CN113122519B - 一种耐热6-磷酸氨基葡萄糖磷酸酶突变体及其应用 - Google Patents

一种耐热6-磷酸氨基葡萄糖磷酸酶突变体及其应用 Download PDF

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CN113122519B
CN113122519B CN201911408177.4A CN201911408177A CN113122519B CN 113122519 B CN113122519 B CN 113122519B CN 201911408177 A CN201911408177 A CN 201911408177A CN 113122519 B CN113122519 B CN 113122519B
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游淳
孟冬冬
周伟
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Tianjin Institute of Industrial Biotechnology of CAS
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Abstract

本发明公开了一种6‑磷酸氨基葡萄糖磷酸酶突变体，属于基因工程和酶工程领域。本发明采用理性设计的方法，经过多轮定点突变，提供了一系列来源于多形拟杆菌的6‑磷酸氨基葡萄糖磷酸酶突变体。与野生型6‑磷酸氨基葡萄糖磷酸酶相比，突变体具有明显提高的热稳定性和催化活力。本发明所示6‑磷酸氨基葡萄糖磷酸酶将在氨基葡萄糖的工业生产中显示出巨大的潜在价值。

Description

一种耐热6-磷酸氨基葡萄糖磷酸酶突变体及其应用
技术领域
本发明涉及一种6-磷酸氨基葡萄糖磷酸酶突变体，具体的说是一种耐热6-磷酸氨基葡萄糖磷酸酶突变体及其应用，属于基因工程和酶工程领域。
背景技术
氨基葡萄糖是一种天然存在的功能性氨基单糖，它是葡萄糖分子中2位羟基被氨基取代后的化合物。因为氨基葡萄糖的安全性和功能多样性，氨基葡萄糖及其衍生物在医药、食品、化妆品等领域具有重要用途。
传统氨基葡萄生产方法为甲壳素水解法，但是受到原料供应限制，生产过程需要的高浓度盐酸会对环境造成严重污染，而且虾蟹壳来源的氨基葡萄糖会造成一些人群的过敏反应。通过工程菌发酵葡萄糖生产氨基葡萄糖是一种不受原料来源限制、环境污染低、对人体安全的生产方法，但是存在一些缺点，如发酵产物通常为乙酰氨基葡萄糖、工程菌代谢工程改造困难、产生代谢副产物等。针对上述困境，我们前期开发了一种低成本、低污染的通过体外多酶系统催化将淀粉转化为氨基葡萄糖的新方法(中国专利申请号：201810772487.3)。在酶资源挖掘、反应体系优化基础上，该体外多酶体系在37℃下催化10g/L经异淀粉酶处理的可溶性淀粉生产7.93g/L氨基葡萄糖。在工业生产中，使用热稳定性高的酶在高温条件下进行反应可以降低微生物污染风险、降低酶的制备成本、降低反应溶液粘度从而提高传质效率。然而，酶的热稳定性，尤其是6-磷酸氨基葡萄糖磷酸酶(6-phosphate glucosamine phosphatase，GlmP)，限制了该氨基葡萄糖生产新方法的工业化应用。
因此，亟需对6-磷酸氨基葡萄糖磷酸酶进行热稳定性改造，推动体外多酶体系生产氨基葡萄糖的工业化。
发明内容
本发明的目的在于提供一种耐热6-磷酸氨基葡萄糖磷酸酶突变体及其应用。为实现上述目的，第一方面，本发明提供一种6-磷酸氨基葡萄糖磷酸酶突变体，采取的技术方案为：以氨基酸序列为SEQ ID NO：2所示的野生型GlmP作为出发蛋白，通过理性设计对其进行多轮定点突变，所述突变体与野生型6-磷酸氨基葡萄糖磷酸酶氨基酸序列SEQ ID NO：2具有至少90％同一性，且所述突变体具有催化6-磷酸氨基葡萄糖转化为氨基葡萄糖的活性。
进一步地，所述突变体与SEQ ID NO：2具有至少93％、至少94％、至少96％、至少97％、至少98％或至少99％的序列同一性。
进一步地，所述突变体包括在对应于SEQ ID NO：2上的一个或多个位点上的突变，所述的位点对应于SEQ ID NO：2的氨基酸1至258中的位点9、17、22、25、56、78、81、100、123、126、127、142、148、164、167、190、197、200、246。
进一步地，所述突变体包括在对应于SEQ ID NO：2上的以下任一种位点的突变：
在9、17、22、25、56、78、81、100、123、126、127、142、148、164、167、190、197、200、246位任一位点的单突变；
在17位和126位和127位和142位的组合突变；
在17位和22位和126位和127位和142位的组合突变；
在17位和100位和126位和127位和142位的组合突变；
在17位和126位和127位和142位和164位的组合突变；
在17位和126位和127位和142位和197位的组合突变；
在17位和100位和126位和127位和142位和164位的组合突变；
在17位和126位和127位和142位和164位和197位的组合突变；
在17位和100位和126位和127位和142位和164位和197位的组合突变；
在17位和22位和100位和126位和127位和142位和164位的组合突变；
在9位和17位和22位和100位和126位和127位和142位和164位的组合突变；
在17位和22位和25位和100位和126位和127位和142位和164位的组合突变；
在17位和22位和100位和123位和126位和127位和142位和164位的组合突变；
在17位和22位和100位和126位和127位和142位和164位和190位的组合突变；
在17位和22位和100位和126位和127位和142位和164位和200位的组合突变；
在17位和22位和100位和126位和127位和142位和164位和246位的组合突变；
在17位和22位和和25位100位和126位和127位和142位和164位和246位的组合突变；
在17位和22位和100位和126位和127位和142位和164位和190位和200位的组合突变；
在17位和22位和100位和126位和127位和142位和164位和190位和200位和246位的组合突变；
在17位和22位和25位和100位和126位和127位和142位和164位和190位和200位和246位的组合突变；
在17位和22位和25位和56位和100位和126位和127位和142位和164位和246位的组合突变；
在17位和22位和25位和78位和100位和126位和127位和142位和164位和246位的组合突变；
在17位和22位和25位和81位和100位和126位和127位和142位和164位和246位的组合突变；
