgga aaaattaaag 840
gcattaggtg caagttctat attagtactg ccaatagaaa agatgatgga g 891
<210> 61
<211> 1149
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 61
atgatcctta gccctgccga ccaagaacgt atcgaaactt tctggaacta ctgcttgaaa 60
catcagtact ttaacatcgg atatccggag tctgccgact ttgactacag cgcacttttc 120
cgattcttca agttctcaat caacaattgc ggtgactgga aggattattc caactacgcc 180
ctgaactctt tcgatttcga aaaagatgta atggcatatt tcgcagagat cttccagatt 240
ccctttgaag aatcgtgggg ctacgtcacc aatggcggca ccgaaggcaa tatgttcgga 300
tgttacctgg ctcgcgaact cttttccgat tccacacttt attattcgaa ggatacccac 360
tactcggtgg gcaaaatcgc aaagctcctc cagatgaagt cctgtgttat cgaatctctt 420
gataacggtg aaatcgacta cgacgatttg attcataaaa tcaaaaccaa caaggagtct 480
catccaatta tttttgctaa cattggaacc accatgacgg gcgctattga cgatatcgaa 540
atgattcagg aacgcttggc ccagatcgga atcatgcgtc gagattacta tattcatgct 600
gatgccgcac tgtcgggtat gatcctccca ttcgtcgatc atccgcaagc cttttccttc 660
gcgcatggta tcgatagcat ctgtgtttcc ggccataaga tgattggctc gcccatcccc 720
tgcggcattg tggtggcaaa gcgccagaat gtcgagcgca tctccgtgga tgtggattat 780
atctctaccc gcgatcaaac catctcgggc tcccgcaacg gtcatactgt cttgctgatg 840
tgggcggcaa tccgttcaca aacgaacctc cagcgtcgcc agcgcatcca gcactgtctg 900
aaaatggcac aatacgcagt ggatcgcttt caagcagtcg gaattccagc atggcgcaac 960
ccgaactcca tcaccgttgt gttcccatgc ccgagcgaac acatctggaa gaagcactac 1020
ctggctacca gcggcaacat ggcccacctt atcaccactg cacatcaccg tgatactcgt 1080
cagatcgact cgctgatcga cgatgtgatc ttcgatctgc aaggtgcaag caaacgtact 1140
gtcggtttc 1149
<210> 62
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 62
atgttggata acacacgcct gcgcatcgct atccagaagt caggtcgtct gtcggacgac 60
tcccgcgagc tcctcgctcg ctgtggcatt aaaatcaacc tgcataccca acgcctgatt 120
gcgatggctg agaacatgcc tatcgacatc cttcgtgtcc gcgatgatga cattccaggt 180
cttgtgatgg atggcgttgt cgacttgggc atcattggcg aaaacgtgct cgaggaagaa 240
ttgctgaatc gccgagcgca aggcgaggac cctcgctatc tgaccctccg tcgactggat 300
tttggtggct gccgcctctc tttggcaact ccggtggacg aggcatggga tggtcctgcc 360
gcccttgatg gtaaacgcat cgccacttct tacccccact tgttgaaacg ctatttggat 420
caaaaaggcg tttcttttaa gagctgtttg cttaacggct cagtggaggt cgctcctcgc 480
gccggactgg cagatgctat ctgtgacctt gtttctaccg gtgctacctt ggaagccaac 540
ggattgcgcg aggttgaagt catctaccgc tccaaggcgt gcctcattca gcgagatggt 600
gaaatggccc agtccaagct tatcgataaa ctgttgactc gcattcaggg cgtgattcag 660
gcgcgcgaat cgaaatatat catgatgcac gcgccttctg aacgcctgga agaagtgatt 720
gcactgctcc ctggagccga gcgcccaacc atcctgcctc tcgcgggtga gcaacaacgt 780
gtcgccatgc acatggtctc gtctgaaaca ctgttctggg agaccatgga gaaacttaaa 840
gcactgggcg cgtcctcaat ccttgtcctg cccattgaaa agatgatgga a 891
<210> 63
<211> 1161
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 63
atgtccctgt ctcccctgga tcagaatcga attgagtcct tttggcagta ctgtctccag 60
caccaatact ttaacctcgc ttaccccgag tcggccgatt ttgactacgc ccctctgcac 120
cgattcctgc gattctccat caataactgc ggcgattgga acgagagctc caactacctc 180
cttaactcgt tcgacttcga gcgggaggtg atgcatttct tcgccgagct cttccatatt 240
cccttcgacg aaagctgggg ctacgtgacc aacggaggca ccgagggcaa tatgtttggt 300
tgctacctcg cccgagaact ctttcctgac gcaactctct actacagcaa ggacagccat 360
tactctgtgg ctaagatcat taagctgctg cgaatcaaaa gcagagcagt ggactccctg 420
ccctcgggtg agattgatta cgacgacctc gtggcaaaaa tccagcagga tcaggagcgg 480
caccctatcg tctttgtgaa cgtcggcaca accatgaagg gcgccgtcga tgacatcgga 540
gtcattcagc acaagcttgc tgaagccggc attccccggc aggactatta tctgcacgca 600
gatgctgctc tgtcgggcat gattcttccc tttgttgatg ccccccagcc ctacagcttt 660
gccgacggaa tcgactcgat ctccgtgtcg ggacacaaga tgatcggatc ccccatgccc 720
tgcggtattg tcctggctaa gcgttccaac gtctcgcgaa tttcggtcga gattgactac 780
atctctgcta aggaccagac catcagcggt tcccggaacg gtcacacccc catgatgctg 840
tgggccgcca tcaaatctcg acctctggct gagtggcgtc gtaaggtgcg acactgcctg 900
gatatggcac agtatgctat cgatcgactg caggccgccg gtattcaggc ttggcgatgc 960
aagaacagca ttacggttgt tttcccttcg ccatctgagc ccgtctgcga taagcatggc 1020
ctcgcacgat ctggtggtac agctcacctg attaccacgc ctcatcacca tgacagccaa 1080
cgactggacc ggcttctgga cgatatcgtt caggacctgg gagccatgac ggctcctgct 1140
ggcgctacca tgagcgccgc t 1161
<210> 64
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 64
atgctcaaga ttgcagtccc caacaagggt tcgctctccg agagagccat ggagattctg 60
gccgaggccg gctacgccgg cagaggagac tctaagtccc tcaacgtttt cgacgaggcc 120
aacaacgtcg agttcttctt cctccgaccc aaggatattg ctatctacgt tgccggtggc 180
cagctggatc tcggaatcac tggtagagac ctggcccgag attctcaagc cgatgtccac 240
gaagtcctgt ctctgggatt cggctcttcg acattccgat acgcagcccc tgctgacgag 300
gagtggtcga tcgagaagct cgatggcaag cgaatcgcca ccagctatcc aaatctggtt 360
cgagacgacc ttgcagcccg aggactctcg gctgaggtgc tgcgtctcga tggcgccgtc 420
gaggtctcca tcaagcttgg tgttgccgac gctattgctg acgtcgtctc cacgggccga 480
acactgcgac agcaaggtct tgcccctttc ggtgaggtgc tctgcacgtc cgaggccgtc 540
attgtcggac gaaaggatga gaaggtcact cctgaacaac agattctgct gcgtcgaatc 600
cagggtattc tgcatgctca gaacttcctc atgctcgatt acaaggtcga ccgggataac 660
ctcgatgccg ctactgccgt cactcccggc ttttctggac ctgccgtctc tcccctcgct 720
cgagacaact gggttgcagt cagagccatg gtgcctagaa gatctgctaa cgctattatg 780
gataagctcg ctggcctggg cgccgaagct atcctggcaa gcgagatcag aattgctcga 840
atc 843
<210> 65
<211> 1161
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 65
atgtctctaa gtcccttgga ccagaatagg atcgaatctt tttggcaata ttgcttgcag 60
caccaatatt tcaacctggc ctatcctgaa agtgctgatt tcgattacgc acctcttcac 120
agattcttga ggttctctat taataattgc ggtgactgga acgaatcttc aaactacctt 180
ttaaattcct ttgattttga aagggaagtt atgcacttct tcgccgaatt attccacatt 240
ccatttgatg aatcttgggg ctatgtcacc aacggtggta ctgaaggcaa tatgttcgga 300
tgttatcttg cacgtgaatt gtttccagat gctactcttt actactccaa agactcacat 360
tattcagttg ccaagattat taaactgctt aggattaaat ctagagctgt ggacagtctg 420
ccgagtggag aaattgacta cgacgacttg gtggcgaaga tacagcaaga tcaagaacgt 480
catccgatcg ttttcgtcaa cgtcggcaca acaatgaagg gagcagtgga cgatataggc 540
gtgattcagc acaaattggc tgaggcaggc atacctcgtc aagattacta tttacatgcc 600
gacgcggcac tttctggcat gatactacca tttgttgatg ctcctcaacc atattctttt 660
gctgatggaa tcgatagcat atctgtatca gggcataaga tgataggttc cccaatgccg 720
tgtggtattg tgttggcgaa gagatctaac gttagcagaa tatctgttga aatagattac 780
atttcagcta aagatcaaac aattagtggt agtagaaacg gccatacacc aatgatgttg 840
tgggcagcca taaaaagcag accgttagcc gagtggaggc gtaaggttcg tcattgcttg 900
gatatggctc agtatgctat agatcgtctt caagccgctg gcattcaagc ttggagatgt 960
aaaaactcca tcactgtcgt tttcccttct ccatccgaac ctgtctgtga caaacacggc 1020
ctagccagaa gtggtggtac tgcacacctg attactacgc cacatcacca tgattctcag 1080
agattagata ggctacttga tgatatcgtt caggatttgg gcgctatgac tgctccagct 1140
ggtgcgacaa tgagcgcagc t 1161
<210> 66
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 66
atgctgaaaa tagcggtgcc taataagggt tctttatcag aaagagcaat ggaaatacta 60
gctgaagctg gttacgccgg taggggtgat tctaaatcct tgaacgtgtt cgacgaagca 120
aataatgtag agtttttctt cttgaggccc aaagatattg ctatatatgt tgccggtggt 180
caattagatt taggtattac aggcagagat ttagctaggg attcccaggc agacgtgcat 240
gaagtcttat ctttaggatt cggtagttct acattcaggt acgcagctcc tgcagatgaa 300
gaatggagca ttgaaaaact agatgggaag agaatcgcaa catcctatcc taacctagtt 360
agagatgact tggccgcacg tggtctatca gccgaagttt tgcgtttaga tggtgcagtt 420
gaagtatcta ttaaattagg cgtggctgat gctattgcag atgtagtctc aacaggtcgt 480
acattaagac aacaaggatt ggctcccttc ggggaagttt tgtgtacctc agaagcagtc 540
attgttggca ggaaagatga aaaagttaca ccggaacagc aaattttatt aagaagaatt 600
cagggcatac tacatgcgca aaatttttta atgctagatt ataaggtgga tagagacaac 660
ttggatgcag ctaccgcagt aacacctggt ttttcaggcc cagcagtttc tcctctggcc 720
agagacaatt gggtcgctgt tagagcaatg gtgcctcgta ggagcgcgaa tgccataatg 780
gataagctag ctggtttagg agcagaagcc attttggctt ctgagatcag aatcgcaagg 840
att 843
<210> 67
<211> 1161
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 67
atgagccttt cccctttgga tcaaaatcgc attgagtctt tttggcaata ctgtttacag 60
caccaatact tcaaccttgc gtatccggag tccgcagatt ttgattatgc accgcttcac 120
cggtttctgc gtttttctat aaacaactgc ggggactgga atgaatcgtc taactatttg 180
ctcaattcat ttgacttcga acgtgaagtc atgcatttct tcgcagagct ttttcatatt 240
cctttcgacg agagttgggg gtacgtaacg aatggcggaa ccgaaggaaa catgtttgga 300
tgctaccttg cacgggaatt gtttcctgac gctacattat attacagcaa agatagccat 360
tatagcgtcg ctaaaatcat caaattgctt agaataaaaa gcagagcggt tgatagcctt 420
ccgtccggcg agatagatta cgatgatctt gtagccaaaa ttcagcaaga ccaagaacgg 480
catccaattg tttttgtaaa cgtcggaaca accatgaaag gtgccgtcga cgatattggt 540
gtcattcaac ataagctggc agaggcagga attccgcgtc aagattatta tctgcatgct 600
gacgcagcat tatcaggcat gatactgccg tttgtggacg ctccgcagcc gtactcattt 660
gctgacggta tcgattccat ttccgtctct ggccacaaaa tgattgggag cccgatgcca 720
tgtggtatag tactcgccaa acgctcaaac gtgagcagaa tttccgtcga aattgactat 780
atcagcgcga aggatcagac aattagtggt tctagaaatg gtcatacgcc gatgatgttg 840
tgggcagcga tcaaatccag acctttggct gaatggcgaa gaaaagtgcg tcattgtctg 900
gatatggctc aatatgctat agaccggctg caagctgcag gcatccaggc ctggcggtgt 960
aaaaactcaa ttactgtagt ctttccttcg ccgtccgaac cagtatgtga caaacatggg 1020
ttagccagat ctggcggcac ggcgcatctg attacaacgc ctcatcatca cgattcacaa 1080
cggttagatc ggcttttaga tgatatcgtg caggacctgg gtgcaatgac ggcaccggcg 1140
ggagccacaa tgagtgcagc c 1161
<210> 68
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 68
atgttaaaaa tagcagtacc taataaagga tctttgagcg aaagagctat ggaaatcttg 60
gcagaagccg gctatgcagg tcgtggcgat agcaaaagcc tgaatgtatt tgatgaagct 120
aataacgtcg agtttttctt tttgcggcca aaagatattg ctatttatgt ggcaggtggg 180
caactggacc ttggcatcac cggcagagac cttgctagag atagtcaggc cgatgtgcac 240
gaagtattaa gcttaggctt cggatcctct acttttcgct atgcggcccc tgctgatgaa 300
gaatggtcca ttgagaagct tgatggaaag cggatcgcga catcttatcc gaatcttgta 360
cgggatgatt tggcagctcg cggattgtct gcagaagtct taagacttga tggagcggtc 420
gaagtgtcta tcaagcttgg ggttgcggac gctattgcgg atgtagtctc tacaggccgt 480
acattgcgtc aacaagggct cgccccattc ggcgaagtgc tttgcacatc cgaagccgtg 540
atcgtaggca gaaaagatga aaaagttact ccggaacagc aaatactgtt gcgaagaatt 600
caagggatcc tgcatgctca gaatttttta atgttagact ataaagtcga tcgcgataac 660
ttagatgcgg caacggccgt tacgccgggg ttttcaggtc ctgctgtttc gccacttgct 720
cgcgataact gggttgcggt aagagctatg gttccgcgca gatcagctaa tgcaatcatg 780
gataagctgg ctggacttgg cgctgaagcc attctcgctt cagagattcg tattgcccgc 840
atc 843
<210> 69
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 69
atgtccgaat tggacgccaa acttaacaaa cttggggttg atagaattgc gatttccccg 60
tacaaacaat ggacgagagg atacatggaa ccggggaata tcggaaacgg ctacgttaca 120
ggacttaaag ttgatgcagg agttcgcgac aaaagtgacg atgacgttct tgatggaatc 180
gtcagttacg accgcgctga aacaaaaaac gcttatattg gacaaatcaa tatgacaaca 240
gcgagttctt ttacgggcgt gcaaggacgc gtaataggat atgatatctt gcgcagtcct 300
gaggtagaca aagcaaaacc gctgtttacg gaaacgcagt gggacggctc agagcttcct 360
atttatgatg caaaaccttt gcaagatgct ttggttgagt attttgggac agaacaagat 420
agaagacatt atcctgctcc gggatccttc atcgtgtgtg ccaataaagg agtgacagca 480
gaaagaccaa aaaatgatgc tgacatgaaa ccaggccagg ggtatggagt ttggtctgca 540
attgctatct cttttgccaa agatccgact aaagattctt cgatgtttgt agaggatgca 600
ggagtgtggg agacaccgaa tgaggatgag cttctggagt acctggaggg tagacgtaaa 660
gcgatggcta aatcaattgc ggaatgcggc caagatgcac atgctagctt tgaaagcagc 720
tggatcggtt ttgcatacac gatgatggaa ccgggccaga taggtaacgc gattactgtc 780
gcaccttatg tctcactgcc gatcgatagc atcccgggag gatcaatctt gaccccggac 840
aaagacatgg agattatgga gaatttaaca atgccggaat ggttagaaaa gatgggatat 900
aagtccctca gcgcgaataa cgccttgaag tat 933
<210> 70
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 70
atgtctgaac ttgatgcaaa actaaacaaa cttggggtcg atagaatcgc tatatctcca 60
tacaagcaat ggacccgtgg ttatatggaa cccggaaata taggtaatgg atatgtaact 120
gggctgaagg ttgatgccgg tgtaagagac aaaagcgatg acgacgtgct ggacggtata 180
gtctcttatg atagggccga gactaagaac gcttacatcg gccagattaa tatgactact 240
gcatcttctt ttacaggtgt gcaaggacgt gtaataggat acgatattct aagaagccca 300
gaagttgaca aggcaaaacc gctattcaca gaaactcaat gggacggctc tgaactaccg 360
atctacgatg ctaaacctct acaagatgct ttagttgaat acttcggtac ggagcaagat 420
aggcgtcact acccagcccc aggtagtttc attgtttgtg ctaataaggg tgttacagca 480
gaacgtccaa aaaacgatgc cgatatgaaa cctggtcaag gatatggggt ctggtctgcg 540
attgctattt ctttcgctaa ggacccaact aaagattctt ccatgttcgt tgaggacgca 600
ggcgtatggg agactccgaa tgaagatgaa ttgcttgaat acctagaagg taggcgtaag 660
gctatggcta agtcaatagc tgaatgtggt caagacgcgc atgcaagttt cgaatccagc 720
tggataggtt ttgcttacac aatgatggaa cccggtcaaa tcgggaacgc cattacggtc 780
gccccttacg taagtttgcc tattgacagt ataccaggag gttctattct tactcccgat 840
aaagatatgg aaattatgga aaacttaact atgccggagt ggttggaaaa gatggggtat 900
aaatctctgt ccgctaataa tgctttgaag tac 933
<210> 71
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 71
atgagcgaat tggacgcaaa actcaacaaa ctgggtgtgg atcgcatcgc aatcagccca 60
tacaagcagt ggacccgcgg ttatatggaa cccggtaaca tcggcaatgg ctacgtcact 120
ggacttaaag ttgacgcagg tgttcgtgat aaaagcgacg acgatgtgct ggatggaatc 180
gttagctatg accgagcaga gaccaaaaat gcatatatcg gacagatcaa tatgactacc 240
gctagctcct ttaccggagt gcagggacgt gtgattggct acgatatcct ccgctcgcct 300
gaggttgata aagccaaacc actcttcacc gaaacccagt gggatggctc cgaactgcca 360
atctacgatg cgaagcctct gcaggacgcc ctcgtcgaat attttggaac cgagcaggat 420
cgccgtcact atcccgcacc gggttccttc attgtgtgtg caaacaaagg cgtcaccgct 480
gagcgcccta aaaacgatgc agacatgaaa ccgggtcaag gatatggcgt ttggtccgcg 540
atcgccatct cgtttgccaa agacccaacc aaggactcat ccatgtttgt cgaggacgcc 600
ggagtgtggg agaccccaaa cgaggacgag ctcctggaat acctcgaggg ccgccgcaaa 660
gcgatggcaa agtcaattgc ggaatgcggt caggatgcgc acgcgtcttt tgaatcttcc 720
tggatcggat tcgcatacac gatgatggaa cccggtcaaa tcggcaacgc gatcactgtc 780
gccccttacg tctcactccc gattgacagc atcccgggtg gatcaatcct gaccccagac 840
aaggatatgg aaatcatgga gaatttgacg atgcccgagt ggttggaaaa gatgggctac 900
aagagcctgt ccgcgaacaa cgcgttgaag tat 933
<210> 72
<211> 1149
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 72
atgattctgt cacctgctga ccaagaacgt atcgaaacat tctggaatta ctgcttgaag 60
caccagtact tcaacattgg ttatcctgag tcggctgact tcgattactc cgccttgttt 120
cgtttcttca aattcagcat caacaattgt ggcgattgga aggattactc caactacgcg 180
ctgaactcgt tcgatttcga aaaggatgtc atggcctatt ttgctgagat tttccaaatc 240
ccgttcgagg aatcgtgggg ttacgtcacg aatggtggta ccgagggcaa tatgtttggc 300
tgttacctcg ctcgcgaact gttctccgat tccaccctgt actactccaa agatactcat 360
tattccgttg gcaagatcgc aaaactgttg cagatgaagt cctgcgtcat cgagtctctg 420
gataacggcg aaattgatta cgatgacctg attcataaga tcaaaaccaa taaggagtcc 480
cacccaatca ttttcgcgaa tattggtacg accatgaccg gtgctattga tgacattgag 540
