CN114395018A - CifA和CifB突变蛋白用于调控胞质不相容性的用途 - Google Patents

CifA和CifB突变蛋白用于调控胞质不相容性的用途 Download PDF

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CN114395018A
CN114395018A CN202111665809.2A CN202111665809A CN114395018A CN 114395018 A CN114395018 A CN 114395018A CN 202111665809 A CN202111665809 A CN 202111665809A CN 114395018 A CN114395018 A CN 114395018A
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王泽方
肖云杰
王镐锋
王炜
陈侠
杨海涛
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Tianjin University
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Abstract

本发明公开了一种人工设计的胞质不相容性因子(CI factor，Cif)包括CifA(CidA,CinA，CndA)和CifB(CidA，CidB，CndB)的突变蛋白用于调控胞质不相容性的用途。本发明的CifA或CifB突变蛋白能够使得本来兼容的A、B因子失去相互作用，使得来自不同沃尔巴克氏体(Wolbachia)菌株的本不相容的胞质不相容性因子CifA与CifB产生相互作用。本发明的突变蛋白通过实验证实具有野生型胞质不相容性因子的功能，能够通过结合B因子从而消除B因子诱导的酵母生长抑制，实现酵母拯救的能力；同时对B因子的突变也未破坏其诱导酵母毒性的能力。另外本发明的突变蛋白能够在果蝇水平实现调控胞质不相容性现象，说明本发明的CifA和CifB突变蛋白能够应用于调控胞质不相容性，实现胞质不相容性的可控操纵。

Description

CifA和CifB突变蛋白用于调控胞质不相容性的用途
技术领域
本发明属于蛋白的基因工程领域，具体的涉及一种CifA和CifB突变蛋白的用途。
背景技术
蚊媒疾病一直以来危害人类的生命安全，每年有数百万人感染登革热、基孔肯雅热和寨卡病毒，已成为全球公共卫生安全问题。蚊虫作为重要的媒介生物，是蚊媒传染病控制的重点。随着生物防治蚊虫技术的发展，基于昆虫共生菌沃尔巴克氏体(Wolbachia)的蚊媒和蚊媒病控制逐渐成为研究及应用热点。沃尔巴克氏体能够引起胞质不相容性(CI)现象，即沃尔巴克氏体感染的雄性与未感染的雌性交配，产生的后代无法存活；此外，携带不同Wolbachia菌株的蚊子交配后代也无法正常发育。CI是一种基因驱动机制，能够影响种群结构并引起生殖隔离。2017年，胞质不相容性因子(CI factor,Cif)被发现，这类因子由两个蛋白组成，分别被称为胞质不相容性因子CifA(包括CinA,CidA)和CifB(包括CinB,CidB)。它们能够不依赖其它Wolbachia因子介导胞质不相容性。研究发现A、B因子在体外能够发生直接相互作用，并且酵母生长实验及转基因果蝇实验证实CifA和CifB之间的相互作用对于拯救CI至关重要。同时，研究发现感染不同Wolbachia菌株的蚊虫互不兼容，即由不同的Wolbachia菌株诱导的CI表型出多样性。例如来自wMel的CidA不能与来自wPip的CidB发生相互作用，也即不能表现出CI拯救的现象。
目前，尚未有一种将胞质不相容性因子CifA、CifB进行突变，从而实现CifA与CifB解除相互作用，同时使来自不同菌株的CifA与CifB产生相互作用的报道。尽管胞质不相容性因子被发现，但对于调控其相互作用，进而应用其用于蚊媒疾病和农业害虫防控等方面仍然存在不足。
发明内容
1.本发明的目的是克服现有技术的不足，提供一种CifA和CifB突变蛋白用于调控胞质不相容性的用途。
本发明的技术方案概述如下：
1.CifA和CifB突变蛋白用于调控胞质不相容性的用途。
2.本发明中的CifA和CifB突变蛋白，包括但不限于列举出的突变蛋白，其中突变蛋白CidAST的氨基酸序列是SEQ ID NO.2所示；突变蛋白CidBST的氨基酸序列是SEQ IDNO.4所示；突变蛋白CinAST的氨基酸序列是SEQID NO.6所示；突变蛋白CinBST的氨基酸序列是SEQ ID NO.8所示。
本发明的优点：
本发明设计的CifA和CifB突变蛋白能够用于调控胞质不相容性。本发明的CifA和CifB的突变蛋白能够正确折叠，在大肠杆菌系统能够大量纯化；利用酵母系统验证胞质不相容性因子突变蛋白与野生型相比仍然具有活性，A因子保持酵母拯救的能力，B因子能够诱导酵母死亡。同时CinAST失去与CinB相互作用的能力；CidAST能够与本不兼容的wPipCidB产生相互作用，并且酵母实验证明CidAST能够拯救wPip CidB诱导的酵母毒性。另外，转基因果蝇实验证明突变蛋白能够在果蝇水平调控胞质不相容性现象。本发明的CifA和CifB突变蛋白在生化、酵母、果蝇水平均能实现对胞质不相容性的可控调控，解决胞质不相容性因子在不同系统中不兼容的问题，赋予了来自不同菌株胞质不相容性因子相互兼容的范围及能力，对利用胞质不相容性防控蚊媒疾病和农业害虫具有重要的意义，对实现胞质不相容性的可控操纵具有指导作用，可应用于蚊媒防控、农业害虫防控等领域。
附图说明
图1为wPip CidA-CidBND1-ND2及CinA-CinB蛋白复合物结构图。
图2为CidAST蛋白凝胶色谱层析检测图及聚丙烯酰胺凝胶电泳检测图。
图3为CinAST蛋白凝胶色谱层析检测图及聚丙烯酰胺凝胶电泳检测图。
图4为CidAST与wPip CidB互作分析图。
图5为CinAST与wPip CinB互作分析图。
图6为CinBST与wPip CinA互作分析图。
图7为CidBST与wPip CidA互作分析图。
图8为CidAST在酵母中拯救wPip CidB诱导酵母毒性分析图。
图9为CinAST、CinBST酵母表型分析图。
图10为CidBST酵母表型分析图。
图11为CinAST用于转基因果蝇孵化率分析图。
具体实施方式
下面通过具体实施例对本发明作进一步的说明。
PET-22b，pRS425和pRS416为市售，大肠杆菌BL21(DE3)和酿酒酵母BY4741为市售。
实验材料：
(1)LB培养基：配制每1000mL培养基，在1000mL的一次蒸馏水中加入蛋白胨10g，酵母粉5g，NaCl 10g，溶解后，121℃、0.1MPa灭菌20min。配置固体LB培养基时向培养基内补加1.5％的琼脂粉。
(2)LB固体培养基：准确称取蛋白胨5g，酵母粉2.5g，NaCl 5g，琼脂糖A7.5g，加入单蒸水定容至500mL，0.1MPa，121℃灭菌20min，待冷却至约50℃时，加入千分之一体积的氨苄青霉素，倒入100mm平板中，冷却后放置于4℃备用。
(3)YPD培养基：配制每1000mL培养基，在1000mL的一次蒸馏水中加入蛋白胨20g，酵母粉10g，NaCl 8g，葡萄糖20g。
(4)Ampicillin青霉素(100mg/mL)：准确称取20mg氨苄青霉素固体粉末，加入到20mL灭菌的去离子水中，轻摇使其充分溶解，使用移液器分装，每管200微升，冻存到-20℃冰箱备用。
(5)异丙基硫代-β-D-半乳糖苷(IPTG)：在无菌环境下，将准确称取的10gIPTG粉末溶解与42mL灭菌的双蒸水中，充分混匀，500μL每管进行分装，保存于-20℃冰箱备用。
(6)50×TAE电泳缓冲液：准确称取Tris 242g，使用量筒量取100mL的0.5mM的EDTA溶液，加入57.1mL的冰醋酸，调节pH至8.0，置于室温备用。
(7)SDS-PAGE(聚丙烯酰胺凝胶电泳)
10％APS溶液：准确称取1g APS固体粉末，加入灭菌水10mL使其充分溶解，使用1.5mL EP管分装，每管200微升，冻存于-20℃冰箱备用。
10％SDS溶液:准确称取1g SDS固体粉末，加入灭菌水10mL使其充分溶解，使用1.5mL EP管分装，每管500微升，放置于-20℃冰箱备用。
30％丙烯酰胺溶液(100mL)：将30g丙烯酰胺与0.8g的甲叉双丙烯酰胺溶解在100mL的双蒸水中，溶解后放置于4℃棕色瓶内。
1.5M Tris-HCl pH＝8.8缓冲液(1L)：称取181.71g Tris溶解在800mL纯水中，调pH至8.8，定容至1L，室温保存。
0.5M Tris-HCl pH＝6.8缓冲液(500mL)：称取60.57g Tris溶解在400mL纯水中，调pH至6.8，定容至500mL，室温保存。
表1 12％SDS-PAGE分离胶
Table 2-3 Separation Gel recipe of SDS-PAGE(12％)
Figure BDA0003451086290000041
表2 SDS-PAGE浓缩胶
Table 2-4 Condension Gel recipe of SDS-PAGE
Figure BDA0003451086290000042
(8)5×Tris-甘氨酸电泳缓冲液(5L)：称取75.5g Tris，470g甘氨酸，25g SDS，加纯水溶解，定容至5L。
(9)6×蛋白上样缓冲液：0.35M pH＝6.8Tris-HCl，10.28％W/V SDS，36％甘油，5％β-巯基乙醇，0.012g/mL溴酚蓝，1mL每管分装，-20℃保存备用。
(10)酵母表型实验所用试剂
1)10X TE buffer(100mM Tris，pH 7.5,10mM EDTA)：称取1.21g Tris，0.34gEDTA-Na2，70mL水溶解，Hcl调pH至7.5,定容至100mL。
2)10X LTE buffer:称取10.2g二水醋酸锂，使用上述10X TE buffer溶解，定容至100mL，抽滤。
3)40％(w/v)PEG3350:40g PEG3350,使用1X LTE buffer溶解，定容至100mL。
4)20％(w/v)葡萄糖：称取40g葡萄糖，双蒸水定容至200mL，0.1MPa，115℃灭菌20min。
5)20％(w/v)半乳糖：称取40g半乳糖，双蒸水定容至200mL，0.1MPa，115℃灭菌20min。
6)50X(SD-URA)混合氨基酸：称取0.1g精氨酸，0.05g组氨酸，0.3g异亮氨酸，0.3g亮氨酸，0.2g赖氨酸，0.05g蛋氨酸，0.3g苯丙氨酸，0.25g苏氨酸，0.2g色氨酸，加100mL双蒸水,抽滤灭菌，4℃保存备用。
7)50X(SD-URA-LEU)混合氨基酸：称取0.1g精氨酸，0.05g组氨酸，0.3g异亮氨酸，0.2g赖氨酸，0.05g蛋氨酸，0.3g苯丙氨酸，0.25g苏氨酸，0.2g色氨酸，加100mL双蒸水,抽滤灭菌，4℃保存备用。
8)1％adenine：称取1gadenine，溶于0.1M 100mL的NaOH中，抽滤除菌。
9)2％Glu-SD-URA：称取1.34g YNB，加176mL双蒸水，121℃灭菌20min，冷却后加入20％葡萄糖20mL,50X(-URA)混合氨基酸4mL，1％adenine 0.4mL。固体培养基向其中加入5g琼脂。
10)2％Gal-SD-URA：称取1.34g YNB，加176mL双蒸水，121℃灭菌20min，冷却后加入20％葡萄糖20mL,50X(-URA)混合氨基酸4mL，1％adenine 0.4mL。固体培养基向其中加入5g琼脂。
11)2％Glu-SD-URA-LEU：称取1.34g YNB，加176mL双蒸水，121℃灭菌20min，冷却后加入20％葡萄糖20mL,50X(-URA-LEU)混合氨基酸4mL，1％adenine 0.4mL。固体培养基向其中加入5g琼脂。
12)2％Gal-SD-URA-LEU：称取1.34g YNB，加176mL双蒸水，121℃灭菌20min，冷却后加入20％葡萄糖20mL,50X(-URA-LEU)混合氨基酸4mL，1％adenine 0.4mL。固体培养基向其中加入5g琼脂。
下面结合实施例对本发明做进一步说明，以下所述，仅是本发明的部分较佳实施例，并非对本发明做其他形式的限制。凡是未脱离本发明的方案内容，依据本发明的技术实质对实施例所做的任何修改或等同变化，均在本发明的保护范围内。
实施例1 pET-22b-CidAST的构建
以黑腹果蝇(Drosophilamelanogaster)中SEQ ID NO.9所示的wMel CidA核苷酸序列及来自尖音库蚊(Culex pipiens)中SEQ ID NO.10所示的wPip CidA核苷酸序列为模板，以wPip CidA-CidBND1-ND2(wPip CidB核苷酸酸序列为SEQ ID NO.11所示，氨基酸序列为SEQ ID NO.12所示)及wMel CidA的结构为基础，其中A、B因子相互作用界面可以分成三个，将wPip CidA中参与CidA与CidB相互作用的氨基酸残基替换到wMel CidA上，设计出CidAST序列，如SEQ ID NO.1所示。以上述人工合成的SEQ ID NO.1为模板，以F1为正向引物，以R1为反向引物，经过PCR，得到的含有CidAST的基因序列，经酶切及T4 DNA连接酶连接，将突变蛋白CidAST基因构建到PET-22b载体上，得到PET-22b-CidAST质粒。具体步骤如下：
设计并通过基因合成获得CidAST基因序列后，首先设计引物进行PCR，正向引物和反向引物分别命名为F1和R1，扩增目的基因，PCR体系如表3所示。
表3 PCR体系
Figure BDA0003451086290000061
PCR反应条件为：95℃预变性3min，95℃变性30s，50-60℃设置温度梯度进行退火30s，72℃延伸2min，设置33个循环，最后再延伸10min反应结束。在反应结束后，使用1％琼脂糖凝胶电泳进行电泳，然后使用胶回收试剂盒进行胶回收，得到PCR产物。
将上述得到的PCR产物以及pET-22b(+)等空载在37℃条件下进行双酶切(酶切体系见表4)，反应时间30min，使用酶为NdeI和XhoI。酶切完成后使用1％琼脂糖凝胶电泳进行电泳，然后使用胶回收试剂盒进行胶回收，得到含粘性末端的基因及pET-22b(+)载体。然后使用T4 DNA连接酶在22℃下进行连接(连接体系见表5)，连接反应时间1h。
表4双酶切体系
Figure BDA0003451086290000071
表5连接体系
Figure BDA0003451086290000072
连接反应完成后，将20μL的连接产物加入到100μL的E.coli DH5α感受态中，冰上静置30min，接着，42℃水浴热激90s，之后快速插入冰中，静置5min，加入500μL LB液体培养基，37℃摇床复苏45min。将复苏后的菌体离心，3000rpm，4min，吸弃上清，留约100μL上清液将菌体轻柔混匀，使用涂布棒均匀涂布在含100μg/mL氨苄青霉素的LB固体培养基上，放置在37℃培养箱中过夜培养。第二天挑取单菌落到含100μg/mL氨苄青霉素的5mL的LB中，37℃摇床震荡培养10-12h。使用天根质粒小提试剂盒提取质粒，接着将提取的质粒进行双酶切验证，双酶切验证体系见表6，将该反应体系置于37℃水浴反应30min，反应完成后进行琼脂糖凝胶电泳，将酶切验证正确的质粒送测序公司进行测序验证。
表6双酶切验证体系
Figure BDA0003451086290000073
实施例2 pRS425-CidAST的构建
选择载体为pRS425，酶切位点为BamHI和SalI，设计引物进行PCR，引物分别命名为F2和R2，PCR体系如表3所示。将上述实施例1构建的pET-22b-CidAST为模板，PCR可得到酶切位点为BamHI和SalI的第二种含有突变蛋白CidAST基因的序列。