在17位和22位和25位和100位和126位和127位和142位和148位和164位和246位的组合突变；
在17位和22位和25位和100位和126位和127位和142位和164位和167位和246位的组合突变；
在17位和22位和25位和56位和81位和100位和126位和127位和142位和164位和246位的组合突变；
优选地，所述突变体对应于SEQ ID NO：2上的位点9的氨基酸由缬氨酸(V)突变为异亮氨酸(I)；位点17的氨基酸由谷氨酸(E)突变为赖氨酸(K)；位点22的氨基酸由丝氨酸(S)突变为脯氨酸(P)；位点25的氨基酸由丝氨酸(S)突变为苏氨酸(T)；位点56的氨基酸由缬氨酸(V)突变为异亮氨酸(I)；位点78位的氨基酸由精氨酸(R)突变为酪氨酸(Y)；位点81位的氨基酸由丙氨酸(A)突变为脯氨酸(P)；位点100位的氨基酸由丙氨酸(A)突变为脯氨酸(P)；位点123位的氨基酸由甘氨酸(G)突变为酪氨酸(Y)；位点126位的氨基酸由谷氨酸(E)突变为天冬酰胺(N)、位点127位的氨基酸由组氨酸(H)突变为精氨酸(R)、位点142位的氨基酸由赖氨酸(K)突变为缬氨酸(V)、位点148位的氨基酸由半胱氨酸(C)突变为缬氨酸(V)、位点164位的氨基酸由丝氨酸(S)突变为脯氨酸(P)；位点167位的氨基酸由丝氨酸(S)突变为异亮氨酸(I)、位点197位的氨基酸由精氨酸(R)突变为甘氨酸(G)、位点190位的氨基酸由丝氨酸(S)突变为苯丙氨酸(F)、位点200位氨基酸由缬氨酸(V)突变为亮氨酸(L)、位点246位氨基酸由丝氨酸(S)突变为天门冬氨酸(D)。
优选地，所述突变体的由包含如SEQ ID NO.4、SEQ ID NO.6、SEQ ID NO.8、SEQ IDNO.10、SEQ ID NO.12、SEQ ID NO.14、SEQ ID NO.16、SEQ ID NO.18、SEQ ID NO.20、SEQ IDNO.22、SEQ ID NO.24、SEQ ID NO.26、SEQ ID NO.28、SEQ ID NO.30、SEQ ID NO.32、SEQ IDNO.34、SEQ ID NO.36、SEQ ID NO.38、SEQ ID NO.40、SEQ ID NO.42、SEQ ID NO.44、SEQ IDNO.46、SEQ ID NO.48、SEQ ID NO.50、SEQ ID NO.52、SEQ ID NO.54、SEQ ID NO.56、SEQ IDNO.58和SEQ ID NO.60所示的序列编码。
优选地，所述6-磷酸氨基葡萄糖磷酸酶来源于多形拟杆菌(Bacteroidesthetaiotaomicron)。
第二方面，本发明提供一种多核苷酸，所述多核苷酸包含编码第一方面任一所述的突变体的核苷酸序列。
第三方面，本发明提供一种宿主细胞，所述宿主细胞包含第二方面所述的多核苷酸。
第四方面，本发明提供一种第一方面所述突变体，第二方面所述多核苷酸，第三方面所述宿主细胞在制备氨基葡萄糖中的应用。
本发明的有益效果：
1.通过构建一系列热稳定性提高的GlmP突变体，提高GlmP热稳定性性能，实现氨基葡萄糖在高温环境下的合成。
2.氨基葡萄糖的高温合成有利于降低微生物污染风险、降低酶的制备成本、降低反应溶液粘度从而提高传质效率，通过提供一系列热稳定性提高的GlmP突变体，为氨基葡萄糖的工业化生产提供保障。
附图说明
图1 50℃下，野生型6-磷酸氨基葡萄糖磷酸酶及其突变体的热稳定性。
图2 50℃下体外多酶催化IA处理过的可溶性淀粉合成氨基葡萄糖的反应进程曲线。
具体实施方式
下面结合具体实施例来进一步描述本发明，本发明的优点和特点将会随着描述而更为清楚。但是应理解所述实施例仅是范例性的，不对本发明的范围构成任何限制。本领域技术人员应该理解的是，在不偏离本发明的精神和范围下可以对本发明技术方案的细节和形式进行修改或替换，但这些修改或替换均落入本发明的保护范围。
本发明实施例中用到如下材料
麦芽糊精，sigma公司产品，产品编号：419672；
pET20b载体，Novagen，Madison，WI；
大肠杆菌表达菌BL21(DE3)，Invitrogen，Carlsbad，CA；
GlmP突变体的载体构建方法：利用Phusion定点突变方法，设计常规引物，以含有野生型GlmP的载体pET20b-glmp或含有每轮最优突变体的载体为模板，进行PCR扩增。PCR反应体系均为：2﹡PrimerStar mixture 25μL，正向引物(10μM)1μL，反向引物(10μM)1μL，模板DNA 1μL，加入双蒸水22μL。PCR反应扩增条件均为：98℃预变性45s；随后98℃15s，55℃15s，72℃45s进行30个循环，最后72℃保温10min。将PCR产物经过Dpn I消化30min后，利用PCR产物纯化试剂盒进行纯化。纯化后的PCR产物3μL，与T4 PNK 1μL、10﹡T4PNK buffer 1μL、100mM ATP 1μL、双蒸水4μL混匀，37℃磷酸化反应2h。取磷酸化的PCR产物溶液10μL与快速连接酶1μL、快速连接酶缓冲液10μL混匀，25℃温浴30min，进行平末端连接。连接产物分别转入大肠杆菌E.coli TOP10的感受态细胞中，涂布到含有抗生素的LB固体培养基平板上，培养过夜。挑取单菌落于LB液体培养基中培养过夜后提取质粒并进行测序验证。
GlmP突变体的表达：挑取含有测序正确突变体基因载体的BL21(DE3)菌株接种LB液体培养基，在37℃，200rpm下培养到OD600nm为0.6-0.8，加入终浓度0.1mM IPTG诱导，25℃继续培养12h。收集菌体，经超声破碎获得突变体蛋白粗酶液，经镍柱纯化、超滤浓缩和置换缓冲液，获得电泳纯突变体蛋白。
GlmP突变体的热稳定性测定方法：将突变体蛋白稀释到终浓度1g/L(100mMHEPES，pH 7.5)，置于50℃水浴中，不同时间取样，测定残余酶活。以温浴0h蛋白的比酶活作为100％。6-磷酸氨基葡萄糖磷酸酶比酶活测定方法：在含有10mM氯化镁的100mM HEPES缓冲液(pH 7.5)中，添加终浓度10mM的6-磷酸氨基葡萄糖(GlcN6P)，37℃反应10min，测定产生的无机磷的量。
6-磷酸氨基葡萄糖磷酸酶是以6-磷酸氨基葡萄糖作为底物进行催化脱磷酸反应，从而生成6-磷酸氨基葡萄糖和无机磷的酶。
实施例1第一轮定点突变及组合突变