atgatccaag aacgcctggc acagattgga atcatgcgac gcgattatta tattcatgca 600
gacgcggcgt tgtcaggcat gatcctccct tttgtggatc acccccaggc ctttagcttt 660
gcacatggca ttgactccat ctgcgtgtcg ggccacaaaa tgatcggctc accaatcccg 720
tgtggtatcg tggtcgcaaa acgccaaaat gttgagcgca tttcagttga tgtagattac 780
atctccaccc gagatcaaac tatctccggc tcccgaaacg gtcacactgt gcttctgatg 840
tgggcagcga tccgctcaca aaccaatctg cagcgccgtc agcgtatcca gcactgcctt 900
aaaatggctc agtacgctgt agaccgcttc caggctgtgg gtatcccagc atggcgcaac 960
ccaaactcca tcaccgtcgt cttcccatgc ccgagcgaac atatctggaa gaagcactac 1020
cttgccactt ccggtaacat ggctcatctg attactaccg cgcaccatcg agatacccgt 1080
caaatcgact ccctgattga cgatgttatt ttcgacttgc aaggcgcgtc aaaacgcacg 1140
gtcggcttc 1149
<210> 73
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 73
atggatctgg ttaaccactt gaccgatcgt ttgctctttg ccatcccgaa gaagggtcgc 60
ctctactcga agtcggtgtc cattctcaac ggcgccgata tcactttcca ccgctcccaa 120
cgactcgaca ttgcactgtc tacttctctg ccagtcgcac ttatcttctt gccagctgct 180
gacatcccaa cctttgttgg tgaaggtaaa tgtgacctgg gtatcactgg agttgatcag 240
gtgcgcgaga gcgacgtcga cgtcgatctt gcgatcgatc ttcaattcgg caactgcaag 300
cttcaggtac aagtcccggt caatggcgag tacaagaaac cagagcaact catcggcaag 360
accattgtca cttccttcgt taagcttgct gaaaagtatt tcgctgacct ggaaggaacc 420
acagtggaga aaatgaccac acgtattaag ttcgtttcgg gctcggttga ggcgtcctgc 480
gctttgggca ttggcgatgc aatcgttgat ttggtagagt ccggcgagac catgcgtgct 540
gcaggtctgg tcgatatcgc tactgttctg tccactagcg cttatttgat tgaaagcaag 600
aacccaaaga gcgacaagag cctgatcgca actatcaagt ctcgtatcga gggcgtgatg 660
acggcccagc gcttcgtgtc atgcatctac aacgcgcccg aagacaagct gcctgagctc 720
ttgaaagtga ccccaggccg tcgtgcgcct acaatttcca aaatcgacga cgagggttgg 780
gttgcagtgt cgtcaatgat cgaacgcaaa acaaagggtg tggtactgga tgaacttaag 840
cgcctcggag cctcagatat tatggtcttc gaaatcagca actgccgtgt t 891
<210> 74
<211> 404
<212> PRT
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 74
Met Thr Leu Ser Pro Ala Asp Gln Arg Lys Leu Glu Gly Phe Trp Gln
1 5 10 15
His Cys Val Thr His Gln Tyr Phe Asn Ile Gly Tyr Pro Glu Ser Ala
20 25 30
Asp Phe Asp Tyr Ser Gln Leu His Arg Phe Leu Gln Phe Ser Ile Asn
35 40 45
Asn Cys Gly Asp Trp Asn Glu Tyr Ser Asn Tyr Leu Leu Asn Ser Phe
50 55 60
Asp Phe Glu Lys Asp Val Met Thr Tyr Phe Ala Glu Leu Phe Asn Ile
65 70 75 80
Ala Leu Glu Asp Ser Trp Gly Tyr Val Thr Asn Gly Gly Thr Glu Gly
85 90 95
Asn Met Phe Gly Cys Tyr Leu Gly Arg Glu Leu Phe Pro Asp Gly Thr
100 105 110
Leu Tyr Tyr Ser Lys Asp Thr His Tyr Ser Val Ala Lys Ile Val Lys
115 120 125
Leu Leu Arg Ile Lys Cys Arg Ala Val Glu Ser Leu Pro Asn Gly Glu
130 135 140
Ile Asp Tyr Asp Asp Leu Met Ala Lys Ile Thr Ala Asp Gln Glu Arg
145 150 155 160
His Pro Ile Ile Phe Ala Asn Ile Gly Thr Thr Met Arg Gly Ala Val
165 170 175
Asp Asn Ile Val Thr Ile Gln Gln Arg Leu Gln Gln Ala Gly Ile Ala
180 185 190
Arg His Asp Tyr Tyr Leu His Ala Asp Ala Ala Leu Ser Gly Met Ile
195 200 205
Leu Pro Phe Val Asp His Pro Gln Pro Phe Ser Phe Ala Asp Gly Ile
210 215 220
Asp Ser Ile Cys Val Ser Gly His Lys Met Ile Gly Ser Pro Ile Pro
225 230 235 240
Cys Gly Ile Val Val Ala Lys Arg Asn Asn Val Ala Arg Ile Ser Val
245 250 255
Glu Val Asp Tyr Ile Arg Ala His Asp Lys Thr Ile Ser Gly Ser Arg
260 265 270
Asn Gly His Thr Pro Leu Met Met Trp Ala Ala Leu Arg Ser Tyr Ser
275 280 285
Trp Ala Glu Trp Arg His Arg Ile Lys His Ser Leu Asp Thr Ala Gln
290 295 300
Tyr Ala Val Asp Arg Phe Gln Ala Ser Gly Ile Asp Ala Trp Arg Asn
305 310 315 320
Glu Asn Ser Ile Thr Val Val Phe Pro Cys Pro Ser Glu Arg Ile Ala
325 330 335
Thr Lys Tyr Cys Leu Ala Thr Ser Gly Asn Ser Ala His Leu Ile Thr
340 345 350
Thr Pro His His His Asp Cys Ser Met Ile Asp Ala Leu Ile Asp Glu
355 360 365
Val Val Ala Glu Ala Gln Leu Asn Thr Leu Arg Ser Lys Arg Ala Phe
370 375 380
Thr Asp Gln Thr Val Val Glu Arg Leu Pro Ala Ala Ser Phe Asn Leu
385 390 395 400
Arg Thr His Tyr
<210> 75
<211> 1002
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 75
atgcagaaat atgaatacga tcctaagtgg gtaatcaata atgcgatttc aagcgagcgt 60
gagttttgca ccggctacca aaaccccggc gcgtctggaa atggctatgt cacgactatt 120
aagctttcta cgggtctcgt ggacattacc ccttgggaaa aagtccaggc catgagcgag 180
catgaggtta ttaagttcga tcgcggttgc tctaatattg ttagctatga tcgttgtgaa 240
tgtaatgacg cctatattgg cgcaatcaac atgttgactg cctcctcctt ctctggtctg 300
caaggcgtca tttggggtta cgatatcgct gtagtggaga acctgcggag ccggaagctt 360
tacgatcaaa agtggccgag cggcgacccg aattcggaat attccacgcc ggtttattct 420
attgagccgc tgctcaacgc aacggagcgc cttttcggcc atgcggagcc tggcaagcgt 480
cgctttaatc ccatccccgg atccatggtg gtctgtgcaa ataaatctgc cacttcagac 540
cctagctcag atgtaaaaga gggctgggct ttttctgtca tctcccttgc aattcttgag 600
aaccggaata gcggatcaaa tctcttcatt gaagactgcg acattattga tatcaataat 660
ccagacggta cccgtaagac taaagaggat gtcaaagcga tgctcgacac gacccttcgc 720
aaggtcacgg aatgcacggt attgtgtggt ctcgaccaac acatcaagta caaagagatt 780
ttcatcggat acaaggtgat taagttcaac gagaagcaag taggttgtgc gctcgcgtgt 840
gcaccgtacg tgacgcttgc acggaatgca gtgcctcagg gaatgaaacc gtcgaaactg 900
acggacatga acatttctca atgggagaac gctctgaatc tgcaacccct ggagaagatt 960
gaaaaatcaa agatcggcat tttgggaatg ggtgtgcttg ac 1002
<210> 76
<211> 1047
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 76
atgactctgc actcacggta ttctaaattg aaaggtgaga acaaaatgca aaagtacgag 60
tacgacccaa agtgggttat caacaacgca atctcgtccg aacgggaatt ttgtaccggc 120
tatcagaacc caggtgctag cggcaacggc tatgttacta ctattaaact gtcaacgggc 180
ttggtagata ttacgccatg ggagaaggtg caggcaatgt ccgagcacga agtgattaag 240
ttcgaccggg gatgcagcaa cattgtctct tacgaccggt gtgaatgtaa tgatgcttac 300
attggagcca ttaacatgct gactgcttct tctttctcgg gactgcaggg agtcatctgg 360
ggttatgata tcgcagtcgt ggaaaatctg cgttcgcgga agctctacga tcaaaaatgg 420
ccttccggcg atccaaattc ggaatattct acgcctgtgt attcgattga gccgttgttg 480
aatgccactg aacggctctt cggtcacgcg gagccaggaa agcgccgttt taacccaatc 540
cctggatcaa tggtggtgtg tgcgaacaag tcagctacct cggatccgtc ttcggacgtt 600
aaggagggat gggcattttc agttattagc ctcgctatcc ttgagaatcg caactctggt 660
agcaatctgt ttatcgaaga ctgtgatatt atcgacatta acaatcccga tggtactcgc 720
aaaaccaaag aggatgtgaa ggctatgttg gataccactt tgcgcaaagt tacggagtgt 780
acggttctct gcggtcttga ccagcatatt aagtataaag aaatcttcat tggctataaa 840
gtcattaaat tcaacgagaa gcaagtggga tgcgctcttg cctgcgcgcc ctatgttact 900
ctcgcacgta atgctgtacc gcaaggaatg aaaccgtcca agctcactga tatgaacatt 960
tcacagtggg aaaatgcact gaacttgcag ccactggaaa agattgaaaa gtcgaaaatc 1020
ggcattcttg gaatgggagt gctcgat 1047
<210> 77
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 77
atgagcgaac tcgacgcaaa actgaataaa ctcggagtag accggatcgc gatttccccg 60
tataagcaat ggacgcgggg atacatggag ccaggtaaca tcggaaatgg ttacgtgacg 120
ggacttaagg tagatgctgg tgtacgcgat aaatccgatg atgatgtatt ggacggtatt 180
gtcagctatg accgggcgga gacgaaaaac gcctacatcg gccaaatcaa catgaccact 240
gcatcgtcat ttactggcgt ccagggccgg gtgattggat acgacatcct tcgctcccct 300
gaggtagaca aggcgaagcc gctgttcacg gagacgcaat gggatggttc agaacttccg 360
atttatgacg cgaaaccgct tcaagacgct cttgtagagt attttggcac cgagcaagat 420
cgtcgccatt acccagcacc aggatccttc attgtttgtg ctaataaggg tgtaaccgca 480
gagcggccta agaacgacgc agacatgaag cctggccagg gctacggcgt atggtcagcg 540
attgccatta gctttgcaaa ggatcccacc aaagattctt caatgtttgt agaagatgca 600
ggagtctggg agactccgaa cgaagatgag ctcctggaat accttgaggg acggcgtaag 660
gcgatggcaa agtctatcgc ggagtgtggt caggacgcac atgcgtcctt tgaatcgtcg 720
tggattggct tcgcgtatac gatgatggag cccggccaaa tcggcaacgc tattactgtg 780
gccccctatg tttctttgcc cattgacagc atcccgggag gaagcatcct taccccggat 840
aaagatatgg aaattatgga gaacctcacg atgcccgaat ggctggagaa aatgggatac 900
aagtctctca gcgctaacaa cgcacttaaa tat 933
<210> 78
<211> 1149
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 78
atgattttgt ctccagccga ccaggaacgt atcgagactt tctggaatta ttgcctgaaa 60
catcagtact ttaacatcgg ctaccccgaa tcagccgact ttgactactc tgcactgttc 120
cgtttcttta agttttccat taacaactgc ggagactgga aggactattc aaactatgcc 180
cttaacagct tcgacttcga gaaagacgtc atggcgtact ttgctgagat ttttcaaatc 240
ccctttgaag agtcttgggg atacgtgact aatggcggca ctgagggcaa tatgtttggt 300
tgctacctcg cccgtgagtt gttctccgat tctacgcttt attattcgaa ggacacgcat 360
tatagcgtcg gcaaaatcgc caagctgctc caaatgaaat cctgcgttat cgagtccctc 420
gacaatggcg aaatcgacta tgacgacttg attcataaaa ttaaaacgaa taaggagtcc 480
cacccgatta ttttcgctaa tatcggtacc actatgacgg gtgcaattga tgatattgaa 540
atgattcaag aacgtctggc tcagatcggc attatgcggc gggactacta tattcacgcg 600
gacgcagctc tttctggcat gatccttccc ttcgtcgatc atccacaagc gttttccttc 660
gcacatggca tcgattcaat ttgtgtttcg ggccataaga tgattggaag cccaattcca 720
tgtggcattg tcgtagctaa acgccaaaac gtggaacgta tctcagtaga tgtggattat 780
atcagcactc gtgaccaaac catctcgggt tcccggaatg gacacaccgt gttgctcatg 840
tgggcggcaa tccggtcgca gaccaatttg caacgtcggc agcggatcca acattgcctg 900
aagatggctc agtacgcagt cgatcggttt caagcagtag gtattcctgc atggcggaac 960
ccgaatagca tcacggttgt gttcccatgt ccttctgaac acatctggaa aaaacactat 1020
ctcgcgactt cgggcaatat ggctcatctc atcactaccg ctcatcatcg cgatactcgt 1080
cagattgatt cgctgatcga cgacgtgatc tttgatttgc agggtgcgtc caaacggacc 1140
gtgggattt 1149
<210> 79
<211> 2541
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 79
atggatttca aggagtaccg ccagcgcgga aaggagatgg tcgattatat tgccgactac 60
cttgagaata ttcgcgagcg gcgcgtcttc cccgatgtaa gcccgggcta tatgcgtcag 120
ctcttgcccg aatcagcgcc gatcgaagga gagccgtggc cgaagatctt cagcgatgta 180
gagcgcattg taatgccagg aatcacgcac tggcagtccc cgcatatgca tgcctatttc 240
ccagccttga attcaatgcc ctccctcctc ggagatatgt tggccgatgc gattaactgt 300
ctgggcttca cctgggcatc aagccccgca tgcactgagt tggagatcat cgtcatgaat 360
tggttgggca agatgattgg cctgcccgac gcattcctgc acctgtcgtc ccaatcacaa 420
ggtggtggcg tcctccagac tactgctagc gaggccactt tggtgtgcct gctggcagga 480
cggactcgtg caatccaacg tttccatgag cggcatcctg gttaccaaga cgcggaaatt 540
aacgcgcggc ttgttgctta ttgctccgac caagcacatt cgtcggtgga gaaggcagct 600
ttgatcggac ttgtccgtat gcgctatatc gaggctgacg atgacttggc catgcgtggt 660
aagctgctgc gggaggcgat tgaggacgat attaaacaag gattggtgcc attctgggtc 720
tgcgctactc ttggcaccac cggttcttgt agctttgaca atttggagga aattggaatt 780
gtttgcgctg agcaccactt gtggcttcac gtagatgcgg catacgcggg tagcgctttc 840
atttgccctg agtttcggac gtggttgcgg ggcatcgaac gtgccgactc gattgctttc 900
aacccctcga aatggctgat ggtccacttt gacgccactg ctctttgggt gcgggatagc 960
accgcggtcc atcgcacttt caacgtcgaa cccctctacc tccaacatga aaactcaggt 1020
gttgctgttg atttcatgca ttggcaaatc cccctgtcgc ggcggtttcg tgctttgaaa 1080
gtgtggttcg tattgcgctc ttacggtatt aaaggactgc aacggcatat ccgcgaaggt 1140
gttcgcctgg cgcagaaatt tgaagccctc gtcctggccg atcaccgttt cgagctgcca 1200
gcaaaacgcc accttggcct ggtggtattc cgtatccgcg gcgacaatga gatcactgaa 1260
aaattgctta agcgtcttaa ccaccgtggc aacctccact gcatcccatc gtccctcaag 1320
ggtcaatatg ttattcgctt caccatcacc tcaacccaca cgacgctcga tgatattgta 1380
aaggattgga tggagatccg ccaggtagcg tccacggtac tggaggaaat gaatattact 1440
atttcgaatc gtgtttacct gaaggaaacg aaagagaaga acgaagcctt cggatcgtct 1500
ctgctgctct ctaactcacc tctgtctccg aaagtggtca acggttcctt cgcagcaatc 1560
ttcgacgcgg acgaattcct cgctaaaacg tatgccggcg tgcgtattgc tcatcaggag 1620
tcaccctcca tgcgccgtcg tgtgcgcggc attctgatga gcggcaagca gttctccctg 1680
gattcccaca tggacgtcgt ggtgcaaacg acgctcgacg ctggaaacgg agcaacgcgt 1740
acttctacta ctaactcgta tggtcacacc acttccgccg cacaggccaa ttcagagcgg 1800
caggcttcta tccaggaaga taacgaggaa agcccagagg agactgaact tttgtcgctt 1860
tgccgtacct caaacgttcc cagccccgaa catgcacact ccttgtctac gccatcccgc 1920
tcatgctcgt ccagctctca ttcgttgatc cattcactta cccagtcgtc tccgcggtca 1980
agcccggtta atcagtttcg gcctattacc ctctgcgccg taccctccca atcccagctg 2040
tccatgccgc ttgcaatgcc tttgccgaac cgcaacgtca ctgtttcggt agattcactg 2100
ctcaacccag ttacgacttg caatgtgtac cacggcaaac gttttctcga acccctcgag 2160
aacttggccc aaacctctgc atctttctcc tcttctattt tccggctgcc cacccccatt 2220
gccactccca ctcgggagag cccggaggat cccgactggc cagcgaagac tttctctcaa 2280
ctcttgttgg aacggtatag cagccaaagc caatcgctgg gaaataattc ctctacggaa 2340
tcaagcagct tgtccggcgg tgccaccccg acgccgacgc caatgtcgtc acttgacgaa 2400
ctcgtaacgc ctcttctgtt gtctttcgct tccccaagcc agcccatgct ctcggcacat 2460
ggtatcggtg aaggacaacg cgagcaggga tcggactcag acgcgacggt ttgctccacg 2520
acttcctcta tggaatcact g 2541
<210> 80
<211> 1161
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 80
atgtccctgt cccctttgga tcaaaatcgc attgagtcct tctggcaata ttgtttgcag 60
catcagtact tcaatctggc ttatcccgaa tctgcggatt tcgactacgc ccccttgcat 120
cgtttcctcc ggttctctat caacaactgc ggagattgga acgagtcctc aaattacctg 180
cttaactcgt ttgatttcga gcgggaggta atgcacttct tcgcggaatt gtttcatatt 240
ccctttgatg agtcttgggg ttatgttact aatggaggta ctgaaggcaa tatgtttggc 300
tgctaccttg ctcgcgagtt gttccctgat gcaaccttgt attactcaaa agatagccat 360
tactccgtcg ctaagattat caaattgttg cggatcaaat cgcgcgctgt ggacagcctc 420
cctagcggtg aaatcgatta tgacgatctt gtggcaaaga ttcaacagga tcaggaacgt 480
cacccaatcg tttttgttaa cgtgggtacg actatgaaag gcgctgtgga cgatattgga 540
gttattcagc acaaacttgc tgaagcaggt atcccccggc aagattacta cttgcatgcg 600
gatgcggcac tttccggcat gatcctcccg tttgtggatg caccccaacc ctattccttt 660
gctgatggaa ttgacagcat ctccgtatct ggacataaga tgattggaag cccgatgcct 720
tgtggcattg ttttggcaaa acggtccaat gtctctcgga ttagcgtaga gatcgattac 780
atctccgcaa aggaccagac gatctctgga tcccgtaatg gtcacacccc tatgatgctc 840
tgggctgcaa tcaaatcacg gccgcttgcg gaatggcgtc ggaaggttcg ccattgtttg 900
gatatggctc agtacgcgat tgatcgtctc caagctgcag gtattcaagc gtggcgctgc 960
aagaacagca ttacggttgt atttccttcg ccctcagagc ctgtgtgcga caagcatggc 1020
ttggcccgct ctggcggtac cgcgcacctc attaccactc cgcatcacca tgactcccaa 1080
cgcttggacc gtctcttgga cgacatcgtc caagacttgg gcgcaatgac ggctcctgct 1140
ggtgcaacta tgtctgccgc a 1161
<210> 81
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 81
atgttggaca atacgcgcct ccgcatcgcc atccaaaaaa gcggccgtct gtcagacgac 60
tcgcgtgaac tgctcgcccg gtgcggtatc aagattaacc ttcacaccca acgtctgatt 120
gcaatggcgg aaaacatgcc tattgacatt ttgcgtgtcc gggacgatga cattcctggc 180
ctggtaatgg acggcgtcgt tgatctggga attattggtg agaatgtgct tgaggaggag 240
ctcctgaacc ggcgcgcgca gggtgaggac cctcgctacc tgactctgcg ccgcctggat 300
tttggcggat gtcggctgtc gcttgcaacg cccgtggatg aagcctggga tggacctgct 360
gcgctggacg gcaagcgtat tgctacctca tatcctcatc ttttgaagcg ttatttggat 420
caaaaaggtg tatccttcaa gagctgcctt cttaacggtt cagtggaggt cgccccgcgc 480
gccggtcttg ctgatgctat ttgtgacctc gtttcgacgg gtgccaccct tgaggcgaac 540
ggtctgcgcg aagtggaagt aatttatcgc tcgaaagcgt gtcttatcca gcgggacgga 600