用限制性内切酶BamHI和SalI对上述PCR产物和载体pRS425分别进行双酶切，使目的基因片段和载体出现粘性末端，具体酶切体系如表4所示。
将有粘性末端的目的基因片段和载体pRS425使用T4DNA连接酶进行连接，连接体系如上表5所示。
再经过转化及双酶切验证，将得到的阳性结果进行测序验证。最终得到pRS425-CidAST构建成功质粒。
实施例3 pRS416-CidBST的构建
以黑腹果蝇(Drosophila melanogaster)中SEQ ID NO.17所示的wMel CidB核苷酸序列及来自尖音库蚊(Culex pipiens)中SEQ ID NO.11所示的wPip CidB核苷酸序列为模板，以wPip CidA-CidBND1-ND2的结构为基础，将wPip CidB中参与CidA与CidB相互作用的氨基酸残基替换到wMel CidB上，设计出CidBST序列，如SEQ ID NO.3所示。以上述人工合成的SEQ ID NO.3为模板，以F3为正向引物，以R3为反向引物，经过PCR，得到的含有CidBST的基因序列，经酶切及T4 DNA连接酶连接，将突变蛋白CidBST基因构建到PET-22b载体上，得到PET-22b-CidBST质粒；以PET-22b-CidBST质粒为模板，以F4为正向引物，以R4为反向引物，进行PCR，得到第二种含有突变蛋白CidBST基因的序列，经酶切及T4 DNA连接酶连接，将突变蛋白CidBST基因构建到pRS416载体上，得到pRS416-CidBST质粒。具体步骤如下：
选择载体为pRS416，酶切位点为BamHI和SalI，设计引物进行PCR，引物分别命名为F4和R4，PCR体系如表3所示。将人工合成的wPip CidBST基因为模板，PCR可得到酶切位点为BamHI和SalI的含有蛋白CidB基因的序列。
用限制性内切酶BamHI和SalI对上述PCR产物和载体pRS416分别进行双酶切，使目的基因片段和载体出现粘性末端，具体酶切体系如表4所示。
将有粘性末端的目的基因片段和载体pRS416使用T4DNA连接酶进行连接，连接体系如上表5所示。
再经过转化及双酶切验证，将得到的阳性结果进行测序验证。最终得到pRS416-CidBST构建成功质粒。
实施例4 pRS416-CidB的构建
选择载体为pRS416，酶切位点为BamHI和SalI，设计引物进行PCR，引物分别命名为F9和R9，PCR体系如表3所示。将人工合成的wPip CidB基因为模板，PCR可得到酶切位点为BamHI和SalI的含有蛋白CidB基因的序列。
用限制性内切酶BamHI和SalI对上述PCR产物和载体pRS416分别进行双酶切，使目的基因片段和载体出现粘性末端，具体酶切体系如表4所示。
将有粘性末端的目的基因片段和载体pRS416使用T4DNA连接酶进行连接，连接体系如上表5所示。
再经过转化及双酶切验证，将得到的阳性结果进行测序验证。最终得到pRS416-CidB构建成功质粒。
实施例5突变蛋白CidAST表达及纯化
将含有pET-22b-CidAST的重组质粒转化到E.coli BL21(DE3)中，过夜培养，挑取单菌落到5mL含100μg/mL的Ampicillin的LB培养基中，37℃摇床震荡培养6-8h；随后，将菌液转入1L含100μg/mL氨苄青霉素的LB培养基中，37℃摇床震荡培养6-8h，菌液培养至OD600为0.6左右，降温至16℃，加入IPTG至终浓度400μM，诱导培养16-18h。随后离心(4000rpm，20min)收集菌体，将收集的菌体使用悬菌buffer(20mM Tris-HCl,pH 7.5,0.3M NaCl,10μg/ml DNase1,10μg/ml RNaseA and 10％glycerol)重悬，每升菌体约使用15mL悬菌buffer，之后使用高压破菌仪进行破菌。破菌完成后，将破菌产物使用冷冻高速离心机离心(18000rpm，4℃，45min)，收集上清，加入到预先使用悬菌buffer平衡过的镍亲和层析填料，4℃孵育1h。孵育完成后，在4℃环境下使用洗杂buffer(20mM Tris-HCl,pH 7.5,0.3MNaCl,10％glycerol and 20mM imidazole)洗杂，约使用20倍填料体积的洗杂buffer分多次对该体系进行冲洗，以便将非特异性结合到Ni柱上的杂蛋白清洗干净。在清洗干净后，使用约30mL洗脱液(20mM Tris-HCl,pH 7.5,0.3M NaCl,10％glycerol and 300mMimidazole)将结合在Ni柱上的目的蛋白洗脱。收集洗脱液，使用30kDa浓缩管进行浓缩，并使用A液(20mM Tris-HCl,5mM DTT,pH 7.5)进行换液，用以将蛋白溶液进行降盐，降至NaCl终浓度50mM左右，继续离心浓缩。
将浓缩后蛋白使用AKTApure进行纯化，使用阳离子交换柱(Hi Trap SP 5ml,GE)。缓冲液分别使用A液和B液(20mM Tris-HCl,1M NaCl,5mM DTT,pH 7.5)。洗脱梯度按50mM-600mM NaCl，洗脱总体积80mL，收集流出组分并使用12％的SDS-PAGE检测各个组分，根据分子量大小收集目的蛋白。接着，将收集的较纯的蛋白使用4℃离心机浓缩，转速3400rpm，约40min，浓缩到体积500μL以下,使用AKTApure进行纯化。使用预装柱(Superdex 20010/300GL,GE)纯化，分子筛buffer为20mM Tris-HCl,150mM NaCl，5mM DTT，pH 7.5，程序设置为流速0.5mL/min，收集流出组分并使用12％的SDS-PAGE检测各个组分，根据分子量大小收集目的蛋白。
实施例6突变蛋白CidAST与wPip CidB蛋白pull down
为验证胞质不相容性因子A蛋白突变后与B因子间是否存在相互作用，使用6x HisPull-down实验方案进行。wPip CidB氨基酸序列为SEQ ID NO.6所示。具体6x His Pull-down实验方案如下：
将带His-tag标签的A蛋白与B蛋白混合，并保证B蛋白过量。蛋白混合均匀后，向体系中加入Ni介质30μL，同时向体系中加入结合buffer(20mM Tris-HCl pH 7.5,200mMNaCl,0.01％Tween-20,5％Glycerol)至终体积为600μL，置于4℃使用滚轴混悬器孵育1h。孵育完成后，将混合体系加入到1.5mL离心式微量蛋白纯化柱内，通过离心(700g，1min)的方法进行洗杂，洗杂buffer(20mM Tris-HCl pH 7.5,200mM NaCl,0.01％Tween-20,5％Glycerol,50mM imidazole)。使用洗杂buffer反复洗8次，使得不能结合Ni介质的蛋白清洗干净。洗杂完成后，使用洗脱液(20mM Tris-HCl pH 7.5,200mM NaCl,0.01％Tween-20,5％Glycerol,300mM imidazole)进行洗脱操作，每管分两次进行洗脱，每次加入50μL洗脱液。洗脱完成后，向每管洗脱液中加入6x的蛋白loading20μL，然后将样品置于金属浴进行加热，100℃加热10min，加热完成后，使用离心机离心，12000rpm，5min。然后，使用12％的SDS-PAGE检测。
实施例7突变蛋白CidAST用于酵母表型分析
将突变蛋白CifAST用于酵母表型分析，用于验证突变后是否能够结合wPip CidB蛋白。具体实施方案如下：
一、醋酸锂转化
首先将pRS416和三种wPipCidB进行单独转化。
步骤：
1)使用YPD小管过夜培养Yeast BY4741菌株，共培养三支小管；
2)使用4个无菌EP管，分别吸取500μl菌液离心，5000xg，2min；使用1mL无菌水重悬；离心，5000xg，2min；
3)使用50μl 1x LTE buffer重悬，分别加入6.75μl 10mg/ml的ssDNA(ssDNA使用前沸水浴5min后立即置于冰上)，加入0.1-0.5μg上述质粒，加入400μl含40％的PEG-3350的1X LTE buffer，混合均匀，室温孵育30min。
4)42℃孵育20min；之后离心，5000xg，2min；使用100μL无菌水重悬；涂板，使用2％葡萄糖的SD-URA固体培养基，30℃培养三天。
二、CidB阳性克隆筛选
将上述步骤中进行的转化(pRS416空载，CidB)，每种各选取3个单克隆到新的含2％半乳糖或2％葡萄糖的SD-URA固体培养基，置于30℃培养3天，观察板子菌落生长状态，筛选出能够对Yeast产生毒性的克隆。
三、将pRS425，wPipCidA，wMel CidA，CidAST分别转化到上述筛选出的CidB阳性克隆及pRS416空载，使用2％葡萄糖的SD-URA-LEU(固)。具体步骤如下：
分别将第二步实验中筛选出的pRS416空载，CidB阳性克隆使用2％葡萄糖的SD-URA液体培养基过夜培养，每种培养3个小管。按照上述转化过程，将pRS425空载或几种CidAvariants转化到对应的CidB或pRS416空载中，使用2％葡萄糖的SD-URA-LEU固体培养基涂板，30℃培养三天。
四、酵母表型分析
将上述转化得到的转化子各选取3个单克隆到新的含2％半乳糖或2％葡萄糖的SD-URA-LEU固体培养基，置于30℃培养3天，观察板子菌落生长状态，观察CidA突变对于Yeast的拯救情况。挑选出符合预期的转化子。
接下来进行菌斑培养，使用6倍稀释法，每个转化子稀释6个菌斑，起始OD600为0.2。使用2％半乳糖的SD-URA-LEU固体培养基；葡萄糖对照组每种转化子培养一组菌斑，使用2％葡萄糖的SD-URA-LEU固体培养基。其中6倍稀释法具体步骤如下：
1.在YPD培养基中过夜培养上述筛选的转化子。
2.测定每种培养物的OD600，首先在培养基中稀释10倍，并测量OD600。
3.计算稀释到OD600为0.2时需要的培养基中的酵母体积，每个样品制备1mL OD600为0.2的菌液。
4.将125μL无菌ddH2O添加96孔板的B列至F列。
5.将125μL每种培养物添加到96孔板的第1列(即，菌株1进入1A，菌株2进入2A等)
6.将25μL菌液从A孔转移至B孔；混合将25μL从B转移到C；混合重复第F列。
7.使用多通道移液器混合每个样品，并将2μL点在平板上，从F开始，向A移动(密度从最低到最高)
8.让所有斑点干燥；转移到30℃的培养箱。
9.每天监测生长情况，三天后拍照。
实施例8突变蛋白CinAST(I-III)用于转基因果蝇孵化率分析
将突变蛋白CinAST(I-III)用于转基因果蝇孵化率分析，用于验证突变后是是否影响CI表型，影响果蝇孵化率，其中CinAST(I-III)氨基酸序列如SEQ ID NO.34所示，核苷酸序列如SEQ ID NO.35所示。具体实施方案如下：
通过Gal4 UAS系统在果蝇中实现CI因子转基因的控制表达，通过公司Genscript合成cinAST(I-III)的ORF，并使用Not I和BamH I限制性位点将其亚克隆到pUASp-attB载体中。研究中使用的所有转基因果蝇均为9744背景，并在其第三条染色体上整合了转基因。研究中使用的所有转基因果蝇均保持为纯合系。
果蝇种群在18℃的标准玉米粉固体培养基上以12小时：12小时的光照：黑暗循环进行饲养。孵化率分析中使用的亲代果蝇是通过将MTD-Gal4雌蝇与非转基因雄蝇杂交产生的。收集所需基因型和性别的果蝇，单独保存，并在室温下饲养2-4天。将一只雄性和一只雌性转移到一个倒置的100mL聚丙烯罐中，底部穿孔通气，开口由涂有酵母膏的苹果汁盘覆盖。成对果蝇至于25℃的孵化器里，将苹果汁盘放在底部过夜(～16小时，实际温度在25℃-26.5℃之间波动)。第二天早上，用新盘子替换旧盘子，让果蝇再产卵24小时。然后收集亲代苍蝇，并在-20℃冷冻，放置在苹果汁盘上的胚胎在25℃时孵化最多48小时。在显微镜下对胚胎和未孵化胚胎的总数进行计数。两个板块的数据汇集在一起。不包括来自总产卵量少于15枚的交配雌性的数据。
突变蛋白CidAST核苷酸序列SEQ ID NO.1(人工合成)
ATGCCAATCGAAACCAAGAAACAAGCCGAGGTGCTGAAGAAGCTGCAAGATGTGATCAAGCATACCGATCGCGATATCGCGGCCGGCCGTAAACTGGCCATTAAACGCTGGGTGGAGACGTATATTGAATATATCAAATACTTTAAGGACGACAAGCTGGAGTTTCTCTACAACGTGTTCCGTGACGAGGGCTGTTGGCTGGGCACCCGCCTCAACAACACCGTGCTCGGTCAGAAACTCACGGAGGAGAAAATCGGCGAGATCGATAACCCGCTGCGCCGTTACGGCATGGCCAGTCGTTACTGCATCACCGGCAAAATCCACCCGCTGTTCCAAAAACGCTTCGAGAGTTACCGCAATAAGTTCCCACCGGGTGCCTTCGACGGCAAGACCGAGACGGAATTCGGCAAGTACGTTCGTAACAGTCTGCTGGATAGCATCAAGCGCAAAGGCCCGGTTTTCGACTTTTGGATCGACCGCGAGAGCGGCGAGCTGAAGAAATACGATGCCGTGGAGGGCTTTGACAGCACGGTGAAGCTGAAGTGGAGCGAGGGTGTTGAGTACTTCTACAACCAGCTGGAAGAGAAGGATAAGGAGAAGAAACTGACCGAAGCCATCGTGGCGCTGAGTCGCCCACAGAGTGTTAAACGCGATGCGCCGATTCTGGATTTCTGCGTGCGCAATATCGGCGACAAAGATACGCTGCTGCAGAAACTGCTGCAGAAGGACAAAGGCGTGTATTTTCTGCTCGCCGAGCTCATCGAGAGCTGTTTCTTTGACACCGTTCACGATCTGGTTCAGTGCTGGTGCTACAAAGGTGTTAGCGCGGGTGGCGATTGCAGCGACAAAATCTTCAGCCAGCGCGACTACGAGCTGTTTCTGAGCAGTCTGAGCGATGTGATGCTGAAGAATCCGGAACTCAGCGTGCAAGCCCGTAGTCTGATCATGGAGATTTGGAAGTGCGAACGCTTCGCGGAATACCGCGAGACGAGTGTGAACACGAGCAACTACACCGTGCCGATCAAAAGCGTGCTGGGCGAACTGATCATCAACTGGAAACGCGAAGACGTGTGTAAACCGGATCGCGAGATCGAAAAAGAAGAAATTCTGGATATGATCAGCTTCGCGAAGGGCTGCTTCCCGGAGAAATTCGATCTGTTCAAAGAAGTTATGATCCGCAATCTCCGTCTGTGCGGTCGCGAGGGCAAACGCAAGGGTGTGGATTACGGTAAGTTCGCGGAGGAACTCTTCCTCCAGCTGGAAAAGGTTACGCTGCCGAGCGTGGGCGATGGTCCGTGGAATAATCTGCGCAGCCAGAGCAAAGTGAGTCTGCCGCTGGATGGCAGTGGCGACGGTCCACAAAGTGAATTCGAAGCCCCGAGTGTGAGCGGCATTAGCGGCAGCCATAAAAAACGCCGCATC
突变蛋白CidAST氨基酸序列SEQ ID NO.2(人工合成)