以野生型GlmP的晶体结构(PDB ID：3R4C)为基础，经B-因子分析，确定10个最不稳定的氨基酸位点，然后使用Rosetta程序，对上述10个位点做虚拟突变，设计突变体。以含有如SEQ ID NO：1所示的经过密码子优化后的野生型GlmP编码基因的载体pET20b-glmP为模板，进行PCR扩增，构建GlmP突变体的表达载体。经蛋白表达，获得突变体蛋白，测定突变体的热稳定性。如SEQ ID NO：2所示的野生型GlmP在50℃下的半衰期小于1min。本轮突变获得4个点突变后热稳定性提高的突变体：GlmP-E17K、GlmP-E126N、GlmP-H127R和GlmP-K142V，在50℃下半衰期分别为3.2min、5.8min、6.2min和8.2min。对上述位点进行组合突变，获得组合突变体GlmP-E17K/E126N/H127R/K142V，在50℃下半衰期提高到16min。突变体GlmP-E17K的编码核酸序列为SEQ ID NO.3所示，氨基酸序列为SEQ ID NO.4所示；突变体GlmP-E126N的编码核酸序列为SEQ ID NO.5所示，氨基酸序列为SEQ ID NO.6所示；突变体GlmP-H127R的编码核酸序列为SEQ ID NO.7所示，氨基酸序列为SEQ ID NO.8所示；突变体GlmP-K142V的编码核酸序列为SEQ ID NO.9所示，氨基酸序列为SEQ ID NO.10所示；本轮获得的最优突变体GlmP-E17K/E126N/H127R/K142V的编码核酸序列为SEQ ID NO.11所示，氨基酸序列为SEQ ID NO.12所示，命名为R1。
实施例2第二轮定点突变及组合突变
以R1为出发蛋白，经B-因子分析，确定12个最不稳定的氨基酸位点，然后使用Rosetta程序，对上述位点做虚拟突变，设计突变体。经蛋白表达，获得突变体蛋白，测定突变体的热稳定性。本轮突变获得4个点突变后热稳定性提高的突变体：R1-S22P、R1-A100P、R1-S164P、R1-R197G，在50℃下半衰期分别为35min、55min、68min和55min。对上述位点进行组合突变，分别获得组合突变体R1-A100P/S164P、R1-S164P/R197G、R1-A100P/S164P/R197G和R1-S22P/A100P/S164P，在50℃下半衰期分别为1.88h、55min、1.69h和2.09h。突变体R1-S22P的编码核酸序列为SEQ ID NO.13所示，氨基酸序列为SEQ ID NO.14所示；突变体R1-A100P的编码核酸序列为SEQ ID NO.15所示，氨基酸序列为SEQ ID NO.16所示；突变体R1-S164P的编码核酸序列为SEQ ID NO.17所示，氨基酸序列为SEQ ID NO.18所示；突变体R1-R197G的编码核酸序列为SEQ ID NO.19所示，氨基酸序列为SEQ ID NO.20所示；突变体R1-A100P/S164P的编码核酸序列为SEQ ID NO.21所示，氨基酸序列为SEQ ID NO.22所示；突变体R1-S164P/R197G的编码核酸序列为SEQ ID NO.23所示，氨基酸序列为SEQ ID NO.24所示；突变体R1-A100P/S164P/R197G的编码核酸序列为SEQ ID NO.25所示，氨基酸序列为SEQID NO.26所示；本轮获得的最优突变体R1-S22P/A100P/S164P的编码核酸序列为SEQ IDNO.27所示，氨基酸序列为SEQ ID NO.28所示，命名为R2。
实施例3第三轮定点突变及组合突变
以R2为出发蛋白，经FoldX和Rosetta软件分析，确定8个待突变位点。经蛋白表达，获得突变体蛋白，测定突变体的热稳定性。本轮突变获得6个点突变后热稳定性提高的突变体：R2-V9I、R2-S25T、R2-G123Y、R2-S190F、R2-V200L和R2-S246D，在50℃下半衰期分别为3.85h、5.45h、4.07h、4.28h、3.44h和7.16h。对上述位点进行组合突变，分别获得组合突变体R2-S25T/S246D、R2-S190F/V200L、R2-S190F/V200L/S246D和R2-S25T/S190F/V200L/S246D，在50℃下半衰期分别为7.7h、3.03h、6.16h和4.8h。突变体R2-V9I的编码核酸序列为SEQ ID NO.29所示，氨基酸序列为SEQ ID NO.30所示；突变体R2-S25T的编码核酸序列为SEQ ID NO.31所示，氨基酸序列为SEQ ID NO.32所示；突变体R2-G123Y的编码核酸序列为SEQ ID NO.33所示，氨基酸序列为SEQ ID NO.34所示；突变体R2-S190F的编码核酸序列为SEQ ID NO.35所示，氨基酸序列为SEQ ID NO.36所示；突变体R2-V200L的编码核酸序列为SEQ ID NO.37所示，氨基酸序列为SEQ ID NO.38所示；突变体R2-S246D的编码核酸序列为SEQ ID NO.39所示，氨基酸序列为SEQ ID NO.40所示；突变体R2-S25T/S246D的编码核酸序列为SEQ ID NO.41所示，氨基酸序列为SEQ ID NO.42所示；突变体R2-S190F/V200L的编码核酸序列为SEQ ID NO.43所示，氨基酸序列为SEQ ID NO.44所示；突变体R2-S190F/V200L/S246D的编码核酸序列为SEQ ID NO.45所示，氨基酸序列为SEQ ID NO.46所示；突变体R2-S25T/S190F/V200L/S246D的编码核酸序列为SEQ ID NO.47所示，氨基酸序列为SEQ IDNO.48所示。本轮获得的最优突变体R2-S25T/S246D命名为R3。
实施例4第四轮定点突变及组合突变
以R3为出发蛋白，经FoldX和Rosetta软件分析，确定6个待突变位点。经蛋白表达纯化，获得突变体蛋白，测定突变体的热稳定性。本轮突变获得5个点突变后热稳定性提高的突变体：R3-V56I、R3-R78Y、R3-A81P、R3-C148V和R3-S167I，在50℃下半衰期分别为15.8h、10.4h、15.5h、11.3h和8.3h。对上述位点进行组合突变，获得组合突变体R3-V56I/A81P，在50℃下半衰期为30.3h。突变体R3-V56I的编码核酸序列为SEQ ID NO.49所示，氨基酸序列为SEQ ID NO.50所示；突变体R3-R78Y的编码核酸序列为SEQ ID NO.51所示，氨基酸序列为SEQ ID NO.52所示；突变体R3-A81P的编码核酸序列为SEQ ID NO.53所示，氨基酸序列为SEQ ID NO.54所示；突变体R3-C148V的编码核酸序列为SEQ ID NO.55所示，氨基酸序列为SEQ ID NO.56所示；突变体R3-S167I的编码核酸序列为SEQ ID NO.57所示，氨基酸序列为SEQ ID NO.58所示；本轮获得的最优突变体R3-V56I/A81P的编码核酸序列为SEQ IDNO.59所示，氨基酸序列为SEQ ID NO.60所示，命名为R4。
经过4轮定点突变和组合突变，最优突变体R4在50℃半衰期30.3h，55℃半衰期10h，60℃下半衰期1.9h。突变体R4在37℃下对GlcN6P的比酶活5.4U/mg，较野生型GlmP比酶活3U/mg，提高1.8倍。各轮突变后最优突变体的热稳定性如图1所示。
实施例5高温条件下体外多酶催化可溶性淀粉生产氨基葡萄糖