gagatggcgc agtccaaact tatcgataaa ctgcttactc gtattcaggg tgtgatccaa 660
gcccgggaaa gcaagtatat catgatgcac gccccgtcgg aacgcctgga agaagtcatt 720
gcgcttcttc cgggagctga acgtccaacc atcctgccgc tggcaggcga acaacaacgt 780
gttgctatgc acatggtttc aagcgaaact ctgttctggg agacgatgga aaagcttaag 840
gctcttggcg cgtccagcat tttggtgttg ccgattgaaa aaatgatgga a 891
<210> 82
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 82
atgtccgagt tagacgcaaa gctaaataaa ctgggtgtcg acagaatcgc aatttctcca 60
tataagcaat ggacgagagg gtatatggag ccagggaata tcgggaacgg ctacgtcact 120
ggacttaaag tagacgccgg ggttagagac aaaagcgatg atgacgtcct agatggaata 180
gtatcctacg atagagctga aaccaagaac gcatacattg ggcagataaa tatgacgact 240
gctagctctt ttacgggtgt acagggtagg gttattggat acgacatctt gagatccccg 300
gaagtcgata aggccaaacc gttatttaca gaaacgcagt gggatgggtc agagctaccg 360
atatatgacg caaaaccatt gcaagatgcg ttagtggagt acttcgggac cgaacaagat 420
agaagacact accctgctcc cggttcattc atagtatgcg ccaataaagg cgtaacagcc 480
gaaaggccga aaaacgatgc ggacatgaaa ccaggtcaag gttacggtgt gtggtctgct 540
atagcgatta gttttgcgaa agatcctacc aaagatagca gcatgttcgt cgaagacgct 600
ggtgtgtggg agacccctaa cgaggacgaa ttacttgagt atttggaagg cagaagaaaa 660
gcgatggcta agtcaattgc tgaatgcggg caggacgcac acgcgtcctt tgaaagcagc 720
tggataggct ttgcatatac aatgatggag ccaggtcaaa taggtaacgc gatcaccgta 780
gcaccttacg tgtcccttcc tatcgattcc atccctggag gtagcatttt gacgcctgac 840
aaggatatgg agatcatgga gaaccttaca atgcctgagt ggctagagaa gatggggtac 900
aaatccctat ccgcgaacaa cgcgctaaag tac 933
<210> 83
<211> 1239
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 83
atggaatcag acattaagaa cgagacctct ttccaagagc ttgatatgat cctgacacag 60
tacctggaaa cactatcaga aagaaaaaag tatcatatag gctaccccat aaacatgtgt 120
tacgagcatc atgcgaccct ggcacccttg cttcagtttc acctgaataa ctgcggtgat 180
ccgttcacac aacaccccac tgactttcac agcaaggact ttgaggttgc ggtgcttgac 240
tggttcgccc aactgtggga gatagagaag gacgagtact ggggttatat aacttccgga 300
ggaactgaag ggaatctgca tggtttttgg ttaggtagaa gagagttgtt gccaaatggt 360
tacctatatg ccagcaaaga ttcccattac tccatattta aagccgctag aatgtacaga 420
atggagttac aaactatcaa tacgctagtt aatggagaga ttgattacga agacttgcag 480
agtaaactgc ttgtcaacaa gaacaagccc gcgataatta atatcaatat tgggacaact 540
tttaaaggtg ctatagatga cttggatttc gttatacaga cattagaaaa ctgcggctat 600
tctaatgata actattacat tcactgcgac agggctctat gtggactaat tcttccattt 660
atcaaacacg cgaagaagat cacgttcaag aagcccattg ggagcatcag catttctggg 720
cacaagttct taggttgccc gatgagctgc ggcgtgcaaa taacacgtag gtcatatgta 780
agtacactga gcaaaatcga gtacattaac tccgcggacg caaccattag tggcagccgt 840
aacggattca cgccgatctt tctgtggtat tgtctatcta agaaaggaca tgcgagatta 900
caacaagatt ctattacctg tattgagaat gcgagatacc taaaggatag gctgctggaa 960
gccgggatta gcgtaatgct gaatgatttt agcatcacag tagtattcga acgtccctgt 1020
gatcataaat ttattagaag atggaacctg tgctgtctgc gtggcatggc tcacgtagtc 1080
atcatgccgg ggataactcg tgagacgatt gatagttttt tcaaggatct gatgcaggag 1140
cgtaattata aatggtacca ggacgtgaaa gccctaccac cttgtttggc agatgactta 1200
gcattgaact gcatgtgtag caataagaaa atgcataac 1239
<210> 84
<211> 1986
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 84
atgatggagc cgtgtgaata cagggagtac agggagtatt acagagcgag gggtaaagag 60
atggttgatt atatcagcca gtatttaagt acagtcaggg agagacaggt cacccctaat 120
gtacagcccg ggtatctacg tgcgcagttg ccagcatccg caccagaaga gcctgattct 180
tgggattcta tctttgggga tatagagaga gtcattatgc ctggggtcgt ccattggcaa 240
tctccgcaca tgcatgctta ctatcccgcg ctgactagtt ggccgtccct acttggagat 300
atgttagctg atgcgattaa ctgtcttggc ttcacctggg ccagctcacc cgcttgcact 360
gaactggaaa tgaatattat ggactggtta gcgaagatgt tagggcttcc ggaatatttt 420
cttcaccatc acccatcatc cagaggagga ggggtcttac aatccacagt ctctgagtcc 480
acgctgatag cacttttagc ggctcgtaaa aataaaatct tagccatgaa ggcttgcgaa 540
cccgatgcaa atgagagttc actaaacgca aggctggtag cgtatacctc agatcaagcg 600
cattcctcag ttgaaaaggc cgggcttatt agcttggtta agatcaggtt cctaccggtt 660
gatgataatt ttagtctaag gggggaagcg ctgcaaaagg ctatcgaaga agataagcag 720
cagggacttg tacctgtctt cgtgtgcgcg actcttggaa ctacgggtgt ttgtgcattc 780
gatcgtctga gcgaactagg accaatctgc gcctccgagg ggttgtggct acacgtagat 840
gccgcctatg ctggcactgc tttcttatgc ccagagctga gagggtttct agagggtatc 900
gagtacgctg attcattcac ttttaatcca tccaagtgga tgatggtcca cttcgattgt 960
acaggcttct gggtaaagga caaatataag ttgcagcaaa cgttctctgt aaacccgatt 1020
tatcttaggc atgctaattc cggggcagcc accgacttca tgcactggca gatccccctt 1080
tctcgtaggt ttagaagcat aaagttatgg ttcgtaatca gatctttcgg tgtaaagaat 1140
ttgcaggcgc atgttaggca cggaactgaa atggccaaat atttcgaatc cttggttaga 1200
agcgacccca gtttcgagat ccccgcgaaa agacatctgg gtctagtagt tttcagactt 1260
aaaggtccta attgtttgac agagtcagtt ttgaaggaga tcgcaaaagc gggtcagtta 1320
tttttgatac cagcgacaat acaagacaaa ttgataatca ggtttacggt cacatcacag 1380
tttactacaa aggaggatat attacgtgat tggcacttaa tccaagaggc cgcaaatctg 1440
gtcctatctc agcactgcac ttctcagccc agcccccgtg cgaagaatgt gatcccaccc 1500
ccgcctggta cacgtggatt aagcttagaa tctgtttctg aaggaggtga tgatccggca 1560
caagcaagga aaataatcaa gcaaccaggg gctagcttgg cgcgtcgtga aggcggttcc 1620
gatttggaga caatgccgga ccccttcgac gattgtttca gtgaagaagc gccaaacacg 1680
acaaagcaca aattgtcaag ctttttattt agctatttat cagttcaaaa cagacgtaag 1740
acaacacgtt ccctgagctg taatagtgta ccaatgagcg cacaaaaatc attgccagct 1800
gacgcatctt tgaagaacgg tggaagtttc cgtgcaagaa ttttttctgg gtttcccgaa 1860
caaatgatga tgatgaagaa gggggcattc aagaagttaa taaagttcta tagtgtacct 1920
agtttcccgg agtgcagtag ccagtgcgcg cgtcaactac cctgttgtcc gttagaagct 1980
atggtt 1986
<210> 85
<211> 2541
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 85
atggacttta aggagtatag gcagagaggt aaagaaatgg tagactatat agcagactac 60
ttggaaaata ttcgtgaaag acgtgtattc cccgatgtct caccaggcta tatgcgtcag 120
ttattaccag aaagcgcccc catagagggg gagccttggc ccaaaatctt cagcgatgta 180
gaacgtatcg ttatgcccgg gataacccat tggcaatccc cgcatatgca tgcgtatttt 240
ccggccctga actcaatgcc aagtctgttg ggggatatgc tagccgacgc catcaattgt 300
ctgggtttta cctgggcttc ttcaccagcc tgcacggagc tggagatcat cgtgatgaac 360
tggttgggaa aaatgattgg actgcctgac gcttttctac acttatcctc ccaatcccaa 420
ggtgggggcg tcttacagac aactgccagc gaggccacgc tggtgtgcct tctagctggt 480
cgtactagag caatacagag gttccacgaa agacatcctg ggtaccagga cgctgaaata 540
aacgctagat tagttgccta ttgcagcgac caggcgcatt cttctgttga gaaagcggca 600
ctaataggcc tagtgagaat gagatatatt gaggccgatg atgatctggc gatgagaggg 660
aagcttctgc gtgaagctat tgaagacgac atcaaacaag ggttggtccc tttctgggtg 720
tgtgctacac taggcacaac cgggagctgt tcatttgata atctggagga gatcggaata 780
gtttgcgcag agcatcacct atggttacac gtagatgctg cttatgcagg atcagctttt 840
atatgtcctg agtttcgtac atggctgagg gggattgaac gtgcggacag catcgctttt 900
aacccatcca agtggttgat ggttcacttt gatgctacgg cactgtgggt cagggatagt 960
acagccgtac acagaacatt caatgtcgag cccctttatc tgcaacacga gaactctggt 1020
gtagcagttg attttatgca ttggcagatc ccattatcaa ggcgttttag agccctgaaa 1080
gtatggttcg ttctgaggag ctatggtata aaagggttac aaagacatat aagggaggga 1140
gtgaggttgg cacagaaatt tgaggctttg gtcttggccg atcacaggtt cgaactaccc 1200
gcgaaaagac acctgggact ggttgttttc cgtatcagag gcgataatga aataacagag 1260
aaattgctga agagattgaa tcatcgtggc aacttacact gtataccctc atccctaaaa 1320
ggacaatacg tcataaggtt tactatcacc tccactcaca cgacactgga tgatatagta 1380
aaagactgga tggaaatacg tcaggtcgcc agtaccgtgt tagaagagat gaacataacg 1440
attagcaata gagtttatct taaagagaca aaagaaaaga acgaggcgtt tggttcttct 1500
ttattgctgt ctaacagccc tctatcccct aaggtagtaa acggctcctt tgctgcaatc 1560
ttcgacgcgg acgagttcct agcgaaaaca tacgcaggcg tcaggatcgc tcatcaagaa 1620
tctcctagta tgcgtaggag ggtcagagga atcttgatgt ctggaaagca attttcctta 1680
gattcacaca tggatgttgt tgtccaaacc acacttgacg ctgggaatgg tgcaacgaga 1740
acttcaacca ctaatagcta cggacatacg acgagcgcgg cgcaggccaa ttccgaaagg 1800
caagctagca tacaagaaga taacgaagag tccccggaag aaactgaact tctgagtcta 1860
tgcaggacat ccaacgtccc gagtccagaa catgcgcatt cattgtctac accaagcagg 1920
tcctgttcat cttcatctca ctcccttatc cattcactta cccaatccag ccctcgtagt 1980
tcacccgtca atcagttcag gccgattact ctttgcgcgg tcccgtccca atctcaactt 2040
tccatgccgt tggctatgcc actaccaaat agaaacgtta cagtttctgt agattctctt 2100
ctaaaccctg taactacatg taatgtatat catggcaaaa gatttctgga gccactggaa 2160
aacctggccc agacgagcgc aagttttagc agctcaattt ttagacttcc aacaccgatc 2220
gcaacgccga ccagagagtc cccagaagat cctgattggc cggctaagac attcagtcaa 2280
ttgctgttgg agaggtactc cagccagtcc cagtcattag gtaacaactc tagcaccgaa 2340
tcctcatcat tgtccggagg ggctactccc acacctaccc ccatgtccag tttggatgaa 2400
ttggtcacac cgttattatt gtctttcgca tccccttctc aaccgatgct atctgcacat 2460
gggataggcg aaggtcaaag ggagcagggc tctgatagtg atgcaactgt gtgtagtact 2520
acaagctcta tggaaagcct a 2541
<210> 86
<211> 1161
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 86
atgtctcttt ccccacttga ccagaacagg attgaatctt tctggcaata ttgcttgcag 60
caccaatact tcaacttggc ctaccccgag tcagctgact tcgactatgc ccctttgcac 120
cgttttttgc gtttctccat taataactgc ggcgactgga atgagtcctc aaattacttg 180
ttaaattcat tcgatttcga gagggaagtt atgcacttct ttgccgaatt gttccacatt 240
ccgtttgacg agtcatgggg atacgtgacg aatggcggaa cggaagggaa tatgttcggg 300
tgctacctgg cgagggagct gtttcctgat gcaacccttt actatagtaa agactcccac 360
tacagtgtag ctaagattat aaagttgctg agaatcaaat ccagagcagt agactcacta 420
ccgagcggcg agatagatta cgacgacctt gtagccaaga ttcaacaaga ccaggagaga 480
catccaattg tattcgtaaa tgttggaacg actatgaaag gtgccgtaga tgacatagga 540
gttatccaac ataaacttgc ggaagctggt ataccgcgtc aagattacta cttgcacgcg 600
gatgcggctc tttcaggtat gatcttaccc ttcgtggacg caccacaacc gtactcattt 660
gcggatggga tagactcaat atctgtgagc ggtcataaaa tgataggaag ccctatgcca 720
tgtggtatag tattggcaaa gaggagcaac gtgagtcgta tatctgtaga aatcgattat 780
atatcagcga aagaccagac gatttctggg agtaggaacg gtcacacccc gatgatgttg 840
tgggctgcta ttaaatcaag gcccctagcg gaatggagaa gaaaggttag acattgctta 900
gacatggcac aatacgcaat cgacaggcta caggcggccg ggatacaagc ttggagatgt 960
aagaatagta ttaccgtagt tttccccagc ccctccgaac cggtctgcga caagcatggc 1020
ttagcccgtt ctgggggaac cgcacattta atcaccacgc cccaccacca cgatagtcaa 1080
agattggatc gtctgttaga tgacatagtg caagatttag gcgcgatgac tgcccctgct 1140
ggtgcgacaa tgtctgctgc c 1161
<210> 87
<211> 1002
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 87
atgcagaaat atgaatacga tccgaagtgg gtaattaata atgcaattag cagtgagcgt 60
gaattttgca ctggctacca aaatccaggt gcgtccggta atggatacgt caccacgatc 120
aaattgagca cagggctagt cgatattacc ccctgggaaa aagttcaagc gatgagtgaa 180
catgaggtaa taaaattcga tagagggtgc agtaatatcg tgagctacga taggtgtgaa 240
tgtaatgatg cctacattgg tgccattaat atgttgacgg ccagctcctt ttcaggactt 300
caaggagtta tctggggtta cgatatagct gttgtcgaga acctgcgttc cagaaaactt 360
tacgaccaaa agtggcccag tggtgatcct aatagtgaat acagtactcc agtctactca 420
atcgagccgc tgttgaacgc aactgagaga ctgtttggac acgcggagcc aggaaaaaga 480
aggtttaatc caatcccagg ttctatggta gtatgtgcca acaaatctgc cactagtgat 540
ccttcctcag acgtcaaaga aggctgggcg ttcagtgtca ttagcttagc aatcttggag 600
aatagaaatt ctggaagtaa cctattcatc gaagattgcg acattatcga tataaacaac 660
ccggacggaa cgcgtaagac gaaagaggac gtgaaggcta tgctggacac tactctacgt 720
aaggtgactg agtgcacggt gctatgtggg ttggaccagc acattaagta caaagaaata 780
ttcattggct acaaagtcat caagtttaat gagaagcaag ttgggtgcgc actagcatgc 840
gcgccttatg ttaccttggc gaggaacgct gttccccagg gtatgaagcc ttcaaagttg 900
acggatatga atatctctca gtgggagaat gcattaaatt tacaaccctt ggagaaaatt 960
gaaaaatcta agattggaat tctgggaatg ggcgttctgg at 1002
<210> 88
<211> 1047
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 88
atgaccctgc acagtagata tagcaagtta aagggtgaaa ataaaatgca aaagtacgag 60
tatgatccca agtgggttat caataacgct atttctagtg aaagagaatt ctgcacggga 120
taccagaacc ctggggcaag cggcaacggt tatgtgacaa ctatcaaatt gtctaccggc 180
ttagttgata taacaccctg ggagaaagtt caagcaatgt ccgaacacga ggttatcaag 240
tttgataggg ggtgttcaaa catagtcagc tatgaccgtt gtgaatgcaa cgatgcttat 300
atcggtgcaa tcaacatgct aacggcgtct agcttctctg ggctacaagg ggtcatatgg 360
gggtacgaca ttgcggtggt cgaaaatcta agatcaagaa agctgtatga ccagaaatgg 420
cccagtggcg atccaaattc cgaatacagc acgccagtgt actctattga gccacttctt 480
aatgccacag aaagactatt cggacacgca gaaccaggta aaaggagatt taatccaatc 540
ccaggatcaa tggtagtgtg tgctaacaag tccgcaacgt cagatccatc atcagatgta 600
aaagaaggtt gggccttctc tgtgataagc ttggctattt tggagaacag aaatagcggt 660
tctaaccttt ttatcgagga ttgcgacatt atcgacatta acaatccgga tgggacgcgt 720
aagaccaagg aggacgtcaa agcgatgttg gacactacat tgaggaaagt aacggaatgc 780
acagtcctgt gtggtttgga ccagcacata aagtataaag agattttcat agggtataag 840
gtaataaagt tcaatgagaa acaagttggt tgtgcactag catgcgcccc ctatgtaacc 900
ttagccagaa atgcagtacc acaagggatg aagccctcca agctgacgga tatgaacatt 960
agccagtggg agaatgcact taacctacaa ccgttggaaa agatagagaa aagcaaaatc 1020
ggcatcttgg gaatgggcgt gcttgat 1047
<210> 89
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 89
atgctaaaga tagcagtacc caacaaaggt agcctaagcg agagagctat ggaaatcttg 60
gctgaggctg gctatgctgg gagaggagac tccaaatctc taaatgtgtt tgacgaggct 120
aataatgttg agtttttttt tcttagacca aaggatatcg ccatatatgt cgcgggagga 180
cagctagacc taggcatcac cggcagggat ctagccaggg acagccaagc agatgtccac 240
gaggtcctgt ctttaggatt tggatcttct accttcaggt atgcagcccc tgccgatgaa 300
gagtggtcta tcgagaagct agatggcaaa cgtatcgcca caagttatcc caatttggtc 360
agggatgatc tagccgcgag aggtttatca gccgaggtcc tgagacttga tggggccgtc 420
gaggttagta tcaaactagg tgttgcggat gcgattgcag acgtggtatc caccggaagg 480
acgctacgtc aacaagggct tgccccattc ggtgaggtgc tgtgcacatc cgaggctgtt 540
atcgtgggaa gaaaagacga aaaggttaca cctgagcagc agatattact aagaaggata 600
cagggaattc tgcacgctca aaactttcta atgctggatt ataaggttga tagggataac 660
ctagacgccg ccacagccgt tactcctggt ttctccggtc ccgctgtcag tcctctagct 720
agggacaact gggtcgccgt cagggctatg gttccaagac gtagcgctaa cgcaataatg 780
gacaaattgg cgggccttgg ggccgaagca atactggcga gcgaaatcag gattgcacgt 840
att 843
<210> 90
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 90
atgttagata atactcgtct tagaatagcg atacagaaaa gtgggaggct ttccgatgac 60
tctcgtgaat tactagcgag gtgcgggatt aagataaacc ttcatacaca aaggttgatt 120
gcaatggccg agaacatgcc gatagacatt ttaagagtca gggatgatga tatcccagga 180
ctagtgatgg atggggtggt tgatttagga attattggag agaatgtttt agaggaagaa 240
ctgttgaacc gtcgtgcgca aggtgaagat cctaggtacc ttacccttag gagacttgat 300
tttggcggtt gccgtttaag cctagcaact ccagtcgatg aggcatggga cgggcccgct 360
gccctggatg gtaaacgtat agccacatcc tacccacacc tactgaaaag atatctggac 420
cagaaggggg tttctttcaa aagttgtttg cttaacggct ccgtcgaagt cgcgcctcgt 480
gcgggattgg ccgatgcgat atgcgatctg gtctccacgg gtgctaccct ggaggctaat 540