MPIETKKQAEVLKKLQDVIKHTDRDIAAGRKLAIKRWVETYIEYIKYFKDDKLEFLYNVFRDEGCWLGTRLNNTVLGQKLTEEKIGEIDNPLRRYGMASRYCITGKIHPLFQKRFESYRNKFPPGAFDGKTETEFGKYVRNSLLDSIKRKGPVFDFWIDRESGELKKYDAVEGFDSTVKLKWSEGVEYFYNQLEEKDKEKKLTEAIVALSRPQSVKRDAPILDFCVRNIGDKDTLLQKLLQKDKGVYFLLAELIESCFFDTVHDLVQCWCYKGVSAGGDCSDKIFSQRDYELFLSSLSDVMLKNPELSVQARSLIMEIWKCERFAEYRETSVNTSNYTVPIKSVLGELIINWKREDVCKPDREIEKEEILDMISFAKGCFPEKFDLFKEVMIRNLRLCGREGKRKGVDYGKFAEELFLQLEKVTLPSVGDGPWNNLRSQSKVSLPLDGSGDGPQSEFEAPSVSGISGSHKKRRI
突变蛋白CidBST核苷酸序列SEQ ID NO.3(人工合成)
ATGTCTAACGGTGACGGTCTGATCCGTTCTCTGGTTGACGGTGACCTGGAAGGTTTCCGTCAGGGTTTCGAATCTTTCCTGGACCAGTGCCCGTCTTTCCTGTACCACGTTTCTGCGGGTCGTTTCCTGCCGGTTTTCTTCTTCTCTATGTTCTCTACCGCGCACGACGCGAACATCCTGAACGCGAACGAACGTGTTTACTTCCGTTTCGACAACCACGGTGTTAACCCGCGTAACGGTGAAAACCGTAACACCGCGAACCTGAAAGTTGCGGTTTACCGTGACGGTCAGCAGGTTGTTCGTTGCTACTCTATCTCTGACCGTCCGAACTCTGACGGTCTGCGTTTCTCTACCCGTGAACGTAACGCGCTGGTTCAGGAAATCCGTCGTCAGAACCCGAACCTGCGTGAAGAAGACCTGAACTTCGAACAGTACAAAGTTTGCATGCACGGTAAAGGTAAATCTCAGGGTGAAGCGATCGCGACCGTTTTCGAAGTTATCCGTGAGAAGGACCGTCAGGGTCGTGACAAATTCGCGAAATACTCTGCGTCTGAAGTTCACTTCCTGCGTCAGCTGTTCCGTAACCACCGTCTGACCATCAAAGAAATCGAAGGTCGTCAGCTGAACCAGAACCAGCTGCGTCAGCTGGGTCGTTCTGTTAACTTCACCCGTGTTGAACCGGGTCAGCAGGGTATCGACAACTTCATGGAAATGCTGGCGTCTAACCAGAAAATTAACGTTGAGGACTCTCTGCGTGGTGACATCCTGGAATACGTTACCCGTACCTACAACAACTACCGTGCGCAGATCGAAAACAACATCGAAGGTCGTTCTCAGAAATTCGAATCTCACGGTTTCCTGCTGGGTTTCCTGGCGAACTTCTCTCACCGTTACACCATCGGTGTTGACCTGGACCTGTCTCCGCGTAACTCTCACGTTGCGTTCCTGGTTCGTCACCAGGTTGAACGTGAAAACATCCCGATCGTTATCAACCTGGCGACCCGTGCGCCGCCGTACATCGCGCTGAACCGTGCGCGTTCTCACGCGAAACGTCTGCACGTTTTCTCTTTCATCCCGATCCACACCGAATCTCGTAACACCGTTTGCGTTGGTCTGAACTTCAACCTGAACCTGGACCCGTTCTCTGTTGACACCGTTGGTCTGCAGCAGGACCGTTTCCCGCTGGTTCAGCGTCTGTTCGAATGCCTGGAAAACGAAGGTATCCGTGAAAACATCCGTGACTTCCTGCTGCACCACCTGCCGGCGGAAATCCCGCGTAACGCGGAAAACTACGACCGTATCTTCGACTGCATCACCGGTTTCGCGTTCGGTAACTCTGCGTTCGACGAACACCCGCTGGTTAATGAAGAAGAAGTTCAGGCGCCGATCACCAAATACATCTTCCGTCACGGTGACGAAGGTCTGCGTTGCCTGACCATGGTTTTCCACGCGGAAGGTTCTGACATCGTTATCCTGCACATCCGTGCGCACGACGCGCAGCAGCAGGGTGCGATCAACCTGCAGACCCTGAACGTTAACGGTAACGACGTTCACGTTTGGGAAGTTTCTTGCACCCTGAACAACCAGCTGGAACTGGACATCGACCTGCCGAACGACCTGGGTCTGTACCACGACTACCAGAACAACAACGCGAACAACTTCCTGGCGGGTGACCTGGTTCAGGTTCCGAACACCGAAAACGTTCACAACACCCTGAACCAGGTTGTTAACGACGGTTGGAAGAATATCGCGCAGCACCGTGGTCTGTTCCAGGAAATCTCTGGTGCGCTGATGCCGCTGGTTGACACCATCAACGTTAACTCTGAAGACAAATTCCGTTCTATCCTGCACGGTACCTTCTACGCGTCTGACAACCCGTACAAAGTTCTGGCGATGTACAAAGTTGGTCAGACCTACTCTCTGAAACGTGGTCAGGAAGAAGAAGGTGAACGTGTTATCCTGACCCGTATCACCGAACAGCGTCTGGACCTGCTGCTGCTGCGTCAGCCGCGTGAAAACGACCTGGACACCCACCCGATCGGTTACGTTCTGCGTCTGGCGAACAACGCGGAAGAAGTTGGTCAGCAGCAGAACGACGCGCGTCAGGAAATCGGTCGTCTGAAGAAGCAGCACCGTGGTTTCATCCCGATCACCTCTGGTAACGAAGTTGTTCTGTTCCCGATCGTTTTCAACCGTGACGCGCACGAAGCGGGTAACCTGATCCTGTTCCCGGAAGGTATCGGTCGTGAAGAACACGTTCACCGTCTGGACCGTCACGTTCGTTCTTCTCGTCCGGGTGGTCTGGTTGGTCCGGAATCTGTTATCGACGAAAACCCGCCGGAAGGTCTGCTGTCTGACCAGACCCGTGAAAACTTCCGTCGTTTCTACGAAGAGAAGGCGCCGGGTCAGAACTCTATCTTCCTGCTGGACATCGGTGACAACCTGCACGTTCCGTTCTCTTACCTGCAGGGTACCCGTGCGCAGGTTATCGAAACCCTGAAATCTCGTATCCGTGGTGGTGGTACCCCGACCGCGCAGGGTATCCTGCAGCAGATCAACGCGATCCTGCGTCGTAACAACGCGCGTGAAATCGAAGACGTTCACGACCTGCTGGCGCTGGACTTCGCGACCGACAACCAGAACTACCGTTACTGGCTGCAGACCCACGACATGTTCTTCGCGGCGCGTCAGTACACCTTCCTGGACAACCAGTCTCACTCTACCAACGACCACTACGGTTTCGAAATCACCTCTGTTGGTGTTGACGGTAACCAGAACGACCCGACCGGTCGTGGTCTGCTGTCTTCTCACATCACCAACTTCAAACAGAAAGTTGACTCTGGTGAGAAGGACCGTCTGATCGCGATCATCAACGTTGGTAACCGTCACTGGGTTACCCTGGTTATCGTTCACCAGAACGGTAACTACTACGGTTACTACGCGGACTCTCTGGGTCCGGACTCTGGTATCGACAACAACATCCGTGGTGCGCTGCGTGAATGCGACATCAACGACGACAACGTTCACAACATCTCTGTTCACCAGCAGACCGACGGTCACAACTGCGGTATCTGGGTTTACGAAAACGCGCGTGACATCAACCAGGCGATCGACCAGGCGCTGCAGGGTAACAACAACTTCGGTGAGAAGGGTGAAGGTATCATCGGTTACATCCGTGGTCTGCTGTCTGCGGGTATCGGTAACGACACCCGTCAGCCGCGTCGTAACGAACAGTACTTCGAAGACCGTCGTCGTGACATCTCTCAGCTGCTGCAGAACGACCCGAACCTGCCGTCTCGTCGTTCTGACCTGATCCAGGCGCACCCGGGTATCCAGCACGAAATCGACCCGCTGCTGCTGCAGTTCCTGGGTCTGCAGTACCCGCAGCGTGGTGGTGGTGGTGCGCTGCAGCTGGGTGGTGAACGTGTTATCTCTATCGACTTCGGTAACCCGCAGTCTGCGCTGGACAAAATCGACGGTGTTTCTCGTGTTTACAACCACTCTAACTCTCGTGGTTCTCGT
突变蛋白CidBST氨基酸序列SEQ ID NO.4(人工合成)
MSNGDGLIRSLVDGDLEGFRQGFESFLDQCPSFLYHVSAGRFLPVFFFSMFSTAHDANILNANERVYFRFDNHGVNPRNGENRNTANLKVAVYRDGQQVVRCYSISDRPNSDGLRFSTRERNALVQEIRRQNPNLREEDLNFEQYKVCMHGKGKSQGEAIATVFEVIREKDRQGRDKFAKYSASEVHFLRQLFRNHRLTIKEIEGRQLNQNQLRQLGRSVNFTRVEPGQQGIDNFMEMLASNQKINVEDSLRGDILEYVTRTYNNYRAQIENNIEGRSQKFESHGFLLGFLANFSHRYTIGVDLDLSPRNSHVAFLVRHQVERENIPIVINLATRAPPYIALNRARSHAKRLHVFSFIPIHTESRNTVCVGLNFNLNLDPFSVDTVGLQQDRFPLVQRLFECLENEGIRENIRDFLLHHLPAEIPRNAENYDRIFDCITGFAFGNSAFDEHPLVNEEEVQAPITKYIFRHGDEGLRCLTMVFHAEGSDIVILHIRAHDAQQQGAINLQTLNVNGNDVHVWEVSCTLNNQLELDIDLPNDLGLYHDYQNNNANNFLAGDLVQVPNTENVHNTLNQVVNDGWKNIAQHRGLFQEISGALMPLVDTINVNSEDKFRSILHGTFYASDNPYKVLAMYKVGQTYSLKRGQEEEGERVILTRITEQRLDLLLLRQPRENDLDTHPIGYVLRLANNAEEVGQQQNDARQEIGRLKKQHRGFIPITSGNEVVLFPIVFNRDAHEAGNLILFPEGIGREEHVHRLDRHVRSSRPGGLVGPESVIDENPPEGLLSDQTRENFRRFYEEKAPGQNSIFLLDIGDNLHVPFSYLQGTRAQVIETLKSRIRGGGTPTAQGILQQINAILRRNNAREIEDVHDLLALDFATDNQNYRYWLQTHDMFFAARQYTFLDNQSHSTNDHYGFEITSVGVDGNQNDPTGRGLLSSHITNFKQKVDSGEKDRLIAIINVGNRHWVTLVIVHQNGNYYGYYADSLGPDSGIDNNIRGALRECDINDDNVHNISVHQQTDGHNCGIWVYENARDINQAIDQALQGNNNFGEKGEGIIGYIRGLLSAGIGNDTRQPRRNEQYFEDRRRDISQLLQNDPNLPSRRSDLIQAHPGIQHEIDPLLLQFLGLQYPQRGGGGALQLGGERVISIDFGNPQSALDKIDGVSRVYNHSNSRGSR
突变蛋白CinAST核苷酸序列SEQ ID NO.5(人工合成)
ATGGAATCTGGTCTGGACCACAACTACAACAAAATCCTGGACATCCTGAAAGGTGCGATCAAAGGTGACGACAACCAGGTTAAAGCGCGTAAACACCTGCGTGTTGAACGTTGGCTGCGTGCGTACATCCAGCTGATCCGCCGCTTCGACGAAGAGGAACTGATCTTCTTCTCTGACATCTTCTCTGACAACTCTTGCTGGGACGGTATCAAACTGAAGAATAAAGCGGTTGGTGAACGTCTGACCGAAGAGAAGAACAAGAATGGTAAAGAAAACCCGCTGGACCTGGCGCGTCGTTACTACCTGGCGTGCAAATACTGCCTGGAAGACAAAATCCCGGGTCTGTTCGAACAGGTTTTCATGCGTTTCAAACGTTCTGCGTTCGAAGAAGACGGTTCTGACGACGACCTGCGTCGTCGTCTGCTGGAAAACATCGAAGAAACCTCTCCGATCCGTGCGTTCTGGTCTTTCCTGATCGACAAACAGATCGGTAAACTGAACGAATACAAATCTGTTGAAGGTCTGCAGAAATCTATCCAGATCAACTCTAACGATAATTGGGAAGAAGGTATCGAATTCTTCTACAACAAACTGCACAACGACTCTTCTATCTCTTCTCAGGACAAAGACGACCTGCTGATCGAAGCGGCGCTGTCTGCGGTTAAAGGTTACGAAGAAGTTGACACCATCGAATTCTGCCTGTCTAAAATGGACGACGAACAGAAGAAGAAACTGCTGGACCGTGACTACAAAGAAAACACCTACTACGCGGTTCTGAACGTTCTGGTTGGTCAGTACTACTTCGACTCTTTCATGGAACTGTCTCGTCTGTGCTCTCAGATCGAATGCGAAGAATACACCACCTTCCTGTCTTCTCTGTCTCGTCAGGTTCTGGAAAATCCGGACCTGTCTGAAGAAACCAAGAAGTGCATGATGAACGTTTGGGAACGTATCATCAAACTGAAAACCCAGGACCGTGGTGAACAGTCTATCTCTTCTATCTTCGTTGACTACTCTGTTACCTACACCATCGCGAACCTGATCGTTGACCCGTCTCGTCAGGGTGTTTCTAAAGAAGAAATCCTGGGTAAAATCCTGAAACACGTTAAAGAAATGTCTGGTGAAGAAATGATCAAAGTTAAAGACTCTGTTCTGTCTAAAATCCAGCTGTTCCACGGTGGTAAGAAGCTGCAGCTGGGTGAACAGGTTTTCTCTAAACTGGCGCAGGAAGCGTCTAAAGAATCTATCCTGCGTGAAGCGGGTGACACCCTGCCGCAGTCTTCTCTGTCTACCACCGACACCCCGTACAACATCAAATCTCTGTCTCACTCTAAA
突变蛋白CinAST氨基酸序列SEQ ID NO.6(人工合成)
MESGLDHNYNKILDILKGAIKGDDNQVKARKHLRVERWLRAYIQLIRRFDEEELIFFSDIFSDNSCWDGIKLKNKAVGERLTEEKNKNGKENPLDLARRYYLACKYCLEDKIPGLFEQVFMRFKRSAFEEDGSDDDLRRRLLENIEETSPIRAFWSFLIDKQIGKLNEYKSVEGLQKSIQINSNDNWEEGIEFFYNKLHNDSSISSQDKDDLLIEAALSAVKGYEEVDTIEFCLSKMDDEQKKKLLDRDYKENTYYAVLNVLVGQYYFDSFMELSRLCSQIECEEYTTFLSSLSRQVLENPDLSEETKKCMMNVWERIIKLKTQDRGEQSISSIFVDYSVTYTIANLIVDPSRQGVSKEEILGKILKHVKEMSGEEMIKVKDSVLSKIQLFHGGKKLQLGEQVFSKLAQEASKESILREAGDTLPQSSLSTTDTPYNIKSLSHSK