根据专利(中国专利申请号：201810772487.3)中所述对可溶性淀粉进行异淀粉酶IA处理，提高多酶体系中底物转利率。将含有10g/L经异淀粉酶处理的可溶性淀粉、10mM氯化镁、20mM磷酸二氢钾、200mM氯化铵、100mM HEPES缓冲液(pH 7.5)、2U/mLαGP、2U/mL PGM、3U/mL PGI、2U/mL GlmD、2U/mL GlmP野生型蛋白或2U/mL R4最优突变体的0.5mL反应混合物在50℃下孵育7h。不同时间取样，通过添加高氯酸终止反应，氢氧化钾中和pH至7.0，12000转离心10min，取上清通过高效液相色谱法测定氨基葡萄糖的浓度。所用色谱柱为Agilent ZORBAX SB-C18(300mm)，衍生试剂为邻苯二甲醛(OPA)，流动相为40mM磷酸氢二钠(A相，pH 7.8)和乙腈：甲醇：水＝45:45:10(B相)，流速1mL/min，柱温40℃，所用检测器为紫外检测器，检测波长338nm。含有突变体R4的多酶体系反应进行4h时，氨基葡萄糖的浓度为3.4g/L，转化率为34％(图2)。含有野生型蛋白GlmP的多酶体系没有检测到氨糖产生。
以上，对本发明的实施方式进行了说明。但是，本发明不限定于上述实施方式。凡在本发明的精神和原则之内，所做的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
序列表
<110> 中国科学院天津工业生物技术研究所
<120> 一种耐热6-磷酸氨基葡萄糖磷酸酶突变体及其应用
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gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtggaaca tccggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtaaggagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctga gcggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
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atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
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attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
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cgtaaggagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctga gcggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
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Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
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atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttga aacccataaa 60
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attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
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cgtaaggagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
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gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
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<210> 7
<211> 777
<212> DNA
<213> 人工序列()
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atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttga aacccataaa 60
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attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtggaaag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtaaggagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctga gcggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 8
<211> 258
<212> PRT
<213> 人工序列()
<400> 8
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
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Glu Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
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Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
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Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Lys Glu Cys
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Pro Leu Leu Ser Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
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Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