ggccttagag aagtggaagt aatatacagg agcaaagcgt gccttatcca gagagacggt 600
gaaatggccc agagcaaact tatagataag ctactaacga gaattcaagg tgttatccaa 660
gcaagagagt ccaagtatat aatgatgcac gcgccctctg agagattaga ggaggtcatc 720
gcattgttgc caggggcaga acgtcccacc atacttcctc tagcgggtga gcaacagagg 780
gtagccatgc acatggtgtc cagcgaaact ttgttctggg aaacgatgga aaagctaaag 840
gcccttggag cctcttcaat tctagttctg ccgatcgaaa agatgatgga g 891
<210> 91
<211> 738
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 91
atgaccttca ctattcttcc tgcagtcgat gtagttaacg gacaagcagt tcgcctagat 60
cagggcgagg ccggcactga aaaatcttat ggcacccctt tggaatctgc actgaagtgg 120
caggagcagg gtgcaaagtg gttgcacttt gtggacctgg acgcagcgtt caaccgtggt 180
tccaatcatg agatgatggc ggaaattgtc ggcaagctcg atgttgatgt ggagctcact 240
ggcggtatcc gtgatgatga gtctctggag cgcgcgctgg caaccggtgc acgtcgtgta 300
aacattggta ccgctgctct ggagaagcca gagtggattg cttctgcgat tcaacgctat 360
ggcgagaaga ttgctgtcga tatcgctgtg cgtttggaag atggtgaatg gcgcacccgt 420
ggaaacggtt gggtctccga tggtggcgat ctgtgggacg ttctcgagcg tttggattcc 480
caaggttgtg cacgtttcgt ggttaccgat gtgtccaagg acggcacctt gagtggtcca 540
aatgttgagc tgctgcgtga ggttgctgca gctacagacg cacctatcgt ggcatctggt 600
ggaatttctg ttttggaaga tgttttggaa ctagccaagt accaggatga gggcattgat 660
tccgtcatca ttggcaaggc actttatgag cacaagttca ccctcgaaga ggctttggct 720
gcagtagaaa agctcggt 738
<210> 92
<211> 606
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 92
atgactgtcg caccaagaat tggtaccgca acccgcacca ccagcgaatc cgacatcacc 60
gtcgagatca acctggacgg caccggcaaa gtagatatcg ataccggcct gccatttttc 120
gaccacatgc tcactgcatt cggcgtgcac ggcagttttg atctgaaagt ccatgccaag 180
ggcgacatcg agatcgacgc acaccacacc gtggaagata ccgccatcgt gctcggccaa 240
gcactccttg acgctattgg cgacaagaaa ggcatccgcc gtttcgcatc ctgccagctg 300
cccatggatg aggcattagt ggagtccgtg gtggatatct ccggtcgccc atacttcgtg 360
atctccggcg aaccagacca catgatcacc tccgtgatcg gtggacacta cgcaaccgtg 420
atcaacgagc acttctttga aaccctcgcg ctcaactccc gaatcaccct ccacgtgatc 480
tgccactacg gccgcgaccc tcaccacatc accgaagcag agtacaaggc tgttgcccgt 540
gcgctgcgcg gtgccgtaga gatggatcct cgtcaaacag gaatcccatc caccaaggga 600
gcgctc 606
<210> 93
<211> 1098
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 93
atgaccaaaa ttactttgag cgatttgcca ttgcgtgaag aactgcgcgg tgagcacgct 60
tacggcgcac cccagctcaa cgttgatatt cgcctcaaca ccaacgaaaa cccttaccca 120
ccgtcagagg cattggtcgc tgacttggtt gccaccgtgg ataagatcgc caccgagctg 180
aaccgctacc cagagcgcga tgctgtggaa ctgcgtgatg agttggctgc gtacatcacc 240
aagcaaaccg gcgtggctgt caccagggat aacctgtggg ctgccaatgg ttccaatgaa 300
attctgcagc agctgctgca ggcttttggt ggacctggac gcaccgcatt gggattccaa 360
cccagctatt ccatgcaccc aattttggct aaaggcaccc acactgaatt cattgcggtg 420
tcccgaggtg ctgatttccg catcgatatg gatgtggcgc tggaagaaat tcgtgcaaag 480
cagcctgaca ttgtttttgt caccaccccg aacaacccga ccggtgatgt gacctcgctg 540
gacgatattg agcgcatcat caacgttgcc ccaggcatcg tgatcgtgga tgaagcttat 600
gcggaattct ccccatcacc ttcagcaacc actcttctgg agaagtaccc aaccaagctg 660
gtggtgtccc gcaccatgag taaggctttt gatttcgcag gtggacgcct cggctacttc 720
gtggccaacc cagcgtttat cgacgccgtg atgctagtcc gccttccgta tcatctttca 780
gcgctaagcc aagcagccgc aatcgtagcg ctgcgtcact ccgctgacac gctgggaacc 840
gtcgaaaagc tctctgtaga gcgtgttcgc gtggcagcac gcttggagga actgggctac 900
gctgtggtgc caagtgagtc caactttgtg ttctttggag atttctccga tcagcacgct 960
gcatggcagg catttttgga taggggagtg ctcatccgcg atgtgggaat cgccgggcac 1020
ttgcgcacta ccattggtgt gcctgaggaa aatgatgcgt ttttggacgc agctgcagag 1080
atcatcaagc tgaacctg 1098
<210> 94
<211> 1326
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 94
atgttgaatg tcactgacct gcgaggtcaa acaccatcca agagcgacat ccgacgtgct 60
ttgccacgtg gtggcactga cgtgtggtct gtgcttccca tagtgcagcc tgttgtagaa 120
gatgtccaaa accgcggcgc tgaagctgct ttggattacg gcgagaagtt cgaccatatt 180
cgccccgcct cggtgcgggt gccagctgag gttattgctg cagcagaaaa caccttggat 240
ccgttggtgc gtgaatcgat tgaagagtcg attcgtcgcg tccgcaaggt tcacgctgag 300
caaaagccag ccgagcacac cactgaactt tcaccaggtg gcaccgtcac tgagcgtttc 360
atgccgattg atcgcgtggg actgtacgtt ccaggcggca atgcggtgta cccatcaagc 420
gtgattatga atactgtccc agctcaagag gctggtgtga actcccttgt ggttgcgtcg 480
cctcctcagg ctgagcacgg tggctggcct caccccacca ttttggcggc gtgttccatc 540
ttgggtgttg atgaggtgtg ggctgtcggc ggcggtcagg ccgtggcgtt gctggcttat 600
ggtgatgacg ctgcaggtct cgagcctgtg gatatgatca ctggacctgg caatatcttt 660
gtcaccgctg cgaagcgcct ggtcagggga gtggtaggta ctgattctga ggctggccct 720
acagaaatcg ctgtgcttgc tgatgcctct gccaacgccg tcaacgttgc ctacgatctg 780
atcagccaag cagaacacga tgtcatggct gcgtccgtgc tcatcactga ctccgagcag 840
cttgccaagg acgtaaacag ggaaatcgag gcgcgttact caatcacgcg caacgccgag 900
cgcgtcgcag aagctttgcg cggggcccag agtggcatcg tgcttgtcga cgacatttcc 960
gtgggtatcc aagtagccga tcaatacgca gcggaacacc tggaaatcca cactgagaac 1020
gcgcgcgccg tagcagagca gatcaccaac gcgggtgcga tcttcgtggg cgatttctca 1080
ccagtaccac tgggtgatta ctccgcagga tccaaccacg tgctgccaac ctctggatcc 1140
gctcgtttct ccgcaggtct atccacgcac atgttccttc gcccagtcaa cctcattgaa 1200
tacgatgagg ctgctctgaa ggacgtctcg caggttgtca tcaactttgc caacgccgaa 1260
gatcttccag cgcacggcga agcaatccgt gcacgctttg aaaacctccc caccaccgac 1320
gaggcc 1326
<210> 95
<211> 261
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 95
gtgaagacat ttgactcgct gtacgaagaa cttcttaacc gtgctcagac ccgccctgaa 60
gggtctggaa ccgtggccgc cttggataaa ggcatccatc atctaggtaa gaaggtcatc 120
gaagaagccg gagaggtctg gattgcagcc gagtatgaga ccgatgaaga gctagccgga 180
gaaatctccc agctcattta ttggacccag gtcatcatgg ttgctcgcgg cctgaagcca 240
gaagatatct acaagaacct g 261
<210> 96
<211> 774
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 96
atgggcgtgg caattcgagt tattccttgc ctggacgtgg acaacggccg ggttgttaaa 60
ggcgtgaact ttgaaaacct ccgcgatgct ggcgatcctg tggagttggc aaagcgctat 120
gacgaggaag gggcagatga gctgaccttc ttggatgtca ccgcctcgaa gcatggtcgc 180
ggcaccatgc tggatgttgt tcgacgcacc gctgatcagg tgttcatccc tctgactgtc 240
ggtggcggcg tgcgcagcga agaagatgtt gatcaattgc tgcgcgcggg cgccgacaag 300
gtttcggtga acacgtctgc gattgcccgt ccagaactgc tatcagagct gtccaagcgt 360
tttggtgctc agtgcatcgt gttgtctgtg gatgccaggc gcgttcctga aggtggaact 420
cctcagccat ctggttttga agtcaccact cacggcggtt ccaagtccgc agaacttgat 480
gcaatcgagt gggcaaagcg cggcgaagag ctgggcgttg gcgaaattct gctcaactcc 540
atggacggcg acggcaccaa aaacggcttt gacctagagc tgctggaaaa agttcgcgca 600
gccgtatcca ttcctgtaat cgcctccggc ggcgctggca aggcggagca tttcccacca 660
gctgttgcag ctggcgccaa cgcagtgctt gccgcgacca ttttccactt ccgcgaagta 720
accatcgccg aagtaaaggg agccattaaa gatgcaggat ttgaggtgcg gaaa 774
<210> 97
<211> 633
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 97
atgaccaaaa ctgtcgccct tctcgactac ggatctggaa accttcgttc tgctcaacgc 60
gcattagagc gtgctggtgc agaagttacc gtgagctctg atccagaagt ttgcaccaac 120
gctgatggcc tcctagttcc tggggtgggc gcatttgatg cctgcatgaa gggtttgaaa 180
aacgtcttcg gacatcgcat tatcggacag cgtcttgctg gtgggcgtcc agtgatgggt 240
atttgtgtgg gcatgcagat cctgtttgat gaaggcgatg agcatggcat taagtcagct 300
ggttgtggcg agtggcctgg caaggtggaa cgcctccagg cggagatcct gcctcacatg 360
gggtggaaca cacttgaaat gcctaccaac tcaccaatgt ttgagggaat ttcacctgat 420
gagcgtttct acttcgtgca ctcctatggt gtgcgcaagt ggacgttgga aaccgacgat 480
ctgaccacgc ctccagaggt tgtgtgggcg aagcacgaaa atgatcgttt tgtggctgct 540
gtggaaaacg gcacgctgtg ggctactcaa ttccacccag aaaaatcagg tgacgtaggc 600
gcaaagctac tgcgaaactg gatcaactac atc 633
<210> 98
<211> 354
<212> DNA
<213> 谷氨酸棒状杆菌（Corynebacterium glutamicum）
<400> 98
atgagtgaca atccacaaga gtatgagctg gattgggacg tcgaaaagcg attaaagctt 60
aacgacgccg gcctggtgcc ggcaatcgtc caggccgacg ggaccaacga ggtcctcatg 120
atggcctgga tggataccca cgcgctagcc tatactttgg cgacccgccg tggaacctat 180
ttttctaggt cccgcaacga gtactggatc aagggcctga cctctggaaa cgtccaagaa 240
gtcaccggac ttgccctcga ctgcgacggc gacactgtcc ttctgaccgt gaaacaaacc 300
ggcggtgcgt gccacactgg tgcccacaca tgtttcgaca atgacgtttt gctg 354
<210> 99
<211> 891
<212> DNA
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 99
atggatttgg tgaaccatct aaccgataga ctactgtttg caatcccaaa gaaaggtcgt 60
ttatattcta aaagtgtttc tattttgaat ggtgctgata ttacatttca ccgctctcaa 120
agattagaca ttgcactaag cacaagctta cctgtagcgt tgatctttct gcccgctgca 180
gatattccaa cttttgttgg tgaaggtaaa tgtgatcttg gtataactgg tgttgaccaa 240
gttcgtgaat ctgacgtcga cgtagactta gcaatcgatt tgcaatttgg taactgtaaa 300
ttgcaggtac aagtccccgt aaatggcgag tataaaaagc cagaacagtt aattggcaaa 360
accattgtta ccagtttcgt gaaacttgct gaaaaatact ttgccgattt ggaaggtact 420
actgttgaaa aaatgaccac aaggataaag tttgtcagtg gttccgtgga ggcatcatgt 480
gctctgggaa ttggtgatgc tattgtagat cttgtagaga gtggtgagac aatgagggca 540
gcaggtttag ttgatattgc caccgtccta agcacaagtg cctacctaat agaatcaaag 600
aacccaaaga gcgataagag tttgattgct actatcaaat caagaattga aggtgtcatg 660
accgctcaaa ggttcgtttc atgtatttat aacgcacctg aagacaagct gcctgaactg 720
ttgaaggtga cgcctggccg tagagcacca accatttcca aaattgacga tgaaggatgg 780
gttgctgtta gttccatgat tgagagaaaa acgaagggtg ttgttttaga tgaattgaaa 840
agactcggcg catctgatat catggttttc gaaatttcta attgtcgtgt a 891
<210> 100
<211> 1656
<212> DNA
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 100
atgccggtcg ttcacgtgat tgacgttgaa agtggtaacc tacagtcact aaccaatgca 60
attgagcatt taggttacga agtacaactg gtgaaatcac caaaggattt taacatatca 120
ggcacgtcaa gattgatttt gcctggtgtc ggaaattatg gccatttcgt cgacaattta 180
tttaatagag gattcgaaaa gccgataaga gaatacattg aatctggaaa accaataatg 240
ggaatttgcg tcgggctaca agcgctcttt gccggttccg tggaaagccc taagagtacg 300
ggtctgaact acattgattt taagttgtcc aggttcgatg attcagaaaa gccagtacca 360
gaaataggtt ggaattcttg cattccctcg gaaaacctat tctttggatt ggatccatac 420
aagaggtact atttcgtcca ttcttttgct gccattctga attcagaaaa gaaaaaaaac 480
ctagaaaatg acggttggaa aattgcaaaa gctaagtacg gttcagagga atttattgcg 540
gcagtcaaca agaataatat attcgctact cagttccatc ctgaaaaatc aggtaaagct 600
ggtttgaacg tcattgagaa ttttttgaag caacaaagtc ctccgattcc aaactatagt 660
gcggaagaga aggaactctt aatgaatgac tattcaaatt atggtctaac acgcagaatt 720
attgcttgtc ttgatgtacg tactaatgac caaggtgatt tggtggttac taaaggtgat 780
caatacgatg tacgtgaaaa aagtgatggt aaaggtgtta gaaaccttgg taagcctgtt 840
cagttggcac agaaatatta ccaacagggt gcggatgaag taacattttt gaatataact 900
tcttttagag attgtccttt gaaggatact ccgatgctag aggttctgaa acaagccgca 960
aagacagtct ttgttccatt gacagtcggt ggggggatca aggatattgt cgatgttgat 1020
ggaaccaaaa tacctgcttt agaagttgca agtctatact tcagatctgg tgctgataaa 1080
gtatcgatcg gtacggatgc agtctatgca gccgaaaaat actacgagtt gggtaacaga 1140
ggagatggaa cgtcaccaat agagacaatc tcgaaagcat acggtgctca ggcagttgtt 1200
atttctgtcg atcctaagag agtatatgta aattcacaag cagatacgaa gaacaaagtc 1260
ttcgagacag aatatccggg ccccaatgga gagaaatact gctggtacca atgtacaatc 1320
aaaggtggaa gagaatctag agaccttggt gtgtgggaat taacaagggc atgtgaagct 1380
ctaggtgctg gggagatttt attgaactgc atagacaagg atggctctaa ttctggttat 1440
gatctggaat tgatagaaca tgttaaagat gcggtcaaga ttcccgtcat tgcatccagt 1500
ggcgccggtg tacccgaaca tttcgaagag gccttcctaa agacccgcgc agatgcttgc 1560
ttgggtgcag gtatgttcca cagaggtgaa ttcactgtta acgatgtaaa ggagtattta 1620
ctagagcacg gattaaaggt tagaatggat gaagag 1656
<210> 101
<211> 1158
<212> DNA
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 101
atggtttttg atttgaaaag aattgttaga ccaaaaattt ataacttgga accttatcgc 60
tgtgcaagag atgatttcac cgagggtata ttgctagacg ccaatgaaaa tgcccatgga 120
cctactccag ttgaattgag caagaccaat ttacatcgtt acccggatcc tcaccaattg 180
gaattcaaga ccgcaatgac gaaatacagg aacaaaacaa gcagttatgc caatgaccca 240
gaggtaaaac ctttaactgc tgacaatctg tgcctaggtg tgggatctga tgagagtatt 300
gatgctatta ttagagcatg ctgtgttccc gggaaagaaa agattctggt tcttccacca 360
acatattcta tgtactctgt ttgtgcaaac attaatgata tagaagtcgt ccaatgtcct 420
ttaactgttt ccgacggttc ttttcaaatg gataccgaag ctgtattaac cattttgaaa 480
aacgactcgc taattaagtt gatgttcgtt acttcaccag gtaatccaac cggagccaaa 540
attaagacca gtttaatcga aaaggtctta cagaattggg acaatgggtt agtcgttgtt 600
gatgaagctt acgtagattt ttgtggtggc tctacagctc cactagtcac caagtatcct 660
aacttggtta ctttgcaaac tctatccaag tcattcggtt tagccgggat taggttgggt 720
atgacatatg caacagcaga gttggccaga attttaaatg caatgaaggc gccttataat 780
atttcctccc tagcctctga atatgcacta aaagctgttc aagacagtaa tctaaagaag 840
atggaagcca cttcgaaaat aatcaatgaa gagaaaatgc gcctcttaaa ggaattaact 900
gctttggatt acgttgatga ccaatatgtt ggtggattag atgctaattt tcttttaata 960
cggatcaacg ggggtgacaa tgtcttggca aagaagttat attaccaatt ggctactcaa 1020
tctggggttg tcgtcagatt tagaggtaac gaattaggct gttccggatg tttgagaatt 1080
accgttggaa cccatgagga gaacacacat ttgataaagt acttcaagga gacgttatat 1140
aagctggcca atgaataa 1158
<210> 102
<211> 660
<212> DNA
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 102
atgacagagc agaaagccct agtaaagcgt attacaaatg aaaccaagat tcagattgcg 60
atctctttaa agggtggtcc cctagcgata gagcactcga tcttcccaga aaaagaggca 120
gaagcagtag cagaacaggc cacacaatcg caagtgatta acgtccacac aggtataggg 180
tttctggacc atatgataca tgctctggcc aagcattccg gctggtcgct aatcgttgag 240
tgcattggtg acttacacat agacgaccat cacaccactg aagactgcgg gattgctctc 300
ggtcaagctt ttaaagaggc cctaggggcc gtgcgtggag taaaaaggtt tggatcagga 360
tttgcgcctt tggatgaggc actttccaga gcggtggtag atctttcgaa caggccgtac 420
gcagttgtcg aacttggttt gcaaagggag aaagtaggag atctctcttg cgagatgatc 480
ccgcattttc ttgaaagctt tgcagaggct agcagaatta ccctccacgt tgattgtctg 540
cgaggcaaga atgatcatca ccgtagtgag agtgcgttca aggctcttgc ggttgccata 600
agagaagcca cctcgcccaa tggtaccaac gatgttccct ccaccaaagg tgttcttatg 660
<210> 103
<211> 1005
<212> DNA
<213> 酿酒酵母（Saccharomyces cerevisiae）
<400> 103
atgcactcac accattcaca ctccggtgac tatagtgccc acggtacgga ccctttggat 60
tccgtggtcg atcaggtggt caacctcaac tttcacacgt actgtttgac agagcacata 120
ccaagaattg aggccaagtt tatatacccc gaagagcagt cattgggcaa gaatcctgag 180
gaagtcataa gcaagctaga aacatcgttc aagaatttca tgagtcatgc gcaagaaatc 240
aagactcgtt acgctgatag acccgatgtg cggactaaat tcattatagg aatggagatc 300
gaaagttgtg acatggctca tatcgaatat gcaaagcgac tcatgaagga gaataatgat 360
actttgaagt tttgtgtggg ttcggtccat cacgtcaacg ggatccctat tgatttcgac 420
caacaacaat ggtacaattc attgcattcc ttcaatgata atttgaaaga ttttctcctg 480
tcttacttcc aatcacagta cgaaatgctg atcaatatta aaccgttggt cgtgggtcac 540
ttcgaccttt acaaattatt tttgcccaat gacatgctag taaaccagaa atcgggcaac 600
tgcaacgaag aaaccggagt tcctgtagct tcactggacg tcatcagtga atggccagaa 660
atatacgatg cagttgtaag aaatttacaa tttatagact cctatggcgg cgcaattgaa 720
atcaatacgt ccgcattaag aaagggcctc gaggagccgt accccagcaa aaccttatgt 780