突变蛋白CinBST核苷酸序列SEQ ID NO.7(人工合成)
ATGCCGTCTAACGTTAAACCGCTGGAACTGGTTCAGCTGCTGCTGATGCGTAACAAATCTAAAGACGAATTCCTGGACTTCCAGAAACGTTTCCAGTCTTTCATCAACCAGTCTCCGTCTTTCCTGCACTCTGTTGGTAAACCGGGTTTCTTCCCGTCTTTCTTCTTCGGTATGTTCGCGACCGTTCTGGACACCGAACTGGCGACCAAAATCGGTATCAAGAAGCTGCACTTCCGTTTCGACGACAACCGTACCCTGAAAATCGCGATCCTGACCAACGAAGGTCTGAAATGCATCACCATGTCTGACCAGGTTGACGGTAACATGCACCTGAAATTCTCTCAGGGTGAACTGGAGAAGATCGCGCAGAAATGGAAAATGGGTGCGGAATTCGACAAACTGGAGAAGGAAGAACACGAAATCACCATCACCGGTAAAGAAGTTAAACACGGTAAAGTTGACCCGGCGTTCTCTAAGAAGACCGACTACTCTCAGAAAGGTTTCACCGAAATCGAGAAGGACCGTGACCAGCAGGACCTGGAATCTCTGATCTCTAAACTGTCTAACCAGCGTTTCGAAGAAGTTAAGAAGAACGCGGAACGTATGTTCAACTACATCACCAACGTTTACAAGAAGTACGAGAAGGAAACCCTGTTCTCTGGTAAAGAATCTTCTCACCACGGTTTCCTGGCGGGTTTCCTGATCAACTTCAAATACCGTTTCCACCTGAAACTGTACCTGGAACTGTTCGCGGGTAAAGGTTACGCGGACATCATCCTGCTGGTTCGTGGTTCTGACAAATCTCTGTCTTCTATCCCGATCATCATCGAACTGAAAGCGGGTACCGGTGAAATCTCTACCGTTATCAAAGCGCTGAAACAGGCGCAGGACTACGTTAAAGGTTCTTTCTCTAACTCTATCCGTATGATCACCATCGCGAACCGTGCGATCTGCGTTGGTCTGAACTTCGACATGGTTCACCACGAAAACGTTAAAATCCGTGTTGAAAACTTCCTGTCTCGTCGTGGTAACTCTGTTATCGAGGAACTGCTGGGTACCCGTGCGACCAACGCGGAAGTTATCCGTACCCAGCTGGAATACCTGTACTACGGTATCGTTTGGTCTAACGGTGGTTCTGACAACATCAACTACGTTTCTCGTATGATCCTGGGTCAGCTGGTTCTGATCTCTAACATCATCGATGAAGAGGAACTGGGTAAACACATCTTCATCTACGACCAGAACGACAAAATGGTTACCGGTTCTCAGAAACGTCCGGAAGCGGCGAAAGAATCTATCGAAGACTGCGTTACCACCATCGTTCTGACCCTGGGTAAGAAGGTTCTGATCCTGAACATCAACGAGAAGAACGAATTCGCGCTGCGTGTTCCGGACAACAAAGGTATCCCGATCGAAAACATCCGTCGTATCCAGAACGTTAACGACATCAAAATCCAGGAAATCACCTGCAACCTGTACTCTACCCCGTCTAACAAGAATCCGTTCGACCAGTACTGCAACAAGAATAAAGGTATCACCGTTAACACCTACGACTCTCTGGACAAATACAAACGTGGTAAAGAAATCCTGCAGGGTAACTTCACCCGTATCGTTGAAAACAAGAAGTTCAAAGCGGCGCTGTCTAAAGCGATCGAATCTGGTAAATACGACGACTACAAGAAGCTGTTCGAAGAAATCTCTCACATCCTGCACCCGTTCAAATCTCTGATCTCTAACGAAGCGACCTTCCAGGCGGTTCTGCACGGTCTGTTCTCTTCTTACGGTGAAGACAACATCAAAGTTATCACCGAATTCCAGATCGGTGGTGGTGAGAAGCTGGACGTTATGCTGGTTATCAACGCGACCGACCAGAAGAAGGAATACCCGCCGGTTGGTATCGAACTGAAATTCGCGAAGAAGGGTGAACTGGACAAGAAGGAGAAGGACGCGAAAGACCAGCTGAAACGTTACAAAGAAGGTGAAGCGTACAAAGTTATCACCGACGCGGGTAAAGTTAAACTGATCTACGCGGTTTTCAACAAAGGTGCGACCGACGAAGGTTCTCTGATCAAAATCGGTAACGAATTCGTTGAAGTTGACGTTCGTCACTCTTCTGTTGTTGCGTTCGGTCAGCAGCCGGGTTCTCTGCAGCAGCCGTACGTTAAACAGGCGGGTCTGTCTCGTGCGGTTAACCAG
突变蛋白CinBST氨基酸序列SEQ ID NO.8（人工合成）MPSNVKPLELVQLLLMRNKSKDEFLDFQKRFQSFINQSPSFLHSVGKPGFFPSFFFGMFATVLDTELATKIGIKKLHFRFDDNRTLKIAILTNEGLKCITMSDQVDGNMHLKFSQGELEKIAQKWKMGAEFDKLEKEEHEITITGKEVKHGKVDPAFSKKTDYSQKGFTEIEKDRDQQDLESLISKLSNQRFEEVKKNAERMFNYITNVYKKYEKETLFSGKESSHHGFLAGFLINFKYRFHLKLYLELFAGKGYADIILLVRGSDKSLSSIPIIIELKAGTGEISTVIKALKQAQDYVKGSFSNSIRMITIANRAICVGLNFDMVHHENVKIRVENFLSRRGNSVIEELLGTRATNAEVIRTQLEYLYYGIVWSNGGSDNINYVSRMILGQLVLISNIIDEEELGKHIFIYDQNDKMVTGSQKRPEAAKESIEDCVTTIVLTLGKKVLILNINEKNEFALRVPDNKGIPIENIRRIQNVNDIKIQEITCNLYSTPSNKNPFDQYCNKNKGITVNTYDSLDKYKRGKEILQGNFTRIVENKKFKAALSKAIESGKYDDYKKLFEEISHILHPFKSLISNEATFQAVLHGLFSSYGEDNIKVITEFQIGGGEKLDVMLVINATDQKKEYPPVGIELKFAKKGELDKKEKDAKDQLKRYKEGEAYKVITDAGKVKLIYAVFNKGATDEGSLIKIGNEFVEVDVRHSSVVAFGQQPGSLQQPYVKQAGLSRAVNQ
wMelCidA核苷酸序列SEQ ID NO.9(人工合成)
ATGCCGATCGAAACCAAACGTCAGGCGGAAGTTCTGAAGAAGCTGCAGGACGTTATCAAACACACCGACCGTGACATCGCGGCGGGTCGTAAACTGGCGATCAAACGTTGGGTTGAAACCTACATCGAATACATCAAACTGTTCAAAGACGACAAACTGGAATTCCTGTACAACGTTTTCCGTGACGAAGGTTGCTGGCTGGGTACCCGTCTGAACAACACCGTTCTGGGTCAGAAACTGACCGAAGAGAAGATCGGTGAAATCGACAACCCGCTGCCGCGTTACGGTATGGCGTCTCGTTACTGCATCACCGGTAAAATCGGTGACTTCTTCAACAAACAGTTCGTTCTGTCTCGTGGTCAGTTCACCTCTGAAGAAGTTGACTCTCAGGGTAACCCGATCTCTGACCAGTACGTTCGTAACATCCTGCTGTCTTCTATGAAACGTAACGGTCCGGTTTTCGACTTCTGGATCGACCGTGAATCTGGTGAACTGAAGAAGTACGACGCGGTTGAAGGTTTCGACTCTACCGTTAAACTGAAATGGTCTGAAGGTGTTGAATACTTCTACAACCAGCTGGAAGAGAAGGACAAAGAGAAGAAACTGACCGAAGCGATCGTTGCGCTGTCTCGTCCGCAGTCTGTTAAACGTGACGCGCCGATCCTGGACTTCTGCGTTCGTAACATCGGTGACAAAGACACCCTGCTGCAGAAACTGCTGCAGAAAGACAAAGGTGTTTACTTCCTGCTGGCGGAACTGATCGAATCTTGCTTCTTCGACACCGTTCACGACCTGGTTCAGTGCTGGTGCTACAAAGGTGTTTCTGCGGGTGGTGACTGCTCTGACAAAATCTTCTCTCAGCAGGACTACGAACTGTTCCTGTACTCTCTGTCTAACGTTATGCTGAAGAATCCGGAACTGTCTGTTCAGGCGCGTTCTCTGATCATGGAAATCTGGAAATGCGAACGTTTCGCGGAATACCGTGAAACCTCTGTTAACACCTCTAACTACACCGTTCCGATCAAATCTGTTCTGGGTGGTCTGATCATCAACTGGAAACGTGAAGACGTTTGCAAACCGGACCGTGAAATCGAGAAGGAAGAAATCCTGGACATGATCTCTTTCGCGAAAGGTTGCTTCCCGGAGAAGTTCGACCTGTTCAAAGAAGTTATGATCGAAAACCTGCGTATCTGCGGTCGTGAAGGTAAACGTAAAGGTGTTGACTACGGTAAATTCGCGGAAGAACTGTTCCTGCAGCTGGAGAAGGTTACCCTGCCGTCTGTTGGTGACGGTCCGTGGAACAACCTGCGTTCTCAGTCTAAAGTTTCTCTGCCGCTGGACGGTTCTGGTGACGGTCCGCAGTCTGAATTCGAAGCGCCGTCTGTTTCTGGTATCTCTGGTTCTCACAAGAAGCGTCGTATC
wPip CidA核苷酸序列SEQ ID NO.10(人工合成)
ATGCCGACCCAGAAAGAACTGCGTGACACCATGTCTAAGAAGCTGCAGGAAGCGATCAAACACCCGGACCCGGCGGTTGTTGCGGGTCGTAAATCTGCGATCAAACGTTGGGTTGGTGTTCTGCAGGACAACTTCATGGAACACATCAAATACTTCAAAGGTGACAAACTGAAATTCCTGCACAACGTTTTCCAGGACGAAGGTTGCTGGTCTGGTGTTCGTCTGGACAACGCGGCGCTGGGTCAGCGTTTCACCGAAGAGAAGATCGGTGGTATCGACAACCCGCTGCGTAAATACGAAATGGCGTGCTCTTACTGCGTTGTTGACAAAATCCACCCGCTGTTCCAGAAACGTTTCGAATCTTACCGTAACAAATTCCCGCCGGGTGCGTTCGACGGTAAAACCGAAACCGAATTCGGTAAATACGTTCGTAACTCTCTGCTGGACTCTATCAAACGTAAAGGTCCGGTTTTCGACTTCTGGATCGACCGTGAATCTGGTGAACTGAAGAAGTACGACGCGGTTGAAGGTTTCGACTCTGCGGTTAAATTCAAATGGTCTGAAGGTGTTGAATACTTCTACAACCACCTGAAAGAAGAAGACAAAGAGAAGAAACTGACCGAAGCGATCCTGGCGCTGTCTCGTGTTCAGTCTGTTGAGAAGGACGCGCCGATCCTGGACTTCTGCGTTAACAAAATCGTTGACAAAGACACCCTGCTGCAGAAACTGTCTCAGAAAGACAAAGGTGTTTACTCTCTGTTCGCGGAACTGATCGAATCTTGCTTCTTCGACACCGTTCACGACCTGGTTCAGTGCTGGTGCTACAAAGAAGTTTCTGCGGGTGGTGACCACTCTGAGAAGATCTTCTCTCAGCGTGACTACGAACTGTTCCTGTCTTCTCTGTCTGACACCATGCTGAAGAATCCGGAACTGTCTGTTCAGGCGCGTTCTCTGATCATGGAATTCTGGGAATGCGGTTCTCTGTACCAGTACCGTAAAGCGGCGGTTAACACCTCTAACTACACCGTTCCGACCTCTGGTGTTTTCGCGGAACTGATCGTTAACTGGCGTCGTGAAGACATCTACAAAACCGACGAAGAGAAGGAAATCGAGAAGAAAGAAATCCTGGACATGATGTCTTTCGCGAAAGACTGCTTCCCGGAGAAGTTCGAACTGTTCAAGAAGCTGATCATCCGTGACCTGCGTCTGTGCGGTCGTGAAGGTAAACGTGTTAACGTTGACTACGGTCTGTTCGCGGAAGAACTGTTCTCTGAACTGGAGAAGACCATCCTGCCGCCGGGTCCGGTTGGTGACGGTCCGTGCTCTAACCTGCGTTCTCGTTCTAAAGCGCACGGTTCTAAGAAGACCACCCTGCCGGTTGACGACTCTCCGCAGTCTGAACTGGGTACCCCGTCTGTTTCTGGTGTTTCTTCTTACAAGAAGAAATCTGTTTTCACCCTGTCTGGTAACAAA
wPip CidB核苷酸序列SEQ ID NO.11(人工合成)