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Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
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Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
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Val Ile
<210> 9
<211> 777
<212> DNA
<213> 人工序列()
<400> 9
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttga aacccataaa 60
gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtggaaca tccggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctga gcggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 10
<211> 258
<212> PRT
<213> 人工序列()
<400> 10
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Glu Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Ala Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Glu His Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Ser Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 11
<211> 777
<212> DNA
<213> 人工序列()
<400> 11
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctga gcggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 12
<211> 258
<212> PRT
<213> 人工序列()
<400> 12
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Ala Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Ser Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 13
<211> 777
<212> DNA
<213> 人工序列()
<400> 13
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctga gcggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 14
<211> 258
<212> PRT
<213> 人工序列()
<400> 14
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Ala Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Ser Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 15
<211> 777
<212> DNA
<213> 人工序列()
<400> 15
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctga gcggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 16
<211> 258
<212> PRT
<213> 人工序列()
<400> 16
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Ser Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 17
<211> 777
<212> DNA
<213> 人工序列()
<400> 17
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 18
<211> 258
<212> PRT
<213> 人工序列()
<400> 18
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Ala Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 19
<211> 777
<212> DNA
<213> 人工序列()
<400> 19
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctga gcggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatgg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 20
<211> 258
<212> PRT
<213> 人工序列()
<400> 20
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Ala Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Ser Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Gly Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 21
<211> 777
<212> DNA
<213> 人工序列()
<400> 21
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 22
<211> 258
<212> PRT
<213> 人工序列()
<400> 22
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 23
<211> 777
<212> DNA
<213> 人工序列()