aatctggtca agaagcactg tggatccaga tttgttctaa gtgatgacgc acacggcgtg 840
gcgcaagtgg gtgtgtgcta tgacaaggta aagaaataca tagtagacgt gctacaatta 900
gagtacattt gctaccttga ggaaagccaa tcaccagaga atgtgttaac tgtaaagaga 960
ttacccattt cgcaattcgt taatgatccc ttttgggcca atata 1005
<210> 104
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 104
atggcaaaaa ttgataaaat tctcaatcag gaaggcattg accgcattgc aatcaatcca 60
taccagaaat acagccgtgg atatatggaa ccgggcaacc ttggaggagg ctacgtgacc 120
ggtctcaagg tggatgcagg cacccgtgaa aagactgacg actctatgct ggatggcatt 180
gtgtcttatg atcgcgccga atgcaaaaat gcctacatcg gccagatcaa catgatgacc 240
gcatccagct ttacgggcgt gcaaggccac atccttggat atgatctgct gcgcaaccct 300
gccgttgaca aggcccagcc cctcttttat gaaacacaat gggatggtag caaactgcca 360
atctatgacg gcaagcctct tcaagattcc ttggtagaat ttttcggtac cgcggacaat 420
cgacgccatt atccagcccc gggatctttt atcgtatgtg ctaataaggg tgttaccgct 480
gaacgaccac ttgaggatcg accattgaat cctggagaag catacggtgt gtggtcagca 540
atcgctattt caattgcaaa ggacccggtc cataactcct ctatgtttat cgaggatgcg 600
ggtacctgga acacgccgaa cgaagatgac cttaacgagt tcctgtatca tcgccgcgaa 660
gccattgcac gcagcattgc gcaatgtggc caagacgcct caacttcatt tgcatcctcc 720
tggattggct tcgcacatgt gatgatgaag ccaggcgaga ttggcaacgc cattacggtc 780
ggcccgtact tcagcatgcc tgttgacgcc gttccaggtg gctctattct cactccggat 840
gttgatatga acattatgga agacctctcc ctccctgaat ggcttgaaaa gatgggctac 900
caatccattg tggaaaacca agacatccag tat 933
<210> 105
<211> 1170
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 105
atgtgccgcc atgcgtgcat cgccttggca atgggaatga ttatgggcct ttcatcagag 60
gacgccggaa agatcgaatc gttctggcgc tattgcgtgc agcaccaata cttcaatatc 120
ggttatcccg aggcagctga ttttgactac tccgccctta atcgttttct gaacttctct 180
attaacaact gcggagattg gtcacagcaa tccaactacc tcctgaactc gttcgacttc 240
gaacgtgaag tcatgcagtt cttcgctact ctgttctgca tcccattcga acagtcctgg 300
ggttatgtga caaacggcgg cactgagggc aacatgttcg gttgctactt ggctcgtgag 360
ctcttcccag aagccaccct gtactactcc aaggacaccc attacagcgt cgctaaaatc 420
atccgactct tgcgcgtgaa atcttgcatg gtggactccc tcccgaatgg tgaaatgaac 480
tacgatgact tgatcaatcg catccgtctg gatggagaac gccacccgat catctttgca 540
aacatcggca ctaccatgac cggcgctacg gataacatcg caacgatcca acgccgtctc 600
aaaaagatcg gtattactaa gggcgattac tacctgcacg ctgacgcagc actgtctggc 660
atgatcttgc ctttcattga taacccacaa ccgttcagct tcgcagatgg tgttgactct 720
atttccgtgt ccggacacaa gatgattgga tctccgatcc cctgcggtat cgtgcttgca 780
cgccgtaaac acgtcgaaca cgtttctgtc gagattgatt acatttcagc ttgcgatcag 840
actatctccg gttcccgcaa cggttatact cctttgctgc tctggatggc cattaagtcc 900
cgctccttct ccgactggcg tcagcgtacc cagcactgcc tcgacatggc gcagtacgta 960
attgaacgct tccatgctaa gggtatccac gcttggcgca atcccaactc catcaccgtt 1020
gtcttcccga aaccagctga tcacatctgg aagaagcact gcctcgccac ctctggtaag 1080
atctcacaca tcatcacgat gccacatcac accggcaaag agactctgga tcgcgttatc 1140
aatgacatcg ctctcgaccg tgagccaaag 1170
<210> 106
<211> 1134
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 106
atgacgctga gcatcaacga ccaaaacaag ttggatgctt tctgggcata ctgtgttaaa 60
aaccaatatt ttaacatcgg ctaccctgaa agcgctgatt ttgactacac caaccttgag 120
cgctttttgc gcttttcaat caacaactgt ggtgattggg gcgaatattg caattacctg 180
ttgaactcct tcgacttcga gaaggaagtg atggaatatt tcgctgacct cttcaagatc 240
ccattcgaga agtcctgggg atacgtcacc aacggcggaa cggaaggcaa catgttcggt 300
tgctatctgg gccgagaaat ttttcccgat ggtaccctgt actactctaa agatacacac 360
tattcagtgg cgaagatcgt taagcttctc cgaatcaagt ctcaggtagt agaagcccaa 420
ccaaacggcg aaattgatta cgatgatctt atgaagaaga ttgcagcaga caaggaggcc 480
cacccaatta tctttgccaa tatcggtacc accgtacgcg gagctatcga cgatatcacc 540
gagatccaga aacgcatgaa ggccgccggc atcaagcgcg aggactacta cctgcatgca 600
gacgctgccc tctccggcat gatcctgcct tttgtggatg aacctcaggc cttcaccttt 660
gctgatggca ttgactccat tggcgtctcc ggtcataaga tgatcggcag cccgattccg 720
tgcggcatcg ttgtggcgaa aaaggaaaat gtggatcgca tctcagtgga aattgactat 780
atctcagcac atgacaaaac tatcactggc tcacgtaacg gccacacgcc actcatgctc 840
tgggaggcgg tgcgcgcgca ctctacggaa gattggaagc gtcgtatcgg tcgctctttg 900
gatatggcgc agtacgcagt agaccgtctg cagaaggccg gcatcaatgc ttggcgtaac 960
aaaaactcca ttaccgtcgt gtttccgtgc ccgagcgagc gtgtgtggaa ggaacactgc 1020
ctcgcaacct ctggtaatga tgcacatctg atcaccaccg cacaccattt ggacactgct 1080
cagattgatg ccctgatcga cgacgttatt gcagatgcca agctccacgc tgct 1134
<210> 107
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 107
atgcttaaga ttgcagtccc aaacaaaggt agcttgtccg agcgtgccat ggagatcttg 60
gccgaagcag gctatgccgg ccgtggcgat tcaaaatcgc ttaacgtctt cgacgaagcg 120
aataatgttg aatttttctt cctccgaccg aaagatatcg ctatttacgt ggcaggcggc 180
caattggacc tgggcattac aggacgagac ttggcacgag attcccaggc ggacgtgcac 240
gaggtactga gcctgggatt cggctctagc acctttcgct atgcagctcc agccgacgag 300
gaatggagca tcgaaaagct cgatggtaag cgcatcgcaa cctcttaccc aaacctggtg 360
cgcgatgatc tcgcggcccg cggcctttcg gcagaagtgc ttcgccttga tggtgcagtt 420
gaagtcagca tcaagctggg agtcgccgat gccattgcag atgtcgtctc tacgggacgc 480
acattgcgcc aacagggact cgctcctttc ggagaggtgc tctgcacctc ggaagcggtg 540
atcgtaggcc gcaaggacga aaaggtcaca ccagagcagc agatcctcct ccgccgaatt 600
caaggcatcc tgcacgcaca gaactttctt atgctggatt acaacgttga ccgcgataat 660
ctggacgcag cgaccgcggt cacccccgga ctctccggtc ccaccgtgtc gccacttgct 720
cgtgataatt gggttgcggt gcgtgctatg gtcccccgcc gcagcgcgaa cgctattatg 780
gataaactgg cgggacttgg cgcggaagcc atccttgcat ccgaaatccg cattgcgcga 840
att 843
<210> 108
<211> 945
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 108
atgcctgaca tgaaattgtt cgcgggcaat gcgacacctg agctcgctca acgaatcgcc 60
aatcgactgt acaccagcct tggagatgcg gcggttggac gcttttcgga tggcgaggtt 120
tccgtacaga tcaacgaaaa cgtgcgcggc ggcgacatct tcatcattca gtccacgtgc 180
gctcccacca acgacaatct gatggaactg gttgttatgg tcgatgccct gcgccgcgca 240
tcggctggtc gcatcacagc agtgattcca tattttggct acgctcgcca ggatcgacgt 300
gtgcgatcgg cccgcgttcc catcaccgcc aaagtcgtgg cggatttcct ttcctctgtg 360
ggagttgacc gtgtgctcac tgtagacttg catgcggaac agattcaagg cttctttgat 420
gtgcctgttg acaacgtttt cggcagccct atcctgctcg aagatatgct ccaactgaac 480
ctcgataatc ctattgtcgt ttcaccagat attggtggtg tcgttcgtgc tcgagcaatc 540
gcgaagcttc tcaacgatac cgatatggca attatcgaca aacgtcgccc acgcgcgaac 600
gtttcacaag ttatgcatat tattggtgat gttgccggcc gcgattgcgt gttggtggat 660
gatatgatcg acactggcgg cactttgtgc aaagcggcgg aagccctgaa ggaacgcggc 720
gccaaacgcg tatttgcata cgccacccac ccaattttct ctggcaatgc cgcgaacaac 780
ctgcgcaact ccgtgatcga tgaagttgtg gtgtgtgata cgatcccctt gtctgacgaa 840
atcaaatccc tcccgaacgt gcgcacactg actctctcag gcatgctggc cgaggccatt 900
cgacgaatct ctaacgagga gagcatctcc gcaatgtttg aacat 945
<210> 109
<211> 1488
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 109
atgtctatgt ccaatattgt cgttttcgga ggagactcgc acccagaatt ggtgaccaag 60
atttgcgaaa atcttgacat tcaccccagc aaggttgagc tgggcaaatt ttctaacggt 120
gaaactaaca tcgcactgcg tgaatcggtc cgagagaaag acgtctacat cattcagtcg 180
ggctgcggtc aagtgaacga tactttcatg caactgttga tcctcatctc cgcatgtaag 240
agcgcttccg catcccgtgt caccgcagtc atgccatacc tttgctattc gcgacaacca 300
gacatccctt acacagcgaa gggagcaccc attatttcca agccgaaaga aaactatacg 360
tttgaatcac atccaggaac ccccgtgtcc tcctccctta tgacccagcg acccggcgcg 420
gagtcgagcc ttaaaagctt ggattcagct atccgaagca ccatcaatct ggagaacccg 480
cagccaatcc gcactcccaa ttcttctgca accgcgaata acaactttga tattaagaaa 540
accctgagct tctcacgtat cccaatgatc cccggcggaa aactgcagaa tacttcgaac 600
tctaccgatg cgggcgaact cttcaatgct caaaacgccg gatacaagct ttgggttgtc 660
caagcaggaa ccctgatcgc acacctgctg tctgccgctg gtgccgatca cgtcatcacc 720
atggatctgc atgacccaca attcccagga ttctttgata ttcctgtaga taacctgtat 780
tgcaaaccca tcgctcaaaa ctatattcag caccgcattc ccgattacca agatgctgtt 840
atcgtgtctc ccgacgctgg cggcgccaag cgcgctaccg ctatcgcgga tgcattggaa 900
ctctcctttg ctttgatcca caaggaacgc cgaagccagc tgcttaaggg acctccagat 960
gccactctga cttcaggcgg ctctctgcct gtttcccccc gcccactcgt tacgaccctg 1020
gtctcctctc agaatacgac ttcctccggt gctacaggtg tggctgctct cgaaatgaag 1080
aaaactacct ccacaagctc cacctcatcg cagtccagca attcatctaa gtttgtccag 1140
accacaatgc ttgttggcga tgtccgcaat aaggtgtgca ttattgtcga tgacctggta 1200
gacactagct ataccattac ccgtgcagcc aaactcctca aggaccaggg ctcgacgaaa 1260
gtttacgccc tgattactca cggcgtattc agcggcgacg ctctggagcg cattggtcag 1320
agctccatcg ataagctgat catctctaac acagtccccc aggaccgtac cttgcaatac 1380
ctgggaaagg accgcgttga cgtaattgat gtgtcctgca tcatcggcga ggctatccgc 1440
cgtatccata acggagaatc gatctctatg ctgttcgagc atggttgg 1488
<210> 110
<211> 1281
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 110
atgcgtaaat gcaaaatctt cgtgggaaac tcacacccag aactgggaaa catggtatgc 60
caacgcttgg gtatcgagcc cgcaccttgt acccttaaaa agttcgccaa cggagagacc 120
tccgtccaaa tcggcgtgag cgtccgcgat gaagatgtct acgtgattca gtccggttcc 180
ccttcaatca acgatgacat tatggaattg ctgattctcg tttcagcgtg ccgaggcggt 240
tccgcccgta agatcaccgc tgtgatccca cagtttccat actcgaaaca gtgcaagatg 300
aaacgacacc gcggagcaat taccgcacgc atgctggcaa acttgcttgt catggcagga 360
gccgaccatg tggtatcgat ggatctgcat gcatcccaga tgcagggttt ctttaccaag 420
ccagttgaca acctttacgg aggcccgtca cttgccaaat ggatccgaga gaacgtcgaa 480
gactacgagg acgcagtagt tgtctcaaag aaccccggcg gaaccaagcg tgttaccgca 540
cttgcagact ctctcaagat caacttcgcc atgatccaca ccgatcgccg ccgtagcaag 600
gacctttatt cgcagaataa ggacctccaa caactgaagt tgcgtaaaca gtccatgctc 660
cgcaagaacc gtcccatcat tcgacagggc gatcatccaa acgaagaaga aaatattatt 720
ttgtcgaacg gcattcagac ggcccgtatc cgcaacggtc atgtgatcgg cgacgatgag 780
gccgacgatg acgaagatgc aattctcgaa agcgactcag agcttcactc catcgatggc 840
ctcgactcgc acggcctcgg aggcacttat gacgcagttg actccgaaga tgaagaagag 900
atcccggtgc tttaccgcga acagctcatt acattggtag gtaacgtgcg cggtcgcagc 960
gccatcatcc ttgatgatat gatcgatcgc cctggctctt ttatctccgc ggctgaacac 1020
ttggttcaga actgtggcgc aaagaaggtg tacgttgtgg ccacccacgg catttttacc 1080
ggtgattgtt tggaagagct cgagaagtcc gatgccatcg acactattgt ggttaccaac 1140
acctacccaa tttctggaga gcgcattgct ggtagcaaaa agctggttac catcgatgtt 1200
agccccatct tcgccgagtg cattcgtcgc gaccactatg gcgagtccat tagcgtgctt 1260
ttcgacagcc tggcggccct g 1281
<210> 111
<211> 2040
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 111
atgactcagt ttactgatat cgacaagctg gcagtttcta ccatccgcat cctggcggtc 60
gacaccgtct caaaggcaaa ctccggccac cctggtgcgc ctctgggtat ggctccagcg 120
gcccacgtcc tttggtcaca gatgcgaatg aatccaacga atcctgattg gatcaaccgc 180
gaccgcttcg ttctctccaa tggacacgca gttgctttgc tgtactcaat gcttcatctc 240
accggttacg atctgagcat tgaagatttg aagcaattcc gccagctggg ttcccgcacc 300
cccggtcacc ctgagttcga attgcccggc gttgaagtta cgaccggccc acttggacaa 360
ggtatttcca acgcagtcgg aatggctatg gcgcaggcga atctggcagc gacctacaac 420
aaaccgggtt ttacactctc cgacaattac acttacgttt tcctgggcga cggatgcctg 480
caagaaggca tctccagcga ggcgtcctcc ttggcgggcc acctcaaact gggtaacctg 540
attgcaatct atgacgataa caaaatcact attgacggcg ccacttccat ctctttcgac 600
gaggatgttg caaagcgcta cgaggcatac ggttgggaag tactgtacgt cgagaatgga 660
aacgaagatc tcgcaggcat cgcgaaagct atcgcccaag ccaagctgtc caaggataag 720
cccactctga ttaaaatgac gaccaccatt ggatacggct ccttgcacgc gggttcacac 780
tctgttcatg gtgctccttt gaaagccgat gatgttaagc aactcaaatc taagtttggc 840
ttcaatcctg ataagtcatt cgttgttcca caggaagtgt acgaccacta tcagaagacc 900
atcctcaaac ccggagttga agccaacaac aagtggaata agctcttcag cgaataccag 960
aagaagtttc ccgagctggg agcggaactt gcccgtcgtc tcagcggtca actgcctgcc 1020
aattgggaat ccaagctgcc cacgtacact gccaaggaca gcgccgttgc gacccgtaaa 1080
ctgtccgaaa ccgtcttgga ggacgtctac aaccaattgc ctgagctgat cggtggctcc 1140
gcggatctta caccgagcaa tttgacccgc tggaaggaag cgctcgattt tcagcctccc 1200
tcgtctggca gcggcaacta ctccggccgc tacatccgtt acggtatccg cgagcacgcc 1260
atgggcgcca tcatgaacgg aatcagcgca ttcggtgcca attacaagcc ttacggtggc 1320
acgttcctga atttcgtgag ctatgccgct ggtgcagttc gtttgtctgc actgtccggt 1380
cacccggtca tctgggtggc aactcacgac tctattggcg ttggagagga cggtccgacc 1440
catcaaccga tcgagactct ggcccacttc cgaagcctcc ccaacattca ggtctggcgc 1500
ccagccgacg gaaacgaggt ctctgcagcc tacaagaaca gccttgagtc caagcacacc 1560
cctagcatca tcgcgctctc tcgccagaat ctgccacagc tcgagggctc ctctatcgaa 1620
tccgcgtcca agggcggtta cgttttgcag gatgtggcaa atccggacat cattttggta 1680
gcaaccggct ccgaagtctc cctgtcagtg gaggccgcaa aaacactcgc ggcaaagaac 1740
atcaaggctc gagtggtctc gcttcccgat ttctttacct tcgataaaca gcccctggag 1800
taccgtttgt ctgtgttgcc agacaacgtg cccatcatgt cagtcgaggt actcgcaacc 1860
acgtgttggg gtaaatacgc gcaccagtcc ttcggcatcg atcgcttcgg agcgagcggc 1920
aaggcacccg aggtctttaa gttttttgga tttaccccag aaggagttgc agagcgtgcg 1980
caaaaaacca ttgcattcta taagggtgac aagctgatct cgccgctcaa gaaggcgttc 2040
<210> 112
<211> 1809
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 112
atgccgcgca acccgcttaa gaaggagtac tgggctgacg tggtcgatgg tttcaagcca 60
gccacctctc ctgcgttcga aaacgaaaag gaaagcacca ccttcgtgac agaactgact 120
tcaaagaccg actctgcgtt ccctctgtca tcaaaagata gccctggaat caaccaaacc 180
actaacgata tcaccagctc agaccgtttc cgccgaaacg aggataccga gcaggaggat 240
atcaataaca ccaacctctc gaaagatctg tcggttcgcc acctgttgac gttggcggtg 300
ggaggcgcta tcggcacagg tctctatgtg aataccggag ccgcgctttc taccggagga 360
ccagcttctt tggtcattga ctgggtcatc atttcaactt gtctgtttac tgtcatcaac 420
tccctgggtg agctgtcggc tgcgttccca gtggtgggag gatttaacgt ttactccatg 480
cgattcatcg agccctcatt tgcattcgcc gtgaatttga actacctggc gcagtggctg 540
gtgcttctgc cactggaact ggtggcagcg agcattacca tcaagtactg gaatgacaaa 600
atcaactctg atgcatgggt tgctatcttc tacgcaacca tcgcgctggc taacatgttg 660
gacgtgaagt catttggcga aaccgaattc gtgcttagca tgattaaaat tctgtcgatt 720
attggtttca ccatcctcgg catcgttctt tcatgcggag gtggccccca tggtggctac 780
attggcggca agtattggca tgaccccgga gctttcgtgg gccactcttc gggtacccag 840
ttcaagggcc tctgtagcgt ctttgtcacc gctgcattct cgtactctgg catcgaaatg 900
actgcggttt cggcagcaga aagcaaaaat cctcgcgaaa cgatcccaaa agcggcaaaa 960
cgcacattct ggttgattac cgcgagctac gttaccatct tgactctgat cggttgtctc 1020
gtaccaagca acgacccacg ccttcttaac ggtagctcat ccgttgacgc tgcatcctcc 1080
ccactggtaa tcgccattga aaacggcggc atcaagggct tgcccagcct tatgaatgct 1140
attatcctga tcgcggtcgt ctcggttgca aactcggcgg tgtacgcttg cagccgatgc 1200
atggttgcca tggcgcacat cggaaacttg cccaaattcc tgaatcgtgt ggataaacgc 1260
ggtcgtccta tgaatgccat tctgcttacc cttttcttcg gtctgctgtc ctttgtggcg 1320
gctagcgaca aacaagccga ggtgtttacc tggctttctg ccctgtccgg cctctccact 1380
atcttctgtt ggatggcaat caacttgtct catatccgct tccgtcaggc catgaaagtg 1440
caagagcgct ctctcgacga actgccgttc atctctcaga ccggcgtgaa aggctcctgg 1500
tacggattca tcgtgttgtt cctggtactt attgcatctt tttggacctc cctcttcccg 1560