ATGTCTAACGGTGACGGTCTGATCCGTTCTCTGGTTGACGGTGACCTGGAAGGTTTCCGTCAGGGTTTCGAATCTTTCCTGGACCAGTGCCCGTCTTTCCTGTACCACGTTTCTGCGGGTCGTTTCCTGCCGGTTTTCTTCTTCTCTATGTTCTCTACCGCGCACGACGCGAACATCCTGAACGCGAACGAACGTGTTTACTTCCGTTTCGACAACCACGGTGTTAACCCGCGTAACGGTGAAAACCGTAACACCGCGAACCTGAAAGTTGCGGTTTACCGTGACGGTCAGCAGGTTGTTCGTTGCTACTCTATCTCTGACCGTCCGAACTCTGACGGTCTGCGTTTCTCTACCCGTGAACGTAACGCGCTGGTTCAGGAAATCCGTCGTCAGAACCCGAACCTGCGTGAAGAAGACCTGAACTTCGAACAGTACAAAGTTTGCATGCACGGTAAAGGTAAATCTCAGGGTGAAGCGATCGCGACCGTTTTCGAAGTTATCCGTGAGAAGGACCGTCAGGGTCGTGACAAATTCGCGAAATACTCTGCGTCTGAAGTTCACTTCCTGCGTCAGCTGTTCCGTAACCACCGTCTGACCATCAAAGAAATCGAAGGTCGTCAGCTGAACCAGAACCAGCTGCGTCAGCTGGGTCGTTCTGTTAACTTCACCCGTGTTGAACCGGGTCAGCAGCGTATCGACAACTTCATGGAAATGCTGGCGTCTAACCAGCGTCAGGACGTTCGTGACTCTCTGCGTGGTGACATCCTGGAATACGTTACCGACACCTACAACAACTACCGTGCGCAGATCGAAAACAACATCGAAGGTCGTTCTCAGAAATTCGAATCTCACGGTTTCCTGCTGGGTTTCCTGGCGAACTTCTCTCACCGTTACACCATCGGTGTTGACCTGGACCTGTCTCCGCGTAACTCTCACGTTGCGTTCCTGGTTCGTCACCAGGTTGAACGTGAAAACATCCCGATCGTTATCAACCTGGCGACCCGTGCGCCGCCGTACATCGCGCTGAACCGTGCGCGTTCTCACGCGGAACGTCTGCACGTTTTCTCTTTCATCCCGATCCACACCGAATCTCGTAACACCGTTTGCGTTGGTCTGAACTTCAACCTGAACCTGGACCCGTTCTCTGTTGACACCGTTGGTCTGCAGCAGGACCGTTTCCCGCTGGTTCAGCGTCTGTTCGAATGCCTGGAAAACGAAGGTATCCGTGAAAACATCCGTGACTTCCTGCTGCACCACCTGCCGGCGGAAATCCCGCGTAACGCGGAAAACTACGACCGTATCTTCGACTGCATCACCGGTTTCGCGTTCGGTAACTCTGCGTTCGACCGTCACCCGCTGGAACTGGAAGAAGAAGACGAAGCGCCGATCACCAAATACATCTTCCGTCACGGTGACGAAGGTCTGCGTTGCCTGACCATGGTTTTCCACGCGGAAGGTTCTGACATCGTTATCCTGCACATCCGTGCGCACGACGCGCAGCAGCAGGGTGCGATCAACCTGCAGACCCTGAACGTTAACGGTAACGACGTTCACGTTTGGGAAGTTTCTTGCACCCTGAACAACCAGCTGGAACTGGACATCGACCTGCCGAACGACCTGGGTCTGTACCACGACTACCAGAACAACAACGCGAACAACTTCCTGGCGGGTGACCTGGTTCAGGTTCCGAACACCGAAAACGTTCACAACACCCTGAACCAGGTTGTTAACGACGGTTGGAAGAATATCGCGCAGCACCGTGGTCTGTTCCAGGAAATCTCTGGTGCGCTGATGCCGCTGGTTGACACCATCAACGTTAACTCTGAAGACAAATTCCGTTCTATCCTGCACGGTACCTTCTACGCGTCTGACAACCCGTACAAAGTTCTGGCGATGTACAAAGTTGGTCAGACCTACTCTCTGAAACGTGGTCAGGAAGAAGAAGGTGAACGTGTTATCCTGACCCGTATCACCGAACAGCGTCTGGACCTGCTGCTGCTGCGTCAGCCGCGTGAAAACGACCTGGACACCCACCCGATCGGTTACGTTCTGCGTCTGGCGAACAACGCGGAAGAAGTTGGTCAGCAGCAGAACGACGCGCGTCAGGAAATCGGTCGTCTGAAGAAGCAGCACCGTGGTTTCATCCCGATCACCTCTGGTAACGAAGTTGTTCTGTTCCCGATCGTTTTCAACCGTGACGCGCACGAAGCGGGTAACCTGATCCTGTTCCCGGAAGGTATCGGTCGTGAAGAACACGTTCACCGTCTGGACCGTCACGTTCGTTCTTCTCGTCCGGGTGGTCTGGTTGGTCCGGAATCTGTTATCGACGAAAACCCGCCGGAAGGTCTGCTGTCTGACCAGACCCGTGAAAACTTCCGTCGTTTCTACGAAGAGAAGGCGCCGGGTCAGAACTCTATCTTCCTGCTGGACATCGGTGACAACCTGCACGTTCCGTTCTCTTACCTGCAGGGTACCCGTGCGCAGGTTATCGAAACCCTGAAATCTCGTATCCGTGGTGGTGGTACCCCGACCGCGCAGGGTATCCTGCAGCAGATCAACGCGATCCTGCGTCGTAACAACGCGCGTGAAATCGAAGACGTTCACGACCTGCTGGCGCTGGACTTCGCGACCGACAACCAGAACTACCGTTACTGGCTGCAGACCCACGACATGTTCTTCGCGGCGCGTCAGTACACCTTCCTGGACAACCAGTCTCACTCTACCAACGACCACTACGGTTTCGAAATCACCTCTGTTGGTGTTGACGGTAACCAGAACGACCCGACCGGTCGTGGTCTGCTGTCTTCTCACATCACCAACTTCAAACAGAAAGTTGACTCTGGTGAGAAGGACCGTCTGATCGCGATCATCAACGTTGGTAACCGTCACTGGGTTACCCTGGTTATCGTTCACCAGAACGGTAACTACTACGGTTACTACGCGGACTCTCTGGGTCCGGACTCTGGTATCGACAACAACATCCGTGGTGCGCTGCGTGAATGCGACATCAACGACGACAACGTTCACAACATCTCTGTTCACCAGCAGACCGACGGTCACAACTGCGGTATCTGGGTTTACGAAAACGCGCGTGACATCAACCAGGCGATCGACCAGGCGCTGCAGGGTAACAACAACTTCGGTGAGAAGGGTGAAGGTATCATCGGTTACATCCGTGGTCTGCTGTCTGCGGGTATCGGTAACGACACCCGTCAGCCGCGTCGTAACGAACAGTACTTCGAAGACCGTCGTCGTGACATCTCTCAGCTGCTGCAGAACGACCCGAACCTGCCGTCTCGTCGTTCTGACCTGATCCAGGCGCACCCGGGTATCCAGCACGAAATCGACCCGCTGCTGCTGCAGTTCCTGGGTCTGCAGTACCCGCAGCGTGGTGGTGGTGGTGCGCTGCAGCTGGGTGGTGAACGTGTTATCTCTATCGACTTCGGTAACCCGCAGTCTGCGCTGGACAAAATCGACGGTGTTTCTCGTGTTTACAACCACTCTAACTCTCGTGGTTCTCGT
wPip CidB氨基酸序列SEQ ID NO.12(人工合成)
MSNGDGLIRSLVDGDLEGFRQGFESFLDQCPSFLYHVSAGRFLPVFFFSMFSTAHDANILNANERVYFRFDNHGVNPRNGENRNTANLKVAVYRDGQQVVRCYSISDRPNSDGLRFSTRERNALVQEIRRQNPNLREEDLNFEQYKVCMHGKGKSQGEAIATVFEVIREKDRQGRDKFAKYSASEVHFLRQLFRNHRLTIKEIEGRQLNQNQLRQLGRSVNFTRVEPGQQRIDNFMEMLASNQRQDVRDSLRGDILEYVTDTYNNYRAQIENNIEGRSQKFESHGFLLGFLANFSHRYTIGVDLDLSPRNSHVAFLVRHQVERENIPIVINLATRAPPYIALNRARSHAERLHVFSFIPIHTESRNTVCVGLNFNLNLDPFSVDTVGLQQDRFPLVQRLFECLENEGIRENIRDFLLHHLPAEIPRNAENYDRIFDCITGFAFGNSAFDRHPLELEEEDEAPITKYIFRHGDEGLRCLTMVFHAEGSDIVILHIRAHDAQQQGAINLQTLNVNGNDVHVWEVSCTLNNQLELDIDLPNDLGLYHDYQNNNANNFLAGDLVQVPNTENVHNTLNQVVNDGWKNIAQHRGLFQEISGALMPLVDTINVNSEDKFRSILHGTFYASDNPYKVLAMYKVGQTYSLKRGQEEEGERVILTRITEQRLDLLLLRQPRENDLDTHPIGYVLRLANNAEEVGQQQNDARQEIGRLKKQHRGFIPITSGNEVVLFPIVFNRDAHEAGNLILFPEGIGREEHVHRLDRHVRSSRPGGLVGPESVIDENPPEGLLSDQTRENFRRFYEEKAPGQNSIFLLDIGDNLHVPFSYLQGTRAQVIETLKSRIRGGGTPTAQGILQQINAILRRNNAREIEDVHDLLALDFATDNQNYRYWLQTHDMFFAARQYTFLDNQSHSTNDHYGFEITSVGVDGNQNDPTGRGLLSSHITNFKQKVDSGEKDRLIAIINVGNRHWVTLVIVHQNGNYYGYYADSLGPDSGIDNNIRGALRECDINDDNVHNISVHQQTDGHNCGIWVYENARDINQAIDQALQGNNNFGEKGEGIIGYIRGLLSAGIGNDTRQPRRNEQYFEDRRRDISQLLQNDPNLPSRRSDLIQAHPGIQHEIDPLLLQFLGLQYPQRGGGGALQLGGERVISIDFGNPQSALDKIDGVSRVYNHSNSRGSR
正向引物F1核苷酸序列SEQ ID NO.13(人工合成)
GGAATTCCATATGCCAATCGAAACCAAGAAACAA
反向引物R1核苷酸序列SEQ ID NO.14(人工合成)
CCGCTCGAGGATGCGGCGTTTTTTATGGCTG
正向引物F2核苷酸序列SEQ ID NO.15(人工合成)
CGGGATCCATGGACTATAAAGACGATGACGATAAAATGCCAATCGAAACCAAGAAAC
反向引物R2核苷酸序列SEQ ID NO.16(人工合成)
ACGCGTCGACTTAGATGCGGCGTTTTTTATGGCTG
wMel CidB核苷酸序列SEQ ID NO.17(人工合成)
ATGGACGGTGACCTGGACGGTTTCCGTCAGGAATTCGAATCTTTCCTGGACCAGTGCCCGTTCTTCCTGTACCACGTTTCTACCGGTCGTTTCCTGCCGGTTTTCTTCTTCTCTATGTTCGCGACCGCGCACGACGCGAACATCCTGAAAGCGAACGAACGTGTTTACTTCCGTTTCGACAACCACGGTATCGACACCGGTGGTCGTAACCGTAACACCGGTAACCTGAAAGTTGCGGTTTACCACGACGGTCAGCAGGTTGTTCGTTGCTACTCTATCTCTGACCGTCTGAACTCTGACGGTCTGCGTTTCTCTACCCGTGAACGTAACGCGCTGGTTCGTGAAATCCGTGGTCAGAACCCGAACCTGCGTGAAGAAGACCTGAACTTCGAACAGTACAAAGTTTGCATGCACGGTAAAGGTAAATCTCAGGGTGAAGCGATCGCGACCGTTTTCGAAGTTATCCGTGAGAAGGACTCTCAGGGTCGTGACCGTTTCGCGAAATACTCTGCGTCTGAAATCTCTCTGCTGCGTCACATCGAACGTAACCGTCTGAACGGTATCAACGCGCCGGCGCCGCGTTCTCTGCTGACCGTTAAAGAAATCGGTTCTATCCGTCTGAACCAGGACCAGCGTGTTCAGCTGGGTCACCTGGTTAACTTCGTTCAGGTTGCGCCGGGTCAGCAGGGTATCTTCTCTTTCATGGAAGTTCTGGCGTCTAACCAGAAAATCAACATCGAACGTGGTATCAACGAAGGTATCCTGCCGTACATCACCCGTATCTACCGTTCTTACCTGGGTTCTCTGCAGAACGACATCCAGAACCGTTCTCAGAAATTCGAATCTCACGGTTTCTTCCTGGGTCTGCTGGCGAACTTCATCCACCTGTACACCATCGACATCGACCTGGACCTGTCTCCGGGTAACTCTTACGTTGCGTTCCTGATCTGCCACCAGGCGGAACGTGAAAACATCCCGATCGTTATCAACGTTACCCGTTGGCGTACCTCTTCTGACATCGCGCTGAACCGTGCGCGTGCGGACGCGAAACGTCTGCACGTTTCTTCTTTCATCTCTATCCACACCGAATCTCGTAACGCGGTTTGCATCGGTCTGAACTTCAACCTGAACATCGACCCGTTCTCTATCGACACCGTTGAATTCCTGGAAAACCGTTTCCCGCTGGTTCAGCGTCTGTTCGAATGCCTGGAAGACGAAGGTATCCGTGAAAACATCCGTGACTTCCTGCTGCAGCACCTGCCGAACGAAATCCCGCGTAACGCGGAAAACTACAACCGTATCTTCGACTGCATCACCGGTTTCGCGTTCGGTAACTCTATCCTGGAAGAATTCCGTCTGGTTAACGCGGTTCAGCAGCGTGTTCGTAAATACATCTTCCGTTACGGTGACGAAAACCACGCGCTGACCATGGTTTTCCACACCCAGGGTTCTGACATCGTTATCCTGCACATCCGTGACAACAACGCGGTTCAGCAGGGTGCGATCAACCTGCAGGACCTGAACGTTGACGGTAACAACGTTCACGTTCGTGAAGTTTCTTGCACCCTGAACAACCAGCTGGGTCTGAACATCCACACCGACAACCTGGGTCTGTACCACAACTACCAGAACAACAACGCGAACAACTTCCTGGGTGGTAACCTGGTTCAGGTTCCGAACGCGGGTAACGTTCACAACGCGCTGAACCAGGTTATGAACGACGGTTGGCAGGACCGTTTCCAGCACCAGGAACTGTTCCGTAACATCTCTGCGGTTCTGATGCCGGAAGACACCCACGGTAACATGATCATCGACGTTAACTCTAAAGACAAATTCCGTTCTATCCTGCACGGTACCTTCTACGCGTCTGACAACCCGTACAAAGTTCTGGCGATGTACAAAGTTGGTCAGACCTACTCTCTGAAACGTTGGCAGGAAGAAGAAGGTGAACGTGTTATCCTGACCCGTGTTACCGAACAGCGTCTGGGTCTGCTGCTGCTGCGTCAGCCGACCGCGGACACCCACCCGATCGGTTACGTTCTGGGTTTCGCGGACAACGCGGAAGAAGTTGAACAGGAACAGGACGAAGCGCGTTACAAAATCACCGAACTGATGTCTAAACAGCGTGGTTACCTGCCGATCACCTCTGGTAACGAAGTTGTTCTGTCTTACGCGGTTTTCAACCGTGGTGCGCAGCGTGCGGAAGACTTCATCTCTCTGCCGCAGCAGGCGGTTTACGTTCACCGTCTGGACCGTCGTGGTCACGACTCTCGTCCGGAAGTTCTGGTTGGTCCGGAATCTGTTATCGACGAAAACCCGCCGGAAAACCTGCTGTCTGACCAGACCCGTGAAAACTTCCGTCGTTTCTACATGGAGAAGCGTCCGGGTCAGAACTCTATCTTCCTGCTGGACATCGACGACAACCTGCACGTTCCGTTCTCTTACCTGCAGGGTACCCGTGCGCAGGCGATCGAAACCCTGCGTTCTCGTATCCGTGGTGGTGGTACCTCTACCGCGCAGGGTATCCTGCAGCAGATCAACACCATCCTGCGTCGTAACAACGCGCGTGAAATCGAAGACGTTCACAACCTGCTGGCGCTGGACTTCGCGACCGAAAACCAGAACTTCCGTTACTGGCTGCAGACCCACGACATGTTCTTCGCGGCGCGTCAGTACACCTTCCACGACGACCGTTCTAACCCGACCAACGACCGTCACGACTTCGCGATCACCTCTGTTGGTGTTGACGGTAACCAGAACGACCCGACCGGTCGTGACCTGCTGTCTTCTAACATCGACAACTTCAAACAGAAAGTTGACTCTGGTGAGAAGGACCGTCTGACCGCGATCATCAACGTTGGTAACCGTCACTGGGTTACCCTGGTTATCGTTCACCAGAACGGTAACTACTACGGTTACTACGCGGACTCTCTGGGTCCGGACTCTCGTATCGACAACAACATCCGTGGTGCGCTGCGTGAATGCGACATCTCTGACGACAACGTTCACGACGTTTCTGTTCACCAGCAGACCGACGGTCACAACTGCGGTATCTGGGCGTACGAAAACGCGCGTGACATCAACCAGGCGATCGACCAGGCGCTGCAGGGTAACTCTAACTTCGGTGAGAAGGGTGAAGGTATCATCGGTTACATCCGTGGTCTGCTGTCTGCGGGTATCGGTAACGACACCCGTCAGCCGCAGCGTAACGAACAGTACTTCCGTAACCGTCGTCGTAACATCTCTCAGCTGTTCCAGAACGACTCTCTGTCTTCTCCGCGTGGTCGTCTGATCCAGGGTCGTCCGGGTATCCAGCACGAAATCGACCCGCTGCTGCTGCAGTTCCTGGAACTGCAGTACCCGCAGCGTGGTGGTGGTGGTGCGCTGCAGCTGGGTGGTGAACGTGTTATCTCTATCGACTTCGGTCCGCAGTCTGTTCTGGACGAAATCGACGGTGTTAACCGTGTTTACGACCACTCTAACGGTCGTGGTTCTCGT