<400> 23
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttgca 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatgg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 24
<211> 258
<212> PRT
<213> 人工序列()
<400> 24
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Ala Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Gly Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 25
<211> 777
<212> DNA
<213> 人工序列()
<400> 25
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgagccaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatgg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 26
<211> 258
<212> PRT
<213> 人工序列()
<400> 26
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Ser Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Gly Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 27
<211> 777
<212> DNA
<213> 人工序列()
<400> 27
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 28
<211> 258
<212> PRT
<213> 人工序列()
<400> 28
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 29
<211> 777
<212> DNA
<213> 人工序列()
<400> 29
atgatcaagg tgctgctgct ggatattgat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 30
<211> 258
<212> PRT
<213> 人工序列()
<400> 30
Met Ile Lys Val Leu Leu Leu Asp Ile Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 31
<211> 777
<212> DNA
<213> 人工序列()
<400> 31
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 32
<211> 258
<212> PRT
<213> 人工序列()
<400> 32
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 33
<211> 777
<212> DNA
<213> 人工序列()
<400> 33
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcatata ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 34
<211> 258
<212> PRT
<213> 人工序列()
<400> 34
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Tyr Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 35
<211> 777
<212> DNA
<213> 人工序列()
<400> 35
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttattt ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 36
<211> 258
<212> PRT
<213> 人工序列()
<400> 36
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Phe Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 37
<211> 777
<212> DNA
<213> 人工序列()
<400> 37
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaactg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 38
<211> 258
<212> PRT
<213> 人工序列()
<400> 38
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Leu Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 39
<211> 777
<212> DNA
<213> 人工序列()
<400> 39
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 40
<211> 258
<212> PRT
<213> 人工序列()
<400> 40
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 41
<211> 777
<212> DNA
<213> 人工序列()
<400> 41
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 42
<211> 258
<212> PRT
<213> 人工序列()
<400> 42
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 43
<211> 777
<212> DNA
<213> 人工序列()
<400> 43
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttattt ctgtttgcag attattatcg cgtgaaactg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatagcgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 44