ttgggaggca gcggcgcgtc ggcagagtcc ttcttcgaag gttacctttc cttccccatc 1620
cttatcgtct gctatgtggg ccacaaactt tacacccgta actggactct catggttaaa 1680
ttggaggata tggatttgga cactggccga aaacaagttg acttgactct ccgtcgcgag 1740
gaaatgcgta tcgagcgcga gaccctggca aagcgctctt ttgttacgcg atttctccac 1800
ttctggtgc 1809
<210> 113
<211> 2397
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 113
atggtgctcc caattctgcc tctgattgac gacttggcct cttggaactc taagaaggag 60
tacgtctctc ttgttggtca agttctcctg gacggatcca gcctgtccaa cgaggaaatc 120
ttgcaattct ctaaggaaga ggaggtgccg cttgtcgcgc tttccctgcc ttcgggtaag 180
ttctctgatg atgagatcat cgcgttcctc aacaacggag tatcctccct gttcatcgcc 240
tcccaggacg ctaagactgc tgagcatctt gtcgaacaac tgaacgttcc aaaggaacgt 300
gtggttgtag aagaaaacgg tgttttctcc aaccagttca tggtaaagca aaaattctcc 360
caagacaaaa ttgtttccat taagaagctg tccaaagata tgttgaccaa agaagtactg 420
ggcgaagtcc gtactgaccg cccagacgga ctctatacca ctctcgtcgt tgatcagtac 480
gaacgctgtt tgggtctggt gtattcctcc aagaaatcca tcgcaaaagc tatcgatttg 540
ggacgcggtg tctactattc gcgatcccgc aacgagattt ggattaaggg cgagacatcc 600
ggtaatggcc agaaacttct gcagatttca accgattgcg atagcgacgc tctcaaattt 660
attgtggagc aggaaaacgt cggcttttgc cacttggaaa ccatgtcatg tttcggtgaa 720
tttaagcacg gattggtcgg tctcgaatca ctcctgaagc aacgtctcca ggacgctcca 780
gaggaaagct atacacgacg cctgtttaat gactccgctc tgctggacgc aaagatcaaa 840
gaggaagccg aggagttgac agaagctaag ggaaagaaag aactctcttg ggaagcagcg 900
gacctttttt acttcgccct cgccaagctc gtcgccaacg acgtctcgct caaagatgtc 960
gaaaacaacc ttaacatgaa gcaccttaag gtgacccgtc gcaagggaga cgcaaaacca 1020
aaattcgtgg gccaaccgaa ggcagaggaa gaaaagctta ctggtcctat tcatctcgac 1080
gtggtgaagg cctccgacaa agttggtgtt cagaaggcac tgagccgccc aattcaaaaa 1140
acctccgaaa tcatgcacct cgttaaccca attatcgaaa atgttcgcga taaaggaaat 1200
agcgcactgc ttgaatatac cgagaagttc gatggtgtga agttgtcaaa ccctgtactg 1260
aacgccccat tcccagagga atacttcgaa ggacttactg aggagatgaa ggaagcactc 1320
gacctctcca tcgaaaacgt gcgcaagttc cacgccgccc agctgccgac ggaaacactg 1380
gaagtggaga cccagccggg cgttctgtgt tcgcgtttcc cacgcccgat tgagaaggtt 1440
ggcctttata tcccaggtgg cactgctatt ctgccctcaa ccgctttgat gttgggagtt 1500
ccggctcaag tggcgcagtg caaggagatc gtgttcgcct cccctccacg caagtccgat 1560
ggcaaggtct ccccagaagt ggtgtacgtt gctgaaaagg ttggcgcgag caaaatcgtg 1620
ctcgcgggcg gtgcacaagc agttgcagca atggcttacg gcaccgaaac catccccaag 1680
gttgataaga tccttggtcc aggtaatcaa ttcgtaaccg cggccaaaat gtacgttcag 1740
aacgataccc aggctctctg cagcatcgat atgcccgctg gtccgtcaga ggttctggtg 1800
atcgccgatg aagacgccga tgttgacttt gtggcctccg acctcctcag ccaggcggag 1860
cacggtattg atagccaagt aatcctcgtt ggtgttaacc tttcggagaa aaagattcaa 1920
gaaatccagg acgcggtgca caatcaagct ctgcagcttc cccgcgtgga tattgtgcgc 1980
aagtgcatcg cacactccac catcgttctg tgcgatggat atgaggaagc tttggagatg 2040
tcgaatcagt atgcaccaga acacctgatt ctccagattg caaacgctaa tgactacgtc 2100
aaactcgttg ataacgcagg ttctgtgttt gtaggtgcct acacccctga gtcctgcgga 2160
gattactcca gcggtacaaa ccacaccctg cccacctacg gttatgcgcg tcagtactct 2220
ggagcgaaca ctgccacctt tcaaaaattc atcacggctc aaaacatcac cccagaaggc 2280
ctggagaata tcggtcgagc tgtgatgtgc gttgcaaaga aagaaggcct cgatggtcac 2340
cgcaacgctg ttaaaatccg tatgtcaaag ctgggactga ttccgaagga cttccag 2397
<210> 114
<211> 1155
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 114
atggtgttcg acctcaaacg catcgttcgc cctaaaattt acaatctcga accatatcgc 60
tgtgctcgcg atgatttcac cgaaggcatc ctcctcgacg caaacgagaa cgcccacggt 120
cctacgcctg tagagttgtc taagaccaac ctgcatcgct acccggaccc acatcaactc 180
gagtttaaga ccgctatgac taagtaccgc aataagacca gcagctacgc gaacgacccc 240
gaggtgaagc cactcaccgc cgataacctg tgcctgggtg tgggttctga tgaatccatt 300
gatgcgatca tccgagcttg ctgcgtgcca ggtaaggaaa agatccttgt acttccgccc 360
acgtactcca tgtattcagt ctgtgcaaat atcaacgata ttgaagtcgt acagtgtccc 420
cttacggtct ctgatggctc tttccagatg gacaccgagg cggtcttgac catcctgaag 480
aacgattccc ttatcaagtt gatgtttgtg acctcaccag gaaacccaac tggtgccaaa 540
atcaaaacct ccctcatcga aaaggtcctc caaaactggg acaacggcct ggtcgtcgtc 600
gatgaagcct acgtggactt ctgtggtggc tcgaccgcgc cccttgttac gaagtacccc 660
aacctcgtga cactccagac cttgagcaag tcctttggat tggccggtat ccgccttggt 720
atgacttatg caactgccga gctggctcgc attttgaacg ctatgaaagc accgtacaac 780
attagctcct tggcgtcaga gtacgcattg aaggcggtgc aggactccaa cttgaagaaa 840
atggaagcta cgagcaagat tatcaacgaa gaaaaaatgc gtcttttgaa ggagctgaca 900
gcgcttgact atgtggatga tcagtatgtc ggcggtctgg atgcaaactt tttgctcatt 960
cgcatcaacg gcggtgacaa tgtcctggca aagaagctgt actaccaact cgcaacccaa 1020
tcaggcgtgg tggttcgttt tcgcggcaac gagttgggct gttcaggatg ccttcgcatt 1080
accgttggta cacacgagga aaatacacac ttgatcaagt actttaaaga aacgttgtac 1140
aagttggcaa acgag 1155
<210> 115
<211> 891
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 115
atggacctgg taaaccatct cactgatcgc cttcttttcg ccatcccaaa gaagggtcgc 60
ctgtactcta aatcggtatc catccttaac ggtgctgata tcacctttca ccgttcacag 120
cgcctggata ttgccctttc tacttctctc cccgtggcct tggtgttctt gccagcggcc 180
gacatcccaa cttttgttgg cgagggcaag tgtgatctgg gcattactgg cgttgaccag 240
gttcgtgaaa gcaacgtcga tgttgatctg gctattgacc tgcaattcgg caactgcaag 300
ctccaagtgc aggttcctgt taatggtgag tacaagaaac ccgaacagct cattggcaaa 360
accattgtta cctcctttgt taaactggct gagaagtatt tcgcggattt ggagggtacc 420
accgttgaaa aaatgacaac gcgcatcaag ttcgtgagcg gctcggttga ggcctcttgt 480
gctcttggca ttggagatgc tattgtcgac ttggtcgaat ccggcgaaac gatgcgtgca 540
gcaggtctgg ttgatatcgc gaccgtcctg tcaacctccg cctacctgat cgagtcgaaa 600
aaccccaaga gcgacaaatc ccttatcgca acgattaaat cccgtatcga aggagttatg 660
actgcacaac gcttcgtttc ttgtatttat aatgccccag aggacaaatt gcccgaactc 720
ctgaaggtga ctccaggtcg tcgcgccccc accatttcca agattgacga tgagggttgg 780
gttgccgttt cctctatgat cgagcgcaag accaagggcg tagtcctcga tgaactcaaa 840
cgactgggcg catctgatat catggtattc gaaatctcta actgccgagt g 891
<210> 116
<211> 783
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 116
atgacgaaat tcatcggatg catcgacctt cacaatggcg aggttaagca gattgtcggc 60
ggtacactga catccaagaa ggaagatgtc cctaagacta acttcgtttc ccaacatccg 120
tcttcttact atgcgaagct ttacaaagat cgcgatgtac agggctgtca cgttatcaag 180
cttggtccta acaatgatga tgctgcccgc gaagctcttc aggaatctcc ccagtttctt 240
caagtcggtg gtggaattaa cgataccaac tgcttggaat ggctcaagtg ggcttccaaa 300
gtcattgtaa cctcctggct ctttaccaag gagggacatt ttcaattgaa gcgcctggaa 360
cgtcttacgg agctttgtgg aaaggaccgc atcgttgtgg acctctcctg tcgtaaaact 420
caggacggcc gctggattgt agccatgaac aagtggcaga cccttaccga tctcgagctt 480
aacgccgata ctttccgcga actgcgcaaa tacactaacg aattcttgat tcacgcagca 540
gatgtcgaag gattgtgtgg cggaatcgac gaactgctcg tgtccaagtt gttcgagtgg 600
acaaaagact acgatgattt gaagattgtt tacgccggag gcgccaagtc tgtggacgat 660
cttaaacttg tggacgagct ctctcacggc aaagtggacc tcacgttcgg atcgtccctt 720
gatattttcg gtggcaacct ggttaaattt gaggactgct gccgttggaa cgagaagcaa 780
ggc 783
<210> 117
<211> 1656
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 117
atgcctgttg tgcacgtgat tgacgtggaa tctggcaatt tgcagtccct caccaatgca 60
atcgagcacc tcggttacga agtccagctt gtaaagtccc ccaaggattt taacatttcg 120
ggtacctccc gacttatctt gcccggcgtt ggtaattacg gacatttcgt cgacaatctt 180
tttaaccgag gttttgaaaa gccgattcgt gaatacatcg aatcgggcaa gcccattatg 240
ggcatttgtg tgggcctgca ggcccttttc gcgggttccg tcgaatcccc taagtccacc 300
ggccttaatt acattgattt caaactgtct cgcttcgacg attccgaaaa gcccgttccg 360
gaaatcggct ggaacagctg catccccagc gagaatctgt tcttcggact cgatccatac 420
aagcgatact actttgtaca ttctttcgct gcgattctga attccgaaaa gaaaaagaac 480
ctggaaaacg acggttggaa gatcgcgaag gccaaatatg gctctgaaga attcattgcc 540
gccgtgaata agaacaacat ttttgcaacg cagttccatc cagaaaaatc tggtaaagcc 600
ggtcttaatg tgatcgagaa cttcttgaaa caacagtcac ccccaatccc caactactcc 660
gctgaggaaa aggaactttt gatgaacgac tatagcaact acggactcac ccgacgtatc 720
attgcatgcc ttgacgttcg caccaacgac cagggtgatt tggttgtcac caaaggtgac 780
caatatgatg ttcgcgagaa gtccgacggc aaaggcgtac gcaatctcgg taagcctgtt 840
caactggctc agaagtatta ccaacaaggc gccgacgagg taacgtttct gaatattact 900
tcatttcgcg actgccctct gaaggatacc cccatgctcg aagttttgaa gcaggccgca 960
aaaactgtgt tcgtaccgct gaccgtggga ggcggcatca aagacatcgt agatgtggat 1020
ggtaccaaga tcccagcact cgaggttgct tccctttact ttcgatctgg tgcagacaag 1080
gtttcaattg gcacggacgc tgtctacgct gccgagaagt attatgagtt gggtaaccgc 1140
ggcgacggca catcgccaat tgagacgatc agcaaagcct acggtgcaca ggcagttgtc 1200
atctctgtag acccaaaacg cgtgtatgtc aattctcagg ctgacactaa gaacaaggtc 1260
tttgagactg aatacccagg tcccaacggt gaaaaatatt gctggtacca gtgcacgatc 1320
aaaggaggcc gcgagtcccg tgatcttggc gtgtgggaat tgactcgcgc atgtgaggca 1380
ttgggtgccg gcgaaatcct tcttaactgt atcgacaagg acggctcgaa ctccggttac 1440
gatctcgaac tcatcgaaca cgtgaaggat gccgttaaga ttcccgtgat tgccagctcc 1500
ggcgcaggag tgcctgaaca cttcgaggaa gccttcctga aaacccgcgc tgacgcatgc 1560
ttgggtgcgg gaatgtttca ccgtggcgaa ttcacagtca acgatgtaaa agaatatctt 1620
ctcgagcacg gccttaaggt tcgaatggac gaggaa 1656
<210> 118
<211> 744
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 118
atggctcgta ccttcttcgt gggaggtaac ttcaagctca atggttctaa acagtcaatc 60
aaagagatcg tggaacgact caatactgcg tccatcccgg aaaacgtcga ggtcgtaatt 120
tgcccacccg ccacctatct tgattactcc gtctcccttg tgaagaaacc acaagttact 180
gttggcgcgc aaaacgccta tttgaaggct tccggtgcgt tcaccggaga gaattcagtg 240
gaccagatta aagatgtggg cgcaaaatgg gttatcctcg gacactcaga gcgccgctcg 300
tacttccacg aggacgacaa gttcatcgca gacaaaacta agtttgcact gggacagggc 360
gtaggcgtca tcctttgcat tggagaaacc ctggaggaaa aaaaggcagg caaaacactc 420
gacgttgtcg aacgccaact caacgccgtt ctggaagaag tgaaggattg gactaatgtg 480
gtggtggcat atgaaccagt ctgggccgtg ggtaccggcc tggcggcgac ccccgaagat 540
gcgcaggaca ttcacgcgag catccgtaaa tttctggcct ctaagctggg tgataaggcc 600
gcttccgagc tccgcattct ttacggtggt tcagccaacg gctcgaacgc tgtgaccttt 660
aaggacaagg ccgacgtcga tggattcttg gtcggcggtg catctcttaa accagagttc 720
gttgacatca tcaacagccg caac 744
<210> 119
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 119
atgctgaaga tcgcagtgcc aaacaagggt tcgctttccg aacgagctat ggagatcctg 60
gcggaagctg gttacgccgg ccgtggagat tccaagagcc tgaatgtgtt cgatgaggcc 120
aacaacgttg aatttttctt tttgcgacca aaggacatcg cgatctatgt tgcaggtggt 180
caactggacc tgggtatcac cggtcgagat ctggcacgtg attcccaagc cgatgtgcac 240
gaagttctgt cgttgggctt cggctcttcc acgtttcgct atgccgctcc agctgatgaa 300
gaatggtcga tcgaaaaact cgatggaaag cgcatcgcga cctcatatcc aaaccttgtc 360
cgcgatgacc tggcagcacg cggattgtcg gccgaagtcc tccgtctgga tggcgcggta 420
gaagtcagca ttaagcttgg tgttgccgat gcaattgctg atgtggtgtc cacaggccgc 480
accctgcgcc aacagggcct cgcgcctttc ggagaagttc tgtgcacctc tgaagccgtg 540
atcgtgggcc gcaaagatga aaaggttact ccggaacaac agatcctttt gcgacgcatc 600
caaggcattc tccacgcaca gaacttcctc atgttggatt acaacgtgga ccgagataac 660
ctggatgctg caaccgcggt caccccaggc ctttctggtc ccaccgtctc cccgctcgcc 720
cgcgacaact gggtggcggt tcgtgcaatg gtaccgcgcc gatccgccaa cgcgatcatg 780
gataaactgg ccggcctcgg cgcggaggca attttggctt ctgaaattcg tatcgcacgc 840
atc 843
<210> 120
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 120
atgcttaaga ttgcagtgcc gaataaaggc tccctgtcgg agcgagctat ggaaattctg 60
gcggaagcgg gttacgcggg tcgcggtgat tctaagtccc tgaacgtctt cgatgaagcg 120
aacaacgttg aatttttttt tttgcgcccg aaggacattg caatctacgt agcgggaggt 180
cagctcgatc tgggtattac cggccgcgac ctggcacgtg attcacaggc tgatgtgcac 240
gaagtgctga gccttggctt cggctcctca acattccgct atgcggcacc agccgatgaa 300
gagtggtcga ttgaaaaact cgatggtaaa cgtatcgcaa cctcctatcc caacctggtc 360
cgtgatgatc ttgcggcgcg cggattgtcc gctgaagttc tccgactcga tggcgcggtc 420
gaggtatcca tcaagctggg cgtagcggat gcaatcgctg atgtcgtgag cactggccgt 480
accctccgcc agcaaggatt ggctccattc ggcgaggttc tctgtacttc ggaagctgtt 540
attgtgggcc gcaaggacga gaaagtgact ccagaacagc aaatcctcct gcgccgcatc 600
cagggtatcc tccatgccca aaactttttg atgcttgatt ataaggtgga ccgagataac 660
ctcgacgccg ccacggccgt gactccaggc ttttctggac ctgctgtttc gccgcttgca 720
cgtgataact gggttgcggt acgtgccatg gttcctcgtc gttctgccaa cgccattatg 780
gataagctcg caggcctcgg tgcggaagcg atcctcgctt ccgaaattcg cattgcgcgt 840
atc 843
<210> 121
<211> 948
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 121
atgtccgaat tcgacaagaa acttaacacc ctcggagtgg accgaatctc ggtatcaccc 60
tataagaaat ggtcccgtgg ttacctggaa ccgggtaacg ttggcaacgg ttacgtctcc 120
ggtctcaaag tcgacgcagg cgtgatcgat aaaacagatg acatgattct cgacggtatc 180
gtgtcgtatg atcgcgcgga gaccaaaaac gcctacatcg gccagattaa tatgactacc 240
gcgtcctcat tttctggagt gggcggcacc gttctgggct atgatatcct tcgaaatccc 300
gaggtcgata aagcgaaacc tctctttact gaaaaacagt gggatggttc cgagcttcct 360
atttatgacg cgaaaccact gcaggacacc ctggtggaat actttggcac caaagatgat 420
atgcgccatt accccgcacc gggcgcgttc gtttgctgcg caaataaagg agtgaccgcg 480
gaacgcccta aaaacgacgc cgatatgaaa ccgggacaag gctacggagt gtggagcgct 540
atcgctatca gcttcgcgaa agatcccaca aagtactcgt ccatgtacgt ggaagatgcc 600
ggtgtctggg aaacccctaa tgaggatgag ttgatcgaat acctgaaggg ccgacgcaat 660
gccatggcaa agtctatcgc cgcgtgcggt gagaacacag cagcagagaa cggcggagcc 720
gttttcacct caagctggat cggattcgct catgcaatga tgaaaccagg ccaggtcgga 780
aacgcgatta ccgtggcccc gtatatcgcc atgccagtgg actcaattcc aggcggttca 840
attcttacgc cagatactga catggatatt atgcagaatt tgacgatgcc tgagtggctc 900
gataagatgg gataccagcc cctcacgaag ggaggaaaca tcaactat 948
<210> 122
<211> 849
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 122
atggaaccag gcaacatcgg aaacggttat gtcaccggtt tgaaggtaga cgctggcgtt 60
cgcgacaaga ccgatgacga cgtgctggat ggaatcgtct catacgatcg tgctgaaact 120
aagaacgcat acatcggcca aatcaacatg accacggcaa gctccttcac gggcccgcag 180
ggccacgttg tgggatatga catcctgcga aatcctgagg tggacaaggt aaagccgctt 240
ttcgtggaaa agcaatggga tggctcggac ctgccaattt acgatgcaaa gccgcttcag 300
gacgccctgg tcgaatattt cggcatggag caggaacgcc gccactaccc agcaccgggc 360
tctttcatcg tttgcgccaa caaaggtgtc acggcggagc gccctaaagc tgacgagcca 420
cttaagccgg gtcagggcta cggcgtgtgg tccgccatcg ctatctcttt cgctaaggac 480
ccgtcgaaaa actcatccat gttcattgaa gacgcgggcg tgtgggaaac ccctaacgag 540
gatgagctca tcgagtatct gaatggccgc cgcaaggcaa tcgctaaatc tatcgctgag 600
tgcggtcagg acgctgatac cgctttcgaa tcttcctgga tcggctttgc ccatgttatg 660
atgaagcctg gtcagatcgg caatgcgatc accgttggac cgtacttttc gcttccggtg 720
gattccatcc ctaacggttc gattctgacc cctgataagg atatggagat catggagaac 780
ctttccctcc cggagtggct cgacaaaatg ggctatgagt cccttgtgaa gaagaataat 840
gtgacatac 849
<210> 123
<211> 1212
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 123
atgaccctct cgccagcgga tcagcgcaag cttgaaggtt tttggcagca ctgcgttacc 60