正向引物F3核苷酸序列SEQ ID NO.18(人工合成)
GGAATTCCATATGTCTAACGGTGACGGTCTGA
反向引物R3核苷酸序列SEQ ID NO.19(人工合成)
CCGCTCGAGACGAGAACCACGAGAGTTAGAG
正向引物F4核苷酸序列SEQ ID NO.20(人工合成)
CGGGATCCATGGACTATAAAGACGATGACGATAAAATGTCTAACGGTGACGGTCTGA
反向引物R4核苷酸序列SEQ ID NO.21(人工合成)
ACGCGTCGACTTAACGAGAACCACGAGAGTTAGAG
wPip CinA核苷酸序列SEQ ID NO.22(人工合成)
ATGGAATCTGGTCTGGACCACAACTACAACAAAATCCTGGACATCCTGAAAGGTGCGATCAAAGGTGACGACAACCAGGTTAAAGCGCGTAAACACCTGCGTGTTGAACGTTGGCTGCGTGCGTACATCCAGCTGATCGAAGACTTCGACGAAGAGAAGCTGATCTTCTTCTCTGACATCTTCTCTGACAACTCTTGCTGGGACGGTATCAAACTGAAGAATAAAGCGGTTGGTGAACGTCTGACCGAAGAGAAGAACAAGAATGGTAAAGAAAACCCGCTGGACCTGGCGGACCGTTACTACCTGGCGTGCAAATACTGCCTGGAAGACAAAATCCCGGGTCTGTTCGAACAGGTTTTCATGCGTTTCAAACGTTCTGCGTTCGAAGAAGACGGTTCTGACGACGACCTGCGTCGTGAACTGCTGGAAAACATCGAAGAAACCTCTCCGATCGAAGCGTTCTGGTCTTTCCTGATCGACAAACAGATCGGTAAACTGAACGAATACAAATCTGTTGAAGGTCTGCAGAAATCTATCCAGATCAACTCTAACAAGAATTGGGAAGAAGGTATCGAATTCTTCTACAACAAACTGCACAACGACTCTTCTATCTCTTCTCAGGACAAAGACGACCTGCTGATCGAAGCGGCGCTGTCTGCGGTTAAAGGTTACAAAGAAGTTGACACCATCGAATTCTGCCTGTCTAAAATGGACGACGAACAGAAGAAGAAACTGCTGGACCGTGACTACAAAGAAAACACCTACTACGCGGTTCTGAACGTTCTGGTTGGTCAGTACTACTTCGACTCTTTCATGGAACTGTCTCGTCTGTGCTCTCAGATCGAATGCGAACGTTACACCACCTTCCTGTCTTCTCTGTCTGACCAGGTTCTGAAGAATCCGGACCTGTCTGAAGAAACCAAGAAGTGCATGATGAACGTTTGGGAACGTATCATCAAACTGAAAACCCAGGACCGTGGTGAACAGTCTATCTCTTCTATCTTCGTTGACTACTCTGTTACCTACACCATCGCGAACCTGATCGTTGACCCGTCTCGTCAGGGTGTTTCTAAAGAAGAAATCCTGGGTAAAATCCTGAAACACGTTAAAGAAATGTCTGGTGAAGAAATGATCAAAGTTAAAGACTCTGTTCTGTCTAAAATCCAGCTGTTCCACGGTGGTAAGAAGCTGCAGCTGGGTGAACAGGTTTTCTCTAAACTGGCGCAGGAAGCGTCTAAAGAATCTATCCTGCGTGAAGCGGGTGACACCCTGCCGCAGTCTTCTCTGTCTACCACCGACACCCCGTACAACATCAAATCTCTGTCTCACTCTAAA
正向引物F5核苷酸序列SEQ ID NO.23(人工合成)
GGAATTCCATATGGAATCTGGTCTGGAC
反向引物R5核苷酸序列SEQ ID NO.24(人工合成)
CCGCTCGAGTTTAGAGTGAGACAGAGATTTG
正向引物F6核苷酸序列SEQ ID NO.25(人工合成)
CGGGATCCATGGACTATAAAGACGATGACGATAAAATGGAATCTGGTCTGGAC
反向引物R6核苷酸序列SEQ ID NO.26(人工合成)
ACGCGTCGACTTATTTAGAGTGAGACAGAGATTTG
wPip CinB核苷酸序列SEQ ID NO.27(人工合成)
ATGCCGTCTAACGTTAAACCGCTGGAACTGGTTCAGCTGCTGCTGATGCGTAACAAATCTAAAGACGAATTCCTGGACTTCCAGAAACGTTTCCAGTCTTTCATCAACCAGTCTCCGTCTTTCCTGCACTCTGTTGGTAAACCGGGTTTCTTCCCGTCTTTCTTCTTCGGTATGTTCGCGACCGTTCTGGACACCGAACTGGCGACCAAAATCGGTATCAAGAAGCTGCACTTCCGTTTCGACGACAACCGTACCCTGAAAATCGCGATCCTGACCAACGAAGGTCTGAAATGCATCACCATGTCTGACCAGGTTGACGGTAACATGCACCTGAAATTCTCTCAGGGTGAACTGGAGAAGATCGCGCAGAAATGGAAAATGGGTGCGGAATTCGACAAACTGGAGAAGGAAGAACACGAAATCACCATCACCGGTAAAGAAGTTAAACACGGTAAAGTTGACCCGGCGTTCTCTAAGAAGACCGACTACTCTCAGAAAGGTTTCACCGAAATCGAGAAGGACCGTGACCAGCAGGACCTGGAATCTCTGATCTCTAAACTGTCTAACCAGGACTTCGAAGAAGTTAAGAAGAACGCGCGTCGTATGTTCAACTACATCACCAACGTTTACAAGAAGTACGAGAAGGAAACCCTGTTCTCTGGTAAAGAATCTTCTCACCACGGTTTCCTGGCGGGTTTCCTGATCAACTTCAAATACCGTTTCCACCTGAAACTGTACCTGGAACTGTTCGCGGGTAAAGGTTACGCGGACATCATCCTGCTGGTTCGTGGTTCTGACAAATCTCTGTCTTCTATCCCGATCATCATCGAACTGAAAGCGGGTACCGGTGAAATCTCTACCGTTATCAAAGCGCTGAAACAGGCGCAGGACTACGTTAAAGGTTCTTTCTCTAACTCTATCCGTATGATCACCATCGCGAACGAAGCGATCTGCGTTGGTCTGAACTTCGACATGGTTCACCACGAAAACGTTAAAATCGACGTTGAAAACTTCCTGTCTCGTGAAGGTAACTCTGTTATCGAGAAGCTGCTGGGTACCGAAGCGACCAACGCGGAAGTTATCCGTACCCAGCTGGAATACCTGTACTACGGTATCGTTTGGTCTAACGGTGGTTCTGACAACATCAACTACGTTTCTCGTATGATCCTGGGTCAGCTGGTTCTGATCTCTAACATCATCAAACGTGAGAAGCTGGGTAAACACATCTTCATCTACGACCAGAACGACAAAATGGTTACCGGTTCTCAGAAACGTCCGGAAGCGGCGAAAGAATCTATCGAAGACTGCGTTACCACCATCGTTCTGACCCTGGGTAAGAAGGTTCTGATCCTGAACATCAACGAGAAGAACGAATTCGCGCTGCGTGTTCCGGACAACAAAGGTATCCCGATCGAAAACATCCGTCGTATCCAGAACGTTAACGACATCAAAATCCAGGAAATCACCTGCAACCTGTACTCTACCCCGTCTAACAAGAATCCGTTCGACCAGTACTGCAACAAGAATAAAGGTATCACCGTTAACACCTACGACTCTCTGGACAAATACAAACGTGGTAAAGAAATCCTGCAGGGTAACTTCACCCGTATCGTTGAAAACAAGAAGTTCAAAGCGGCGCTGTCTAAAGCGATCGAATCTGGTAAATACGACGACTACAAGAAGCTGTTCGAAGAAATCTCTCACATCCTGCACCCGTTCAAATCTCTGATCTCTAACGAAGCGACCTTCCAGGCGGTTCTGCACGGTCTGTTCTCTTCTTACGGTGAAGACAACATCAAAGTTATCACCGAATTCCAGATCGGTGGTGGTGAGAAGCTGGACGTTATGCTGGTTATCAACGCGACCGACCAGAAGAAGGAATACCCGCCGGTTGGTATCGAACTGAAATTCGCGAAGAAGGGTGAACTGGACAAGAAGGAGAAGGACGCGAAAGACCAGCTGAAACGTTACAAAGAAGGTGAAGCGTACAAAGTTATCACCGACGCGGGTAAAGTTAAACTGATCTACGCGGTTTTCAACAAAGGTGCGACCGACGAAGGTTCTCTGATCAAAATCGGTAACGAATTCGTTGAAGTTGACGTTCGTCACTCTTCTGTTGTTGCGTTCGGTCAGCAGCCGGGTTCTCTGCAGCAGCCGTACGTTAAACAGGCGGGTCTGTCTCGTGCGGTTAACCAG
正向引物F7核苷酸序列SEQ ID NO.28(人工合成)
GGAATTCCATATGCCGTCTAACGTTAAACCG
反向引物R7核苷酸序列SEQ ID NO.29(人工合成)
CCGCTCGAGCTGGTTAACCGCACGAGACA
正向引物F8核苷酸序列SEQ ID NO.30(人工合成)
CGGGATCCATGGACTATAAAGACGATGACGATAAAATGCCGTCTAACGTTAAACCG
反向引物R8核苷酸序列SEQ ID NO.31(人工合成)
ACGCGTCGACTTACTGGTTAACCGCACGAGACA
正向引物F9核苷酸序列SEQ ID NO.32(人工合成)
CGGGATCCATGGACTATAAAGACGATGACGATAAAATGTCTAACGGTGACGGTCTGA
反向引物R9核苷酸序列SEQ ID NO.33(人工合成)
ACGCGTCGACTTAACGAGAACCACGAGAGTTAGAG
wPip CinAST(I-III)氨基酸序列SEQ ID NO.34(人工合成)
MESGLDHNYNKILDILKGAIKGDDNQVKARKHLRVERWLRAYIQLIEDFDEEKLIFFSDIFSDNSCWDGIKLKNKAVGERLTEEKNKNGKENPLDLARRYYLACKYCLEDKIPGLFEQVFMRFKRSAFEEDGSDDDLRRRLLENIEETSPIRAFWSFLIDKQIGKLNEYKSVEGLQKSIQINSNDNWEEGIEFFYNKLHNDSSISSQDKDDLLIEAALSAVKGYEEVDTIEFCLSKMDDEQKKKLLDRDYKENTYYAVLNVLVGQYYFDSFMELSRLCSQIECERYTTFLSSLSDQVLKNPDLSEETKKCMMNVWERIIKLKTQDRGEQSISSIFVDYSVTYTIANLIVDPSRQGVSKEEILGKILKHVKEMSGEEMIKVKDSVLSKIQLFHGGKKLQLGEQVFSKLAQEASKESILREAGDTLPQSSLSTTDTPYNIKSLSHSK
wPip CinAST(I-III)核苷酸序列SEQ ID NO.35(人工合成)
ATGGAATCTGGTCTGGACCACAACTACAACAAAATCCTGGACATCCTGAAAGGTGCGATCAAAGGTGACGACAACCAGGTTAAAGCGCGTAAACACCTGCGTGTTGAACGTTGGCTGCGTGCGTACATCCAGCTGATCGAAGACTTCGACGAAGAGAAGCTGATCTTCTTCTCTGACATCTTCTCTGACAACTCTTGCTGGGACGGTATCAAACTGAAGAATAAAGCGGTTGGTGAACGTCTGACCGAAGAGAAGAACAAGAATGGTAAAGAAAACCCGCTGGACCTGGCGCGTCGTTACTACCTGGCGTGCAAATACTGCCTGGAAGACAAAATCCCGGGTCTGTTCGAACAGGTTTTCATGCGTTTCAAACGTTCTGCGTTCGAAGAAGACGGTTCTGACGACGACCTGCGTCGTCGTCTGCTGGAAAACATCGAAGAAACCTCTCCGATCCGTGCGTTCTGGTCTTTCCTGATCGACAAACAGATCGGTAAACTGAACGAATACAAATCTGTTGAAGGTCTGCAGAAATCTATCCAGATCAACTCTAACGATAATTGGGAAGAAGGTATCGAATTCTTCTACAACAAACTGCACAACGACTCTTCTATCTCTTCTCAGGACAAAGACGACCTGCTGATCGAAGCGGCGCTGTCTGCGGTTAAAGGTTACGAAGAAGTTGACACCATCGAATTCTGCCTGTCTAAAATGGACGACGAACAGAAGAAGAAACTGCTGGACCGTGACTACAAAGAAAACACCTACTACGCGGTTCTGAACGTTCTGGTTGGTCAGTACTACTTCGACTCTTTCATGGAACTGTCTCGTCTGTGCTCTCAGATCGAATGCGAACGTTACACCACCTTCCTGTCTTCTCTGTCTGACCAGGTTCTGAAGAATCCGGACCTGTCTGAAGAAACCAAGAAGTGCATGATGAACGTTTGGGAACGTATCATCAAACTGAAAACCCAGGACCGTGGTGAACAGTCTATCTCTTCTATCTTCGTTGACTACTCTGTTACCTACACCATCGCGAACCTGATCGTTGACCCGTCTCGTCAGGGTGTTTCTAAAGAAGAAATCCTGGGTAAAATCCTGAAACACGTTAAAGAAATGTCTGGTGAAGAAATGATCAAAGTTAAAGACTCTGTTCTGTCTAAAATCCAGCTGTTCCACGGTGGTAAGAAGCTGCAGCTGGGTGAACAGGTTTTCTCTAAACTGGCGCAGGAAGCGTCTAAAGAATCTATCCTGCGTGAAGCGGGTGACACCCTGCCGCAGTCTTCTCTGTCTACCACCGACACCCCGTACAACATCAAATCTCTGTCTCACTCTAAA