<211> 258
<212> PRT
<213> 人工序列()
<400> 44
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Phe Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Leu Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Ser Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 45
<211> 777
<212> DNA
<213> 人工序列()
<400> 45
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcagtattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttattt ctgtttgcag attattatcg cgtgaaactg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 46
<211> 258
<212> PRT
<213> 人工序列()
<400> 46
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Ser Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Phe Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Leu Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 47
<211> 777
<212> DNA
<213> 人工序列()
<400> 47
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttattt ctgtttgcag attattatcg cgtgaaactg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 48
<211> 258
<212> PRT
<213> 人工序列()
<400> 48
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Phe Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Leu Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 49
<211> 777
<212> DNA
<213> 人工序列()
<400> 49
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcaattcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 50
<211> 258
<212> PRT
<213> 人工序列()
<400> 50
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Ile Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 51
<211> 777
<212> DNA
<213> 人工序列()
<400> 51
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat ttataaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 52
<211> 258
<212> PRT
<213> 人工序列()
<400> 52
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Tyr Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 53
<211> 777
<212> DNA
<213> 人工序列()
<400> 53
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
ccgattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 54
<211> 258
<212> PRT
<213> 人工序列()
<400> 54
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Pro Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 55
<211> 777
<212> DNA
<213> 人工序列()
<400> 55
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct ggtgttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 56
<211> 258
<212> PRT
<213> 人工序列()
<400> 56
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Val Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 57
<211> 777
<212> DNA
<213> 人工序列()
<400> 57
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcagttcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
gccattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgat tgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 58
<211> 258
<212> PRT
<213> 人工序列()
<400> 58
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Val Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Ala Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ile Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile
<210> 59
<211> 777
<212> DNA
<213> 人工序列()
<400> 59
atgatcaagg tgctgctgct ggatgtggat ggcaccttac tgagctttaa aacccataaa 60
gtgccgcaga gcaccattga tgcactgaaa aaagtgcacg acagtggtat taagatcgtt 120
attgcaaccg gccgcgccgc atcagatctt cacgagattg atgcaattcc gtatgatggc 180
gttattgccc tgaatggtgc cgaatgcgtg ctgagagatg gctcagtgat tcgtaaagtt 240
ccgattccgg cacaggattt tcgtaaaagc atggaactgg cacgcgaatt tgattttccg 300
gttgcactgg aactgaacga aggtgtgttt gtgaatcgtc tgaccccgac cgttgaacag 360
attgcaggca ttgtgaatag accggttccg cctgtggtgg atattgaaga aatgtttgag 420
cgtgttgagt gctgtcagct gtgtttttat ttcgatgagg aggcagaaca gaaggttatg 480
ccgctgctgc cgggtctgag cgcaacccgt tggcaccctt tatttgcaga tgttaatgtg 540
gcaggtacca gcaaagcaac cggcttaagc ctgtttgcag attattatcg cgtgaaagtg 600
agcgaaatca tggcatgcgg cgatggtggt aatgatattc cgatgctgaa agcagcaggc 660
attggtgtgg ccatgggcaa tgctagtgaa aaagttcaga gtgtggcaga tttcgtgacc 720
gataccgtgg ataatgatgg cctgtataaa gccctgaaac attttggtgt gatctga 777
<210> 60
<211> 258
<212> PRT
<213> 人工序列()
<400> 60
Met Ile Lys Val Leu Leu Leu Asp Val Asp Gly Thr Leu Leu Ser Phe
1 5 10 15
Lys Thr His Lys Val Pro Gln Ser Thr Ile Asp Ala Leu Lys Lys Val
20 25 30
His Asp Ser Gly Ile Lys Ile Val Ile Ala Thr Gly Arg Ala Ala Ser
35 40 45
Asp Leu His Glu Ile Asp Ala Ile Pro Tyr Asp Gly Val Ile Ala Leu
50 55 60
Asn Gly Ala Glu Cys Val Leu Arg Asp Gly Ser Val Ile Arg Lys Val
65 70 75 80
Pro Ile Pro Ala Gln Asp Phe Arg Lys Ser Met Glu Leu Ala Arg Glu
85 90 95
Phe Asp Phe Pro Val Ala Leu Glu Leu Asn Glu Gly Val Phe Val Asn
100 105 110
Arg Leu Thr Pro Thr Val Glu Gln Ile Ala Gly Ile Val Asn Arg Pro
115 120 125
Val Pro Pro Val Val Asp Ile Glu Glu Met Phe Glu Arg Val Glu Cys
130 135 140
Cys Gln Leu Cys Phe Tyr Phe Asp Glu Glu Ala Glu Gln Lys Val Met
145 150 155 160
Pro Leu Leu Pro Gly Leu Ser Ala Thr Arg Trp His Pro Leu Phe Ala
165 170 175
Asp Val Asn Val Ala Gly Thr Ser Lys Ala Thr Gly Leu Ser Leu Phe
180 185 190
Ala Asp Tyr Tyr Arg Val Lys Val Ser Glu Ile Met Ala Cys Gly Asp
195 200 205
Gly Gly Asn Asp Ile Pro Met Leu Lys Ala Ala Gly Ile Gly Val Ala
210 215 220
Met Gly Asn Ala Ser Glu Lys Val Gln Ser Val Ala Asp Phe Val Thr
225 230 235 240
Asp Thr Val Asp Asn Asp Gly Leu Tyr Lys Ala Leu Lys His Phe Gly
245 250 255
Val Ile

Claims (4)

1.一种6-磷酸氨基葡萄糖磷酸酶突变体，其特征在于，所述突变体的序列如SEQ IDNO.10、SEQ ID NO.12、SEQ ID NO.14、SEQ ID NO.16、SEQ ID NO.18、SEQ ID NO.20、SEQ IDNO.22、SEQ ID NO.24、SEQ ID NO.26、SEQ ID NO.28、SEQ ID NO.30、SEQ ID NO.32、SEQ IDNO.34、SEQ ID NO.36、SEQ ID NO.38、SEQ ID NO.40、SEQ ID NO.42、SEQ ID NO.44、SEQ IDNO.46、SEQ ID NO.48、SEQ ID NO.50、SEQ ID NO.52、SEQ ID NO.54、SEQ ID NO.56、SEQ IDNO.58和SEQ ID NO.60所示。
2.一种多核苷酸，所述多核苷酸包含编码权利要求1所述的突变体的核苷酸序列。
3.一种宿主细胞，所述宿主细胞包含权利要求2所述的多核苷酸。
4.权利要求1所述突变体，权利要求2所述多核苷酸，权利要求3所述宿主细胞在制备氨基葡萄糖中的应用。
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CN108410839A (zh) * 2018-01-24 2018-08-17 北京理工大学 一种热稳定性提高的β-葡萄糖醛酸苷酶突变体

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