caccagtact ttaatattgg ataccccgaa tctgccgact tcgactattc ccagctgcac 120
cgattccttc agttctccat caataactgc ggagattgga atgaatactc taactacttg 180
ttgaactctt tcgacttcga gaaggacgtc atgacgtatt ttgcagaact cttcaatatc 240
gcactggagg actcctgggg ctacgtgaca aacggcggta ccgagggaaa catgttcggc 300
tgctacttgg gccgcgaact cttcccagat ggaacccttt actactcaaa ggacactcac 360
tactcggtgg ctaagattgt caagctcctc cgcatcaagt gtcgtgcagt tgaatctctc 420
ccaaacggag agatcgacta tgatgatctg atggctaaga tcaccgctga tcaggagcgc 480
cacccgatca ttttcgcgaa tattggtaca acgatgcgcg gcgctgtgga caatattgta 540
acgatccagc aacgccttca acaagctggc atcgcgcgcc acgattatta cctgcatgca 600
gatgctgccc tttccggcat gatcttgcct ttcgtggacc atccccagcc cttctcgttc 660
gctgacggca ttgactctat ctgtgtgtcc ggccacaaga tgatcggctc cccaattcca 720
tgcggcatcg tagttgcaaa acgcaataac gtcgctcgca ttagcgtcga agtcgactac 780
atccgtgcac acgacaagac aatttccgga tcgcgtaacg gccacactcc cctgatgatg 840
tgggcggcgc ttcgttctta ctcctgggca gaatggcgac atcgcattaa acactctctt 900
gatactgcgc agtacgcagt ggatcgattt caggcatcgg gtattgatgc gtggcgcaac 960
gaaaactcga tcaccgtcgt tttcccatgt ccatccgaac gtattgcgac caaatactgc 1020
ttggctactt ctggcaatag cgcccacctc atcacgaccc cgcaccatca cgactgttcc 1080
atgatcgacg cgcttatcga tgaagtcgtc gccgaggcac agctgaatac tctgcgctcc 1140
aaacgtgcgt tcacggacca gactgtggtt gagcgtctgc cagcggcaag ctttaacttg 1200
cgcacccatt ac 1212
<210> 124
<211> 1224
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 124
atgactttgt ctacggcaga cactcatcgt cttgatgact tttggcagca ctgtctcaag 60
caccagtttt tcaacattgg atatcctgaa aacgccgatt ttgactattc cgcgctggag 120
cgtttcctgc gattctccat caataactgc ggagactggt cggagcactc caactacgta 180
ttgaactcct ttgatttcga gaaagaagtc atggcttact ttgcagactt gtttgagatt 240
ccacgtgaag actcatgggg ctacgtgact aacggcggaa ccgaaggcaa catgttcggt 300
tgttatctcg ctcgcgaact gttccctgat ggtaccctct actactcaaa ggatacccac 360
tactctgtag ctaagatcgt aaagcttctc cgcatcaagt gccgcgctgt gaacgctctg 420
ccaaccggcg aaatcgatta cgacgacctg ctcgccaaga tcgcggccga cggtgagcgc 480
catccaatta tctttgcaaa cattggcact actatgcgcg gcgcagttga cgatatcgcc 540
gttattcaac agcgcttgca ggacgcaggc attgcgcgcc gcgattatta cttgcacgct 600
gatgcggcac tgtccggtat gattctccca ttcgtcgacg caccacagcc tttcactttc 660
gccgacggta tcgattccat ctgcgtctcc ggccacaaga tgatcggttc cccgatccca 720
tgtggtatcg ttgtggctaa gcgtcgcaat gtagcccgaa tctccgtgga ggtggactat 780
atctccgcct ccgataaaac tatctccggt tcgcgcaacg gccatacccc catgattatg 840
tgggcagcat tgcgtagcca ctcgtccgcg cagtggcgac gccgcgtcga gcgttccctg 900
gcggccgcac agtacgcggt gaaccgcctg caggccggcg gtgtcaaggc ttggcgtaac 960
cctaacagca tcacggtggt cttcccctgt ccgtcggcag atatcgctcg aaagtacggc 1020
ttggctactt ccggtgacac cgctcatctc attaccaccc cacatcatcg cgataatcgc 1080
gctattgacg ctctgatcga tgaagttatc gcggatgcgc gccccgaggt ctggcgagca 1140
acattgcaac gctcgtggca gggttccgct cagcgcttgc cacgaaccgc atcctggaac 1200
caggcggccg gactgggccg cttt 1224
<210> 125
<211> 1281
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 125
atgacatttt cgccagcaga ccacaaacgc cttgaggctt tctggcagta ttgtcttacc 60
caccagtatt tcaacgtggg ctatccggaa tccgcagact tcgactactc actccttcac 120
cgcttcatgc gtttttccat caataactgc ggcgattgga acgaacccag caactacttg 180
cttaacagct tcgattttga acgcgaagtt atgcgatttt ttgcggaact ctttcacatc 240
ccctttgaag actcgtgggg ctacgtgacc aacggaggaa ccgagggaaa tatgttcgga 300
tgttaccttg cgcgcgaact ttttccggac gcaaccctct actactccaa agatacgcac 360
tactccgtcg ctaagatcgt ccgtctcctg cgcatcaagg cacaggtcgt ggaatcacaa 420
gcaaacggag agatggacta tgatgatctt gtggcccgta ttgcagccga cggcgaacgt 480
caccccatca tcttcgccaa tattggtacc acgctgcgcg gtgccaccga taatattgcg 540
gtcatccaac aacgtctggc tcaagccggc atccgtcgcg aggattacta cttgcacgcc 600
gatgccgcac tgtcgggcat gatcttgcca tttgtggatg ctccagaacc atattcattt 660
gcggatggca ttgactccat ctgtgtgtcg ggacacaaga tgattggctc tccaatgcct 720
tgcggcatcg tcgttgctcg tcgccacaat gtagaacgaa tctccgtgga aatcgactac 780
atctcagcac gagatcagac catttcaggt agccgcaacg gtcacactcc acttatgatg 840
tgggctgcct tgtgctcccg ctctcgcgag gactggcgcg cccgtatcca gcgttgtctc 900
gatttggcgc aacacgcggt ggatcgcctg cgtgcagctg gcatcgaggc atggcgcaac 960
ccaaactcca tcaccgtcgt ttttccatgc ccctctgcct cggtctggaa gcgccattgc 1020
ctcgcaacca gcggcgatac cgcacacttg atcactactg ctcatcatca ggattccacc 1080
cagatcgatg cactgttgga tgagttgatt gccgacttga aggcgcgcac ccgctaccct 1140
ttgcaggccc ccctggtcga aggtctgggt tcacgaacgc gcgtgtattc cactgcaggc 1200
cagccgcgcc cggagccact ccctgatcac gcacgccccg cgcgcgattc aggacttgca 1260
cgtctgaaag actccgaaat c 1281
<210> 126
<211> 1161
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 126
atgtcgctga gcccgcttga ccaaaaccgc attgaaagct tctggcagta ttgccttcag 60
catcagtatt tcaaccttgc ctatcctgaa tccgccgact ttgactatgc cccccttcac 120
cgtttcttgc gattctctat caataactgc ggtgattgga acgagtcctc caactacctc 180
ctcaattcgt tcgacttcga gcgcgaagtg atgcacttct tcgcggaatt gtttcacatt 240
ccatttgatg agagctgggg ttatgtcacc aacggcggaa ccgagggcaa catgttcggt 300
tgttacctgg cccgcgaatt gtttccagat gcaacattgt actattcgaa agattcacac 360
tactcggtcg ctaagatcat taagctgctg cgtattaagt ctcgtgccgt ggactccctc 420
ccatccggtg agatcgatta cgacgatctt gtcgcaaaaa tccaacagga tcaggaacgc 480
catcccattg tttttgttaa tgtaggtact accatgaagg gagcagtcga tgacatcggc 540
gtgatccagc ataaactcgc tgaagcggga attccccgtc aggactacta cctccacgcg 600
gacgcagcat tgtcgggtat gatcttgccc ttcgttgacg cgcctcagcc gtactcattc 660
gcagacggaa ttgattccat tagcgtgtcc ggccacaaga tgatcggctc tccaatgccc 720
tgcggcatcg tactggcaaa gcgctcgaat gtctctcgca tttctgttga gattgattat 780
atctcggcca aggatcaaac catctccggc tctcgtaacg gccacactcc aatgatgttg 840
tgggcggcaa tcaagtcccg tccgctcgcg gaatggcgcc gcaaggtccg ccactgtctc 900
gacatggcgc agtacgcgat tgatcgactc caggcggcag gtatccaggc atggcgctgc 960
aagaactcga ttactgtcgt tttcccatct ccttcagaac ccgtttgtga taaacacgga 1020
cttgctcgca gcggcggtac ggcgcacctc atcacgaccc cacatcacca tgatagccaa 1080
cgccttgacc gccttctgga tgacatcgtt caggatctcg gtgcaatgac tgctccggct 1140
ggagctacaa tgtctgccgc g 1161
<210> 127
<211> 1134
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 127
atgaccttgt ccattgttga tcagaataag ctggacgctt tctggtctta ctgtgttaag 60
aaccaatact tcaacatcgg ttatccggaa agcgccgatt ttgattatac catcctcgaa 120
cgtttcatgc gcttcagcat caacaactgc ggtgattggg gcgagtattg taactacctg 180
ctgaactcct tcgacttcga aaaagaagta atggaatatt tcgcccgtat ctttaagatt 240
cctttcgaag agtcgtgggg ctatgttacc aatggtggca ccgaaggcaa catgtttggc 300
tgttacttgg gccgtgaatt gttccctgaa ggaactcttt actattccaa ggacacgcac 360
tatagcgttg cgaaaattgt gaagttgctt cgcattaaat cctccctcgt agaatcgcaa 420
ccaaatggag aaatggatta cgacgatttg atccgcaaaa ttcaacgcga taatgaagaa 480
catcccatca tcttcgctaa cattggcacc accgttcgtg gagctatcga caacattgca 540
gaaattcaac aacgcattgg acagttggga atcaaacgag acgactacta tttgcacgcc 600
gatgctgccc tgagcggaat gattttgcca tttgtcaacg acccacaacc atttaatttt 660
gcagacggca tcgacagcat tggtgtgtca ggacacaaga tgatcggttc ccctattcca 720
tgtggcatcg tcgttgctaa acgtaagaac gtggatcgaa tctccgtcga gatcgattac 780
atttccgcgc atgataaaac catctccggc tctcgcaatg gtcatacacc actgatgatg 840
tgggaagcta tccgctcgca ctcttggagc gattggcagc gccgcatcga gcactcgctc 900
aacatggccc aatacgccgt ggatcgcctc caagctgctg gtatcgacgc atggcgtaac 960
aagaatagca tcaccgtcgt gtttccatgt ccgtccgaag cagtgtggaa gaagcactgc 1020
ttggctacga gcggcgatat cgctcatctt atcacaactg cacatcacct cgacagctct 1080
aagattgatg agttgattga cgacgttatt gctgatttga accagcaagc cgca 1134
<210> 128
<211> 1161
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 128
atgagcctgt cctccttgga tcaaaaccgc attgaatctt tctggcaata ttgtctccag 60
caccagtatt ttaaccttgc ctatcccgaa tccgccgact tcgattacac cccgttgcat 120
cgctttttgc gattctcaat caacaattgt ggtgattgga acgaaagctc caactacctc 180
cttaactctt tcgatttcga gcgcgaagtc atgcacttct tcgcggaact cttccacatc 240
cccttcgacg agtcctgggg ctacgtgacg aacggtggca cggagggaaa catgtttggc 300
tgctaccttg cccgagagtt gttccccgat gctaccctct attacagcaa ggatagccac 360
tactctgtgg caaaaatcat caaactcctg cgcattaaga gccgcgcggt ggactctctt 420
ccctccggtg agatcgatta tgacgatctt gtcgccaaga tccagcaaga ccaagagcgt 480
catccaatcg ttttcgtcaa tgtgggcacg acaatgaagg gtgctgtgga cgacattggt 540
atcattcagg ataaactggc acaggcgggc attccgcgtc gcgattacta cctgcatgct 600
gacgcggcac tcagcggtat gatcctccca tttgtagacg cacctcaacc ttattctttt 660
gcagatggca tcgactccat ctccgtcagc ggtcacaaga tgatcggctc ccccatgcca 720
tgtggcattg tcttggcaaa acgttccaat gtatcgcgca tctccgtgga gatcgattac 780
atctctgccc gcgatcagac aatctctggc tcccgtaacg gacacacccc gatgatgctg 840
tgggcagcaa tcaaatctcg ccccttggca gaatggcgcc gtaaagtacg ccactgtctg 900
gatatggctc agtacgccat cgaccgcttc cgcgccgcag gcattcaggc ttggcgctgt 960
caaaattcaa tcaccgtcgt cttcccatcg ccctccgagc ccgtctgcga taagcacggt 1020
ctcgcgcgct ccggcggtgc cgcccacttg atcacaactc cacatcacca tgattcccag 1080
cgcctcgacc gtctcatcga cgatatcatt caggatctgg gtgcagtaac ggctcctgcc 1140
ggtgccgcta tgtccgcagc a 1161
<210> 129
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 129
atgtctgaat tggatgccaa actgaacaaa ttgggagtcg atcgcattgc gatttcccca 60
tacaaacagt ggactcgagg atatatggaa ccaggcaaca ttggtaacgg ttacgtcacc 120
ggcttgaagg ttgatgcagg cgttcgcgat aagagcgatg atgatgtgct cgacggtatc 180
gtgagctacg atcgtgcaga aaccaagaac gcctatatcg gacagattaa tatgactacc 240
gcgtcatcat tcactggtgt tcagggtcgc gtgatcggtt acgatattct gcgatccccg 300
gaagttgaca aggcaaagcc cttgttcaca gaaacccagt gggacggttc cgaactgccc 360
atctacgacg ccaaaccttt gcaagacgct cttgtcgagt attttggcac cgaacaggat 420
cgtcgacact accctgctcc gggttctttc attgtctgcg ctaataaagg tgtgaccgct 480
gagcgtccca aaaacgatgc ggatatgaaa ccaggtcaag gctatggtgt ctggtcggcc 540
attgcgattt cgtttgccaa ggacccaacg aaggactcgt ccatgttcgt tgaagatgca 600
ggtgtttggg aaacccccaa cgaggatgag ctgctcgagt accttgaagg ccgtcgaaaa 660
gctatggcga aatctatcgc ggaatgcgga caggatgctc acgcctcatt cgaatcttcg 720
tggatcggtt ttgcgtacac aatgatggaa ccaggtcaga tcggaaatgc tattacagtt 780
gcgccctacg tgtccttgcc tattgattca attcctggtg gttctatcct cactccggat 840
aaggatatgg agattatgga gaatctcacc atgcctgaat ggcttgagaa gatgggctac 900
aaatccttga gcgctaacaa tgctctcaaa tat 933
<210> 130
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 130
atgtccgagc ttgacactaa attgcacaag ttgggcgttg atcgcattgc aatttctcct 60
tataagcaat ggtcccgagg atacatggag cccggcaata ttggtaacgg ttacgtaacc 120
ggtttgaagg tggacgccgg cgtgcgcgac aagaccgacg atgaagttct cgacggaatt 180
gtgtcatacg atcgagcaga aacgaaaaac gcatacattg gccagatcaa catgacgacc 240
gcctcgtcct tcaccggccc acaaggacac tgcattggct acgatcttct ccgcaaccct 300
gaggtggaca ccgccgaacc tctcttcact gttaagcaat gggatggcag cgaattgccg 360
atttacgacg ccaaaccgct gcaggattct cttgtggagt atttcggcac caacaacaac 420
cgccgccact acccagcgcc tggctcgttc attgtttgcg caaacaaggg tgtaactgcc 480
gaacgcccaa tgaatgattc cgatatgaaa ccgggccagg gctacggagt gtggagcgca 540
attgcgcttt cctttgcgaa ggatccggca aaagactcct caatgtttat cgaagatgcg 600
ggcgtgtggg aaactccgaa tgaggacgag cttatcgagt atcttaaggg acgtcgtaag 660
gcgattgcaa aatccattgc agaatgcggt caagatgcca acacatcctt caaaggctct 720
tggatcggct ttgcccacgc catgatggaa ccgggccaga tcggaaacgc aattacagtt 780
gctccttata tttctatgcc ggtcgatagc attccaggcg gctcaatcct gactccggat 840
accgatatgg acattatgga aaatctcacc atgccagaat ggttggataa aatggagtac 900
aaatccctga ctgctaacgg tgcaatcaag tac 933
<210> 131
<211> 954
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 131
atgtccacta actcgatcaa actgctggct ggaaactcgc acccaggcct tgccgaactg 60
atctcgcagc gacttggcgt gccgctcagc aaagtcggcg tgtaccagta ctctaacaag 120
gagacctcag tcacaattgg cgaatccatt cgcgacgagg acgtttacat tatccaaacc 180
ggttacggtg agcatgaaat caacgatttt cttatggagc tcctgatcct gattcacgct 240
tgcaagacag ccagcgtgcg ccgtatcacc gcggtcatcc ctaactttcc gtacgcacgc 300
caagataaga aagataagtc ccgtgcgccg atcaccgcca aattgatcgc caacctcctg 360
gagacggccg gctgcgatca cgtgatcacc atggacttgc atgcatcgca gattcagggt 420
ttcttccaca tcccggttga caacctttac ggcgaaccgt cagtgctgaa ctacatccgc 480
acgaagaccg attttaataa cgcaattttg gtatcgcccg acgcaggcgg cgcgaaacgt 540
gttgcgtctt tggcagacaa gctggatatg aactttgctt tgatccacaa ggagcgccag 600
aaggcaaacg aggtctcccg catgctgctc gtcggtgatg tcgcgggcaa aagctgcctt 660
cttatcgacg atatggcaga cacatgtggt actctggtga aggcttgtga caccctgatg 720
gaccatggcg ccaaggaagt tattgcaatc gtgacccacg gcattttctc gggttcagct 780
cgcgagaaac tcattaattc gcgcctgtcc cgcattgtct gcacgaatac cgtaccggtg 840
gatctcgatc tcgacatcgt ggatcaggtc gatatctcac caacgattgc tgaagcaatt 900
cgccgtctcc acaacggcga atccgtttcg tacctcttca cgcatgcgcc agtc 954
<210> 132
<211> 960
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 132
atgcctacaa attcgatcaa gttgttggcg cccgatgttc accgcggtct tgcggagctg 60
gttgctaaac gcttgggact gcaacttact tcctcaaagc tgaagcgaga ccctaccggc 120
gaggtgtcct tcagcatcgg cgaatcagta cgcgaccagg atatcttcat catcactcaa 180
attggctccg gcgtggtgaa cgatcgtgtc ctggaactgc tgatcatgat caacgcctcg 240
aaaaccgcgt ccgcccgacg catcaccgcg atcatcccca attttccgta tgcgcgccag 300
gaccgcaagg ataagtcccg tgctccaatt accgctaaac ttatggcaga catgctgacc 360
accgcaggct gtgaccatgt tatcactatg gatctccacg cgtctcagat ccagggtttt 420
ttcgacgtgc cagtggataa tctgtacgcg gaaccatccg tcgtgcgcta tatcaaggaa 480
aacgtgaatt atatggattc cattatcatc agccccgatg cgggaggcgc caagcgcgcg 540
gcaaccctcg cggatcgcct ggatttgaat ttcgcactca tccacaagga gcgcgcgcgc 600
gctaatgaag tctcccgcat ggtgctcgtt ggcgatgtta ctgataagat ctgcattatc 660
gtagatgaca tggctgacac ttgtggaact ttggccaagg cagccgagat tctgctggaa 720
aaccgcgcga agtctgttat cgcgattgtc acccacggcg tcctttcggg tcgcgcaatt 780
gaaaatatca ataactccaa actcgatcgc gttgtgtgca ccaacacagt tccatttgaa 840
gaaaagatca aaaaatgccc caagctggct gtcatcgata ttagctctgt actggcagag 900
tccatccgcc gccttcacaa cggtgaatca atctcctatc tcttcaagaa ctatccgttg 960
<210> 133
<211> 1005
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 133
atgtccgaac cagcccaaaa aaagcaaaag gttgctaaca attccctgga gcaactgaag 60
gcctctggta ctgttgtagt ggctgatact ggtgatttcg gttccatcgc caagtttcag 120
cctcaggatt ccactacaaa cccctccttg atccttgccg ccgctaagca accaacctac 180
gccaagctca tcgacgtcgc agttgaatac ggcaagaaac acggtaagac tacggaggag 240
caagtcgaga acgcggtcga ccgcctgttg gtggagttcg gcaaggagat ccttaaaatc 300
gttcccggcc gcgtctccac cgaagtcgat gcccgattgt cctttgatac ccaagccacg 360
atcgaaaagg cccgtcacat cattaagctc tttgagcagg aaggcgtttc caaagaacga 420
gtcctcatta agattgcgtc aacctgggaa ggcatccaag cagccaaaga gcttgaggag 480
aaggacggta ttcattgtaa tctgacgctg ctcttttctt tcgttcaggc tgtagcttgt 540
gcagaggcgc aggttactct catctcccca ttcgtaggtc gaatcctgga ttggtacaaa 600