序列表
<110> 天津大学
<120> CifA和CifB突变蛋白用于调控胞质不相容性的用途
<130> HP210682LZ
<160> 35
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1422
<212> DNA
<213> 人工序列()
<400> 1
atgccaatcg aaaccaagaa acaagccgag gtgctgaaga agctgcaaga tgtgatcaag 60
cataccgatc gcgatatcgc ggccggccgt aaactggcca ttaaacgctg ggtggagacg 120
tatattgaat atatcaaata ctttaaggac gacaagctgg agtttctcta caacgtgttc 180
cgtgacgagg gctgttggct gggcacccgc ctcaacaaca ccgtgctcgg tcagaaactc 240
acggaggaga aaatcggcga gatcgataac ccgctgcgcc gttacggcat ggccagtcgt 300
tactgcatca ccggcaaaat ccacccgctg ttccaaaaac gcttcgagag ttaccgcaat 360
aagttcccac cgggtgcctt cgacggcaag accgagacgg aattcggcaa gtacgttcgt 420
aacagtctgc tggatagcat caagcgcaaa ggcccggttt tcgacttttg gatcgaccgc 480
gagagcggcg agctgaagaa atacgatgcc gtggagggct ttgacagcac ggtgaagctg 540
aagtggagcg agggtgttga gtacttctac aaccagctgg aagagaagga taaggagaag 600
aaactgaccg aagccatcgt ggcgctgagt cgcccacaga gtgttaaacg cgatgcgccg 660
attctggatt tctgcgtgcg caatatcggc gacaaagata cgctgctgca gaaactgctg 720
cagaaggaca aaggcgtgta ttttctgctc gccgagctca tcgagagctg tttctttgac 780
accgttcacg atctggttca gtgctggtgc tacaaaggtg ttagcgcggg tggcgattgc 840
agcgacaaaa tcttcagcca gcgcgactac gagctgtttc tgagcagtct gagcgatgtg 900
atgctgaaga atccggaact cagcgtgcaa gcccgtagtc tgatcatgga gatttggaag 960
tgcgaacgct tcgcggaata ccgcgagacg agtgtgaaca cgagcaacta caccgtgccg 1020
atcaaaagcg tgctgggcga actgatcatc aactggaaac gcgaagacgt gtgtaaaccg 1080
gatcgcgaga tcgaaaaaga agaaattctg gatatgatca gcttcgcgaa gggctgcttc 1140
ccggagaaat tcgatctgtt caaagaagtt atgatccgca atctccgtct gtgcggtcgc 1200
gagggcaaac gcaagggtgt ggattacggt aagttcgcgg aggaactctt cctccagctg 1260
gaaaaggtta cgctgccgag cgtgggcgat ggtccgtgga ataatctgcg cagccagagc 1320
aaagtgagtc tgccgctgga tggcagtggc gacggtccac aaagtgaatt cgaagccccg 1380
agtgtgagcg gcattagcgg cagccataaa aaacgccgca tc 1422
<210> 2
<211> 474
<212> PRT
<213> 人工序列()
<400> 2
Met Pro Ile Glu Thr Lys Lys Gln Ala Glu Val Leu Lys Lys Leu Gln
1 5 10 15
Asp Val Ile Lys His Thr Asp Arg Asp Ile Ala Ala Gly Arg Lys Leu
20 25 30
Ala Ile Lys Arg Trp Val Glu Thr Tyr Ile Glu Tyr Ile Lys Tyr Phe
35 40 45
Lys Asp Asp Lys Leu Glu Phe Leu Tyr Asn Val Phe Arg Asp Glu Gly
50 55 60
Cys Trp Leu Gly Thr Arg Leu Asn Asn Thr Val Leu Gly Gln Lys Leu
65 70 75 80
Thr Glu Glu Lys Ile Gly Glu Ile Asp Asn Pro Leu Arg Arg Tyr Gly
85 90 95
Met Ala Ser Arg Tyr Cys Ile Thr Gly Lys Ile His Pro Leu Phe Gln
100 105 110
Lys Arg Phe Glu Ser Tyr Arg Asn Lys Phe Pro Pro Gly Ala Phe Asp
115 120 125
Gly Lys Thr Glu Thr Glu Phe Gly Lys Tyr Val Arg Asn Ser Leu Leu
130 135 140
Asp Ser Ile Lys Arg Lys Gly Pro Val Phe Asp Phe Trp Ile Asp Arg
145 150 155 160
Glu Ser Gly Glu Leu Lys Lys Tyr Asp Ala Val Glu Gly Phe Asp Ser
165 170 175
Thr Val Lys Leu Lys Trp Ser Glu Gly Val Glu Tyr Phe Tyr Asn Gln
180 185 190
Leu Glu Glu Lys Asp Lys Glu Lys Lys Leu Thr Glu Ala Ile Val Ala
195 200 205
Leu Ser Arg Pro Gln Ser Val Lys Arg Asp Ala Pro Ile Leu Asp Phe
210 215 220
Cys Val Arg Asn Ile Gly Asp Lys Asp Thr Leu Leu Gln Lys Leu Leu
225 230 235 240
Gln Lys Asp Lys Gly Val Tyr Phe Leu Leu Ala Glu Leu Ile Glu Ser
245 250 255
Cys Phe Phe Asp Thr Val His Asp Leu Val Gln Cys Trp Cys Tyr Lys
260 265 270
Gly Val Ser Ala Gly Gly Asp Cys Ser Asp Lys Ile Phe Ser Gln Arg
275 280 285
Asp Tyr Glu Leu Phe Leu Ser Ser Leu Ser Asp Val Met Leu Lys Asn
290 295 300
Pro Glu Leu Ser Val Gln Ala Arg Ser Leu Ile Met Glu Ile Trp Lys
305 310 315 320
Cys Glu Arg Phe Ala Glu Tyr Arg Glu Thr Ser Val Asn Thr Ser Asn
325 330 335
Tyr Thr Val Pro Ile Lys Ser Val Leu Gly Glu Leu Ile Ile Asn Trp
340 345 350
Lys Arg Glu Asp Val Cys Lys Pro Asp Arg Glu Ile Glu Lys Glu Glu
355 360 365
Ile Leu Asp Met Ile Ser Phe Ala Lys Gly Cys Phe Pro Glu Lys Phe
370 375 380
Asp Leu Phe Lys Glu Val Met Ile Arg Asn Leu Arg Leu Cys Gly Arg
385 390 395 400
Glu Gly Lys Arg Lys Gly Val Asp Tyr Gly Lys Phe Ala Glu Glu Leu
405 410 415
Phe Leu Gln Leu Glu Lys Val Thr Leu Pro Ser Val Gly Asp Gly Pro
420 425 430
Trp Asn Asn Leu Arg Ser Gln Ser Lys Val Ser Leu Pro Leu Asp Gly
435 440 445
Ser Gly Asp Gly Pro Gln Ser Glu Phe Glu Ala Pro Ser Val Ser Gly
450 455 460
Ile Ser Gly Ser His Lys Lys Arg Arg Ile
465 470
<210> 3
<211> 3522
<212> DNA
<213> 人工序列()
<400> 3
atgtctaacg gtgacggtct gatccgttct ctggttgacg gtgacctgga aggtttccgt 60
cagggtttcg aatctttcct ggaccagtgc ccgtctttcc tgtaccacgt ttctgcgggt 120
cgtttcctgc cggttttctt cttctctatg ttctctaccg cgcacgacgc gaacatcctg 180
aacgcgaacg aacgtgttta cttccgtttc gacaaccacg gtgttaaccc gcgtaacggt 240
gaaaaccgta acaccgcgaa cctgaaagtt gcggtttacc gtgacggtca gcaggttgtt 300
cgttgctact ctatctctga ccgtccgaac tctgacggtc tgcgtttctc tacccgtgaa 360
cgtaacgcgc tggttcagga aatccgtcgt cagaacccga acctgcgtga agaagacctg 420
aacttcgaac agtacaaagt ttgcatgcac ggtaaaggta aatctcaggg tgaagcgatc 480
gcgaccgttt tcgaagttat ccgtgagaag gaccgtcagg gtcgtgacaa attcgcgaaa 540
tactctgcgt ctgaagttca cttcctgcgt cagctgttcc gtaaccaccg tctgaccatc 600
aaagaaatcg aaggtcgtca gctgaaccag aaccagctgc gtcagctggg tcgttctgtt 660
aacttcaccc gtgttgaacc gggtcagcag ggtatcgaca acttcatgga aatgctggcg 720
tctaaccaga aaattaacgt tgaggactct ctgcgtggtg acatcctgga atacgttacc 780
cgtacctaca acaactaccg tgcgcagatc gaaaacaaca tcgaaggtcg ttctcagaaa 840
ttcgaatctc acggtttcct gctgggtttc ctggcgaact tctctcaccg ttacaccatc 900
ggtgttgacc tggacctgtc tccgcgtaac tctcacgttg cgttcctggt tcgtcaccag 960
gttgaacgtg aaaacatccc gatcgttatc aacctggcga cccgtgcgcc gccgtacatc 1020
gcgctgaacc gtgcgcgttc tcacgcgaaa cgtctgcacg ttttctcttt catcccgatc 1080
cacaccgaat ctcgtaacac cgtttgcgtt ggtctgaact tcaacctgaa cctggacccg 1140
ttctctgttg acaccgttgg tctgcagcag gaccgtttcc cgctggttca gcgtctgttc 1200
gaatgcctgg aaaacgaagg tatccgtgaa aacatccgtg acttcctgct gcaccacctg 1260
ccggcggaaa tcccgcgtaa cgcggaaaac tacgaccgta tcttcgactg catcaccggt 1320
ttcgcgttcg gtaactctgc gttcgacgaa cacccgctgg ttaatgaaga agaagttcag 1380
gcgccgatca ccaaatacat cttccgtcac ggtgacgaag gtctgcgttg cctgaccatg 1440
gttttccacg cggaaggttc tgacatcgtt atcctgcaca tccgtgcgca cgacgcgcag 1500
cagcagggtg cgatcaacct gcagaccctg aacgttaacg gtaacgacgt tcacgtttgg 1560
gaagtttctt gcaccctgaa caaccagctg gaactggaca tcgacctgcc gaacgacctg 1620
ggtctgtacc acgactacca gaacaacaac gcgaacaact tcctggcggg tgacctggtt 1680
caggttccga acaccgaaaa cgttcacaac accctgaacc aggttgttaa cgacggttgg 1740
aagaatatcg cgcagcaccg tggtctgttc caggaaatct ctggtgcgct gatgccgctg 1800
gttgacacca tcaacgttaa ctctgaagac aaattccgtt ctatcctgca cggtaccttc 1860
tacgcgtctg acaacccgta caaagttctg gcgatgtaca aagttggtca gacctactct 1920
ctgaaacgtg gtcaggaaga agaaggtgaa cgtgttatcc tgacccgtat caccgaacag 1980
cgtctggacc tgctgctgct gcgtcagccg cgtgaaaacg acctggacac ccacccgatc 2040
ggttacgttc tgcgtctggc gaacaacgcg gaagaagttg gtcagcagca gaacgacgcg 2100
cgtcaggaaa tcggtcgtct gaagaagcag caccgtggtt tcatcccgat cacctctggt 2160
aacgaagttg ttctgttccc gatcgttttc aaccgtgacg cgcacgaagc gggtaacctg 2220
atcctgttcc cggaaggtat cggtcgtgaa gaacacgttc accgtctgga ccgtcacgtt 2280
cgttcttctc gtccgggtgg tctggttggt ccggaatctg ttatcgacga aaacccgccg 2340
gaaggtctgc tgtctgacca gacccgtgaa aacttccgtc gtttctacga agagaaggcg 2400
ccgggtcaga actctatctt cctgctggac atcggtgaca acctgcacgt tccgttctct 2460
tacctgcagg gtacccgtgc gcaggttatc gaaaccctga aatctcgtat ccgtggtggt 2520
ggtaccccga ccgcgcaggg tatcctgcag cagatcaacg cgatcctgcg tcgtaacaac 2580
gcgcgtgaaa tcgaagacgt tcacgacctg ctggcgctgg acttcgcgac cgacaaccag 2640
aactaccgtt actggctgca gacccacgac atgttcttcg cggcgcgtca gtacaccttc 2700
ctggacaacc agtctcactc taccaacgac cactacggtt tcgaaatcac ctctgttggt 2760
gttgacggta accagaacga cccgaccggt cgtggtctgc tgtcttctca catcaccaac 2820
ttcaaacaga aagttgactc tggtgagaag gaccgtctga tcgcgatcat caacgttggt 2880
aaccgtcact gggttaccct ggttatcgtt caccagaacg gtaactacta cggttactac 2940
gcggactctc tgggtccgga ctctggtatc gacaacaaca tccgtggtgc gctgcgtgaa 3000
tgcgacatca acgacgacaa cgttcacaac atctctgttc accagcagac cgacggtcac 3060
aactgcggta tctgggttta cgaaaacgcg cgtgacatca accaggcgat cgaccaggcg 3120
ctgcagggta acaacaactt cggtgagaag ggtgaaggta tcatcggtta catccgtggt 3180
ctgctgtctg cgggtatcgg taacgacacc cgtcagccgc gtcgtaacga acagtacttc 3240
gaagaccgtc gtcgtgacat ctctcagctg ctgcagaacg acccgaacct gccgtctcgt 3300
cgttctgacc tgatccaggc gcacccgggt atccagcacg aaatcgaccc gctgctgctg 3360
cagttcctgg gtctgcagta cccgcagcgt ggtggtggtg gtgcgctgca gctgggtggt 3420
gaacgtgtta tctctatcga cttcggtaac ccgcagtctg cgctggacaa aatcgacggt 3480
gtttctcgtg tttacaacca ctctaactct cgtggttctc gt 3522
<210> 4
<211> 1174
<212> PRT
<213> 人工序列()
<400> 4
Met Ser Asn Gly Asp Gly Leu Ile Arg Ser Leu Val Asp Gly Asp Leu
1 5 10 15
Glu Gly Phe Arg Gln Gly Phe Glu Ser Phe Leu Asp Gln Cys Pro Ser
20 25 30
Phe Leu Tyr His Val Ser Ala Gly Arg Phe Leu Pro Val Phe Phe Phe
35 40 45
Ser Met Phe Ser Thr Ala His Asp Ala Asn Ile Leu Asn Ala Asn Glu
50 55 60
Arg Val Tyr Phe Arg Phe Asp Asn His Gly Val Asn Pro Arg Asn Gly
65 70 75 80
Glu Asn Arg Asn Thr Ala Asn Leu Lys Val Ala Val Tyr Arg Asp Gly
85 90 95
Gln Gln Val Val Arg Cys Tyr Ser Ile Ser Asp Arg Pro Asn Ser Asp
100 105 110
Gly Leu Arg Phe Ser Thr Arg Glu Arg Asn Ala Leu Val Gln Glu Ile
115 120 125
Arg Arg Gln Asn Pro Asn Leu Arg Glu Glu Asp Leu Asn Phe Glu Gln
130 135 140
Tyr Lys Val Cys Met His Gly Lys Gly Lys Ser Gln Gly Glu Ala Ile
145 150 155 160
Ala Thr Val Phe Glu Val Ile Arg Glu Lys Asp Arg Gln Gly Arg Asp
165 170 175
Lys Phe Ala Lys Tyr Ser Ala Ser Glu Val His Phe Leu Arg Gln Leu
180 185 190
Phe Arg Asn His Arg Leu Thr Ile Lys Glu Ile Glu Gly Arg Gln Leu
195 200 205
Asn Gln Asn Gln Leu Arg Gln Leu Gly Arg Ser Val Asn Phe Thr Arg
210 215 220
Val Glu Pro Gly Gln Gln Gly Ile Asp Asn Phe Met Glu Met Leu Ala
225 230 235 240
Ser Asn Gln Lys Ile Asn Val Glu Asp Ser Leu Arg Gly Asp Ile Leu
245 250 255
Glu Tyr Val Thr Arg Thr Tyr Asn Asn Tyr Arg Ala Gln Ile Glu Asn
260 265 270
Asn Ile Glu Gly Arg Ser Gln Lys Phe Glu Ser His Gly Phe Leu Leu
275 280 285
Gly Phe Leu Ala Asn Phe Ser His Arg Tyr Thr Ile Gly Val Asp Leu
290 295 300
Asp Leu Ser Pro Arg Asn Ser His Val Ala Phe Leu Val Arg His Gln
305 310 315 320
Val Glu Arg Glu Asn Ile Pro Ile Val Ile Asn Leu Ala Thr Arg Ala
325 330 335
Pro Pro Tyr Ile Ala Leu Asn Arg Ala Arg Ser His Ala Lys Arg Leu
340 345 350
His Val Phe Ser Phe Ile Pro Ile His Thr Glu Ser Arg Asn Thr Val
355 360 365
Cys Val Gly Leu Asn Phe Asn Leu Asn Leu Asp Pro Phe Ser Val Asp
370 375 380
Thr Val Gly Leu Gln Gln Asp Arg Phe Pro Leu Val Gln Arg Leu Phe
385 390 395 400
Glu Cys Leu Glu Asn Glu Gly Ile Arg Glu Asn Ile Arg Asp Phe Leu
405 410 415
Leu His His Leu Pro Ala Glu Ile Pro Arg Asn Ala Glu Asn Tyr Asp
420 425 430
Arg Ile Phe Asp Cys Ile Thr Gly Phe Ala Phe Gly Asn Ser Ala Phe
435 440 445
Asp Glu His Pro Leu Val Asn Glu Glu Glu Val Gln Ala Pro Ile Thr
450 455 460
Lys Tyr Ile Phe Arg His Gly Asp Glu Gly Leu Arg Cys Leu Thr Met
465 470 475 480
Val Phe His Ala Glu Gly Ser Asp Ile Val Ile Leu His Ile Arg Ala
485 490 495
His Asp Ala Gln Gln Gln Gly Ala Ile Asn Leu Gln Thr Leu Asn Val
500 505 510
Asn Gly Asn Asp Val His Val Trp Glu Val Ser Cys Thr Leu Asn Asn
515 520 525
Gln Leu Glu Leu Asp Ile Asp Leu Pro Asn Asp Leu Gly Leu Tyr His
530 535 540
Asp Tyr Gln Asn Asn Asn Ala Asn Asn Phe Leu Ala Gly Asp Leu Val
545 550 555 560
Gln Val Pro Asn Thr Glu Asn Val His Asn Thr Leu Asn Gln Val Val
565 570 575
Asn Asp Gly Trp Lys Asn Ile Ala Gln His Arg Gly Leu Phe Gln Glu
580 585 590
Ile Ser Gly Ala Leu Met Pro Leu Val Asp Thr Ile Asn Val Asn Ser
595 600 605
Glu Asp Lys Phe Arg Ser Ile Leu His Gly Thr Phe Tyr Ala Ser Asp
610 615 620
Asn Pro Tyr Lys Val Leu Ala Met Tyr Lys Val Gly Gln Thr Tyr Ser
625 630 635 640
Leu Lys Arg Gly Gln Glu Glu Glu Gly Glu Arg Val Ile Leu Thr Arg
645 650 655
Ile Thr Glu Gln Arg Leu Asp Leu Leu Leu Leu Arg Gln Pro Arg Glu
660 665 670
Asn Asp Leu Asp Thr His Pro Ile Gly Tyr Val Leu Arg Leu Ala Asn
675 680 685
Asn Ala Glu Glu Val Gly Gln Gln Gln Asn Asp Ala Arg Gln Glu Ile
690 695 700
Gly Arg Leu Lys Lys Gln His Arg Gly Phe Ile Pro Ile Thr Ser Gly
705 710 715 720
Asn Glu Val Val Leu Phe Pro Ile Val Phe Asn Arg Asp Ala His Glu
725 730 735
Ala Gly Asn Leu Ile Leu Phe Pro Glu Gly Ile Gly Arg Glu Glu His
740 745 750
Val His Arg Leu Asp Arg His Val Arg Ser Ser Arg Pro Gly Gly Leu
755 760 765
Val Gly Pro Glu Ser Val Ile Asp Glu Asn Pro Pro Glu Gly Leu Leu
770 775 780
Ser Asp Gln Thr Arg Glu Asn Phe Arg Arg Phe Tyr Glu Glu Lys Ala
785 790 795 800
Pro Gly Gln Asn Ser Ile Phe Leu Leu Asp Ile Gly Asp Asn Leu His
805 810 815
Val Pro Phe Ser Tyr Leu Gln Gly Thr Arg Ala Gln Val Ile Glu Thr
820 825 830
Leu Lys Ser Arg Ile Arg Gly Gly Gly Thr Pro Thr Ala Gln Gly Ile
835 840 845
Leu Gln Gln Ile Asn Ala Ile Leu Arg Arg Asn Asn Ala Arg Glu Ile
850 855 860
Glu Asp Val His Asp Leu Leu Ala Leu Asp Phe Ala Thr Asp Asn Gln
865 870 875 880
Asn Tyr Arg Tyr Trp Leu Gln Thr His Asp Met Phe Phe Ala Ala Arg
885 890 895
Gln Tyr Thr Phe Leu Asp Asn Gln Ser His Ser Thr Asn Asp His Tyr
900 905 910
Gly Phe Glu Ile Thr Ser Val Gly Val Asp Gly Asn Gln Asn Asp Pro
915 920 925
Thr Gly Arg Gly Leu Leu Ser Ser His Ile Thr Asn Phe Lys Gln Lys
930 935 940
Val Asp Ser Gly Glu Lys Asp Arg Leu Ile Ala Ile Ile Asn Val Gly
945 950 955 960
Asn Arg His Trp Val Thr Leu Val Ile Val His Gln Asn Gly Asn Tyr
965 970 975
Tyr Gly