tcttcaaccg gcaaggatta caagggcgag gctgaccctg gcgtcatctc cgtcaaaaag 660
atctataact actacaagaa gtatggctat aagaccatcg tcatgggcgc ctctttccgt 720
tccaccgatg aaattaaaaa ccttgctggc gttgactatc tgacgatttc ccccgcactc 780
ctggacaaac tgatgaattc aaccgaaccc tttccccgtg tcttggatcc tgtctcggcg 840
aaaaaggaag ccggcgataa aatttcatac atctctgacg agtctaagtt ccgttttgat 900
ctgaatgaag acgctatggc aaccgaaaaa ctgtcagaag gcatccgtaa gttctccgcc 960
gacatcgtta ccctttttga cctcattgaa aagaaagtga cagcc 1005
<210> 134
<211> 1011
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 134
atgtctgaaa atgcagcaaa caacatcatg gaaactaaga tttgcaccga tgccatcgtg 60
tccgagcttc aaaagaaaaa ggtacatttg ttctactgcc tggaatgcga agaactcgct 120
cgaaatatcg cagccgagtc cgatcatatc acccttcaaa gcattaactg gcgctccttc 180
gcagacggtt tccctaacct cttcatcaat aacgcacacg atatccgcgg ccagcacgtc 240
gcgttccttg caagcttcag ctccccagcc gttatctttg aacaaattag cgtgatttat 300
ttgcttcccc gcctcttcgt tgcctcattt actcttgtac ttccgttttt cccaacaggc 360
tccttcgaac gcatggaaga agagggagat gtcgcgaccg ctttcaccat ggctcgaatc 420
gtgagcaaca tcccgatctc ccgcggcgga cctactagcg ttgtgattta tgatatccac 480
gctctccaag aacgctttta ttttgccgac caggtcctgc ctttgttcga gactggcatc 540
cctcttttga cgaaacgtct gcaacaattg cccgaaaccg aaaaggtcat cgtggcattc 600
ccggatgatg gtgcctggaa gcgctttcac aaactcctgg atcattaccc caccgttgtc 660
tgtaccaaag tccgcgaggg tgacaagcgt attgtacgac ttaaagaggg caacccagct 720
ggctgccacg ttgtgatcgt ggatgacttg gtgcagtccg gtggaaccct tatcgaatgt 780
cagaaagtcc tcgcggcgca cggcgctgtt aaggtttccg catacgttac tcacggagtt 840
ttcccaaagt cttcctggga acgcttcact cataagaaaa atggcctgga agaggcattc 900
gcttattttt ggattacaga ctcgtgcccg cagactgtta aggctattgg taataaggct 960
ccattcgagg tcctcagcct ggctggctca attgctgatg ctctccaaat c 1011
<210> 135
<211> 1251
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 135
atggcccttc ttcccttctt cgacctcacc aacttcgagt ccgatgcttc cgaggaactg 60
ggttggctta aatacgtcgg ccgagtacag acacgtgtgt ttccacagca ttttaaggac 120
aatttggaaa aagtgcgcaa aatctccgag actattgatg tcatcgtaga taccaccgct 180
gagttgggcc cggaagcatg tgtgaatctg ctcaatgctg gtgcattggc aatcttggtg 240
aacgaggaga tgcttaatga gttggcagac atctcaccta accgccttgt gttgaagacc 300
gataccaccg atatcggaaa aatcgaaaag ctgtcccagg ttgccggttc gatccagtgg 360
attggaagcg cagagaacta cccaccagat tttttcgaac gtgcatcgaa gatcatccat 420
aaggccgtca tgcctgaagg cggcggacgt actctgtacc tggaattccc agagcaacca 480
tcaatggagg tcctgaaaag cttttcggtg cacagcgttg ttcctgtgct ttcgtcctct 540
ttcctcaccg taaagcccgc agaagaacct aagaaactct ccctggcaga tctcatcctc 600
atctccgcga ataccgaccg cgaggatggc cttttctcaa cactggtggt caacgagctg 660
ggaatcgcac tcggcctcgt gtattcctcc aaagagtcgg tcgcagaatc cttgaagacg 720
ggtactggag tgtaccaatc ccgtaagcga ggactctggt acaagggcgc cagctccggc 780
gcagtccaac acctcattca cattgacgtt gattgcgacg aggattgcct gcgcttcgtg 840
gtttatcaaa ccggaaaagg cttctgccac ctggacactc tgcattgctt tggtcaggcc 900
agcggcttgt gtcagctcga aaaaacgctc atcgatcgta aaaacaatgc gcctgagggc 960
agctacacgg cccgcctgtt ctccgatccc aagcttcttc gagcgaagat tatggaggaa 1020
gccgaggagc tgtgtgatgc caccacgaag gaaaacgtga tctgggaaat ggcggatttg 1080
atgtattttg ccattactcg ttgtgtcggc tccggtgtct cactcaacga cattagccgt 1140
catctggacc tgaagcaccg caaagtgaca cgccgtaaag gtgatgcgaa agtagcatgg 1200
caagagaaat tgaaggataa gggtggcgtg gcaaatacat cctatactgc g 1251
<210> 136
<211> 1005
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 136
atgcactccc accactcgca ctcgggagat tactccgctc atggcaccga tcccctcgat 60
tccgttgtcg atcaagtcgt gaacttgaat ttccacactt actgcttgac agagcatatc 120
ccccgtattg aagccaaatt catttacccg gaggaacagt cgttgggtaa gaacccagag 180
gaagttatta ctaagctcga gacgtccttc aagaatttca tgtcccacgc tcaggagatc 240
aagacccgtt atgccgaccg cccagacgtg cgcaccaaat tcattatcgg tatggagatc 300
gaaagctgtg acatggctca catcgagtat gccaaacgct tgatgaagga aaacaatgac 360
atcctcaagt tctgcgttgg atccgtacac cacgtgaatg gcatccccat tgattttgat 420
cagcagcagt ggtacaactc cctccattcc tttaacgata acctgaagca cttcttgttg 480
tcttactttc aatctcagta tgagatgctc attaacatta agcctctggt ggtcggtcat 540
ttcgacctgt acaaactctt cctgccgaat gacatgttgg ttaatcaaaa atctggtaac 600
tgtaacgaag aaaccggtgt acccgtcgca tcccttgatg ttatttcaga atggcctgag 660
atctacgatg cagtggttcg taaccttcag tttatcgatt catacggcgg cgctatcgaa 720
attaacacct ccgcgttgcg caagcgtctg gaggaaccat acccctctaa aaccctgtgc 780
aacttggtca aaaagcattg cggttcccgc tttgtgctga gcgacgatgc acacggagtg 840
gcacaggtcg gtgtgtgcta tgacaaagtt aagaagtaca ttgtggacgt gctccaactc 900
gaatacatct gttatctgga agaatcacaa tctccagaaa atctgctcac tgtcaagcga 960
ttgccaatct cccaatttgt taatgaccca ttttgggcga acatc 1005
<210> 137
<211> 1542
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 137
atgtctacca acaccactcc ttcctcctgg accaatcctt tgcgtgatcc acaggataag 60
cgtctcccgc gcattgctgg tccttccggt atggttatct tcggtgtaac tggtgatctg 120
gctcgtaaga agctgttgcc agcgatctat gacctggcca atcgcggcct gctcccccca 180
ggcttcagcc tggtcggtta cggccgccgc gagtggagca aggaggactt cgagaaatat 240
gtgcgcgatg cagcgtcagc cggtgcacgt accgaatttc gtgagaacgt ttgggaacgc 300
ctcgcggaag gtatggaatt cgttcgtggt aatttcgacg atgacgccgc ttttgataat 360
ctcgccgcga ccctcaaacg cattgacaaa acgcgcggta ctgcaggtaa ttgggcatac 420
tatctcagca ttccccccga tagctttgcg gcagtatgcc accagctgga acgctccggt 480
atggccgaat ccaccgagga ggcttggcgt cgcgtcatca tcgagaagcc attcggtcac 540
aacctggaat ccgcgcacga acttaaccag ttggtgaatg ccgtgtttcc agaatctagc 600
gtgttccgca tcgatcacta ccttggaaag gaaaccgtgc aaaacatcct tgctctgcgc 660
ttcgctaacc agttgtttga gccgctgtgg aactcaaatt acgttgacca cgtgcagatc 720
actatgacgg aggatatcgg cttgggcggt cgcgccggct actacgacgg tattggagca 780
gcgcgtgacg tgattcaaaa ccatctcatc cagctgctcg cactggttgc aatggaagaa 840
ccgatttcat tcgtcccagc tcaactgcag gctgaaaaga tcaaagtcct gtctgcgaca 900
aagccttgct accctctcga caagacctcg gcacgcggac aatatgcagc cggctggcag 960
ggctccgaac tggttaaggg actccgcgaa gaggatggct ttaacccaga gtccacaacc 1020
gagactttcg ctgcatgcac acttgagatc acatcacgcc gttgggctgg cgtaccgttc 1080
tacctgcgta cgggcaagcg cctgggccgc cgcgtaaccg agatcgcagt tgtgttcaag 1140
gatgctcccc accagccatt cgatggtgat atgaccgttt ctctgggtca aaacgctatt 1200
gtgattcgcg tacagcctga tgaaggcgtt ttgatccgtt tcggttcaaa agtgcctggc 1260
tccgcaatgg aggtacgcga tgtcaatatg gattttagct actctgagtc atttaccgag 1320
gaatcgccag aagcatacga acgcctgatc ctggatgcac tccttgatga gtcttcgctc 1380
ttccctacca acgaagaggt agaactttcc tggaaaattc tggacccaat tctggaagca 1440
tgggatgcag atggcgaacc tgaggactat ccggcaggca cgtggggccc taagtccgcg 1500
gacgaaatgt tgagccgcaa cggccacaca tggcgtcgtc ca 1542
<210> 138
<211> 933
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 138
atgtctgagc tggatgctaa gctgaacaag ctcggagtgg accgaattgc tatctcgccc 60
tacaagcagt ggactcgagg atacatggag cccggcaaca tcggaaatgg atacgtcacg 120
ggacttaagg tcgatgcagg tgtgcgagac aagtccgacg acgacgtcct ggacggaatt 180
gtttcctacg atagagcaga gactaagaac gcttacattg gacaaatcaa catgaccaca 240
gctagctctt tcactggcgt tcaaggccga gtgatcggat acgacattct tcgatctccc 300
gaggtcgata aggccaagcc tctgtttacc gagacccagt gggatggttc cgagcttcct 360
atctacgacg ctaagcctct tcaggatgct ctggttgagt actttggtac tgagcaggat 420
cgtcgtcact acccagctcc cggttccttc atcgtttgtg ctaacaaagg cgtcaccgcc 480
gaacgaccaa agaacgacgc cgatatgaag cccggtcaag gatacggcgt ttggtccgcc 540
atcgctatta gcttcgctaa ggaccctacc aaggactcct ctatgttcgt tgaggacgct 600
ggtgtgtggg agacccctaa cgaggacgag ctgcttgagt acctcgaggg tagacgaaag 660
gccatggcta agtccattgc agaatgtggt caggatgccc acgcctcgtt cgaatcctcc 720
tggattggtt tcgcatacac aatgatggag ccaggccaga ttggtaacgc tattaccgtt 780
gccccctacg tgtctctgcc cattgattcc atccctggtg gctctatcct tactcctgac 840
aaggatatgg agatcatgga gaacctcacc atgccagagt ggctcgaaaa aatgggctac 900
aagtctctct ctgctaacaa cgcccttaag tac 933
<210> 139
<211> 1161
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 139
atgtcgctct ccccgctcga ccaaaaccgc atcgaaagct tctggcagta ttgtctccaa 60
caccagtatt ttaatttggc atatccggaa agcgctgatt ttgattacgc acccttgcat 120
cgctttctgc gcttttccat caacaattgc ggcgattgga acgaaagctc gaactacctg 180
ttgaactctt tcgactttga gcgcgaagtg atgcacttct tcgcagaact gttccacatt 240
ccgttcgacg agtcgtgggg ttacgtgacg aacggcggta cagaaggaaa catgttcggt 300
tgctatctgg ctcgcgaact cttccctgat gcgaccctct actactccaa agattcccac 360
tattccgttg ccaagatcat caaactcttg cgcattaagt cccgcgcagt ggattccctc 420
ccttccggag aaatcgacta cgacgatctc gtggcgaaga tccaacagga ccaggaacga 480
caccctatcg tctttgtcaa tgtaggcact acgatgaaag gtgcggtgga tgacatcggt 540
gtgatccagc ataagcttgc agaagcaggt atcccacgtc aagactacta cctgcacgca 600
gacgcagccc tctcaggtat gatcctccct tttgtggacg caccgcagcc gtattcattt 660
gctgatggca ttgatagcat ctccgtctca ggacataaga tgattggctc tccgatgcct 720
tgcggcattg tactggcgaa gcgctccaac gtgtctcgaa tctccgttga gatcgactac 780
atttccgcaa aggatcagac tatttctggc tctcgaaatg gtcacacgcc catgatgctc 840
tgggcagcta tcaaatcccg tcccctggct gaatggcgcc gtaaagtccg ccattgcctg 900
gatatggccc aatatgccat cgaccgcttg caagctgcag gcatccaagc atggcgctgt 960
aagaacagca ttacggtagt cttcccttcc ccctccgaac cagtgtgcga taaacacggt 1020
ctcgctcgct ccggaggcac cgctcacctc atcaccactc cacaccacca cgattcccag 1080
cgcctcgatc gcctgctgga cgacatcgta caggaccttg gagcgatgac ggcaccagcg 1140
ggtgctacta tgtctgccgc g 1161
<210> 140
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 140
atgctgaaga ttgccgtgcc gaataagggc tctcttagcg aacgtgcgat ggagattctc 60
gccgaagctg gctatgccgg ccgcggcgac tcgaaatcgc tgaacgtgtt tgacgaagca 120
aataacgttg aatttttctt ccttcgaccg aaagacattg caatctatgt ggctggcggc 180
cagcttgatc tgggaatcac gggacgcgac ttggcacgcg actcacaggc agatgttcac 240
gaagtcttga gcttgggttt cggctcgtcg actttccgct acgcggcacc agctgatgag 300
gaatggtcca ttgaaaaact cgatggtaaa cgcattgcta ccagctaccc taatctcgtt 360
cgcgatgatc tcgcagcacg tggactcagc gccgaggtgt tgcgcttgga cggagccgtg 420
gaagtctcca tcaagctcgg cgtggctgac gcgatcgctg atgtggtgtc aaccggccgc 480
acactccgtc aacagggcct ggctcctttc ggtgaggtgc tttgtacctc cgaagcggtg 540
attgtcggtc gtaaggatga gaaggtgaca cccgagcagc agatcttgtt gcgacgaatt 600
cagggtatct tgcatgcgca gaacttcctt atgctggatt acaaggttga tcgagacaat 660
ctcgatgcag ctactgcagt tacccctggc ttctccggcc ctgccgtatc accccttgcg 720
cgtgataatt gggtcgcagt gcgcgcaatg gttcctcgtc gctccgcgaa cgctatcatg 780
gataaactgg ctggactggg tgccgaagct atccttgcta gcgaaatccg catcgcgcga 840
atc 843
<210> 141
<211> 843
<212> DNA
<213> 人工
<220>
<223> 密码子优化和/或突变序列
<400> 141
atgctaaaga tagcagtacc caacaaaggt agcctaagcg agagagctat ggaaatcttg 60
gctgaggctg gctatgctgg gagaggagac tccaaatctc taaatgtgtt tgacgaggct 120
aataatgttg agtttttttt tcttagacca aaggatatcg ccatatatgt cgcgggagga 180
cagctagacc taggcatcac cggcagggat ctagccaggg acagccaagc agatgtccac 240
gaggtcctgt ctttaggatt tggatcttct accttcaggt atgcagcccc tgccgatgaa 300
gagtggtcta tcgagaagct agatggcaaa cgtatcgcca caagttatcc caatttggtc 360
agggatgatc tagccgcgag aggtttatca gccgaggtcc tgagacttga tggggccgtc 420
gaggttagta tcaaactagg tgttgcggat gcgattgcag acgtggtatc caccggaagg 480
acgctacgtc aacaagggct tgccccattc ggtgaggtgc tgtgcacatc cgaggctgtt 540
atcgtgggaa gaaaagacga aaaggttaca cctgagcagc agatattact aagaaggata 600
cagggaattc tgcacgctca aaactttcta atgctggatt ataaggttga tagggataac 660
ctagacgccg ccacagccgt tactcctggt ttctccggtc ccgctgtcag tcctctagct 720
agggacaact gggtcgccgt cagggctatg gttccaagac gtagcgctaa cgcaataatg 780
gacaaattgg cgggccttgg ggccgaagca atactggcga gcgaaatcag gattgcacgt 840
att 843
<210> 142
<211> 55
<212> PRT
<213> 人工
<220>
<223> 溶解性标签
<400> 142
Met Gln Tyr Lys Leu Ala Leu Asn Gly Lys Thr Leu Lys Gly Glu Thr
1 5 10 15
Thr Thr Glu Ala Val Asp Ala Ala Thr Ala Glu Lys Val Phe Lys Gln
20 25 30
Tyr Ala Asn Asp Asn Gly Val Asp Gly Glu Trp Thr Tyr Asp Asp Ala
35 40 45
Thr Lys Thr Phe Thr Val Thr
50 55
<210> 143
<211> 317
<212> PRT
<213> 人工
<220>
<223> 突变的
<400> 143
Met Ser Asn Glu Tyr Gly Asp Lys Asn Leu Lys Ile Phe Ser Leu Asn
1 5 10 15
Ser Asn Pro Glu Leu Ala Lys Glu Ile Ala Asp Asn Val Gly Val Gln
20 25 30
Leu Gly Lys Cys Ser Val Thr Arg Phe Ser Asp Gly Glu Val Gln Ile
35 40 45
Asn Ile Glu Glu Ser Ile Arg Gly Cys Asp Cys Tyr Ile Ile Gln Ser
50 55 60
Thr Ser Ala Pro Val Asn Glu His Ile Met Glu Leu Leu Ile Met Val
65 70 75 80
Asp Ala Leu Lys Arg Ala Ser Ala Lys Thr Ile Asn Ile Val Ile Pro
85 90 95
Tyr Tyr Gly Tyr Ala Arg Gln Asp Arg Lys Ala Arg Ser Arg Glu Pro
100 105 110
Ile Thr Ala Lys Leu Phe Ala Asn Leu Leu Glu Thr Ala Gly Ala Thr
115 120 125
Arg Val Ile Ala Leu Asp Ile His Ala Pro Gln Ile Gln Gly Phe Phe
130 135 140
Asp Ile Pro Ile Asp His Leu Met Gly Val Pro Ile Leu Gly His Tyr
145 150 155 160
Phe Glu Gly Lys Asp Leu Lys Asp Ile Val Ile Val Ser Pro Asp His
165 170 175
Gly Gly Val Thr Arg Ala Arg Lys Leu Ala Asp Arg Leu Lys Ala Pro
180 185 190
Ile Ala Ile Ile Asp Lys Arg Arg Pro Arg Pro Asn Glu Val Glu Val
195 200 205
Met Asn Ile Val Gly Asn Val Glu Gly Lys Thr Ala Ile Leu Ile Asp
210 215 220
Asp Ile Ile Asp Thr Ala Gly Thr Ile Thr Leu Ala Ala Asn Ala Leu
225 230 235 240
Val Glu Asn Gly Ala Ala Glu Val Tyr Ala Cys Cys Thr His Pro Val
245 250 255
Leu Ser Gly Pro Ala Val Glu Arg Ile Asn Asn Ser Lys Ile Lys Glu
260 265 270
Leu Val Val Thr Asn Ser Ile Lys Leu Pro Glu Glu Lys Lys Ile Glu
275 280 285
Arg Phe Lys Gln Leu Ser Val Gly Pro Leu Leu Ala Glu Ala Ile Ile
290 295 300
Arg Val His Glu Lys Gln Ser Val Ser Tyr Leu Phe Ser
305 310 315

Claims (15)

1.一种表达非天然的组氨酸脱羧酶的经改造的微生物细胞，其中所述经改造的微生物细胞产生组胺。
2.权利要求1的经改造的微生物细胞，其中所述经改造的微生物细胞包括一种或多种上游组胺途径酶的提高的活性，所述提高的活性为相对于对照细胞的提高。
3.权利要求2的经改造的微生物细胞，其中所述一种或多种上游组胺途径酶选自ATP磷酸核糖基转移酶、磷酸核糖基-ATP焦磷酸酶、磷酸核糖基-AMP环化水解酶、5’ProFAR异构酶、咪唑-甘油磷酸合酶、咪唑-甘油磷酸脱水酶、组氨醇-磷酸氨基转移酶、组氨醇-磷酸磷酸酶、组氨醇脱氢酶和磷酸核糖焦磷酸激酶。
4.权利要求1的经改造的微生物细胞，其中所述经改造的微生物细胞包括一种或多种消耗一种或多种组胺途径前体的酶的降低的活性，所述降低的活性为相对于对照细胞的降低。
5.权利要求1的经改造的微生物细胞，其中所述经改造的微生物细胞额外地表达反馈失调的葡萄糖-6-磷酸脱氢酶或反馈失调的ATP磷酸核糖基转移酶。
6.一种经改造的微生物细胞，其中所述经改造的微生物细胞包括用于表达非天然的组氨酸脱羧酶的方式，其中所述经改造的微生物细胞产生组胺。
7.权利要求6的经改造的微生物细胞，其中所述经改造的微生物细胞还包括用于表达葡萄糖-6-磷酸脱氢酶或反馈失调的ATP磷酸核糖基转移酶的方式。
8.权利要求1的经改造的微生物细胞，其中所述经改造的微生物细胞包括真菌细胞。
9.权利要求1的经改造的微生物细胞，其中所述非天然的组氨酸脱羧酶包括与来自色杆菌属种LK1或来自鲍氏不动杆菌菌株AB0057的组氨酸脱羧酶具有至少70％氨基酸序列同一性的组氨酸脱羧酶。
10.权利要求1的经改造的微生物细胞，其中所述经改造的微生物细胞为细菌细胞。
11.权利要求1的经改造的微生物细胞，其中所述经改造的微生物细胞包括一种或多种上游组胺途径酶的提高的活性，所述提高的活性为相对于对照细胞的提高，其中所述一种或多种上游组胺途径酶包括ATP磷酸核糖基转移酶和咪唑-甘油磷酸脱水酶。
12.根据权利要求1中任一项的经改造的微生物细胞的培养物。
13.权利要求12的培养物，其中所述培养物包含水平为至少20mg/L培养基的组胺。
14.培养根据权利要求1的经改造的微生物细胞的方法，所述方法包括在适合产生组胺的条件下培养所述细胞。
15.使用经改造以产生组胺的微生物细胞来制备组胺的方法，所述方法包括：
(a)在微生物细胞中表达非天然的组氨酸脱羧酶；
(b)在适合的培养基中，在允许微生物细胞产生组胺的条件下培养微生物细胞，其中所述组胺被释放到培养基中；和
(c)从培养基中分离组胺。
CN201980036608.0A 2018-04-20 2019-04-19 通过发酵产生组胺的经改造的生物合成途径 Pending CN112368387A (zh)

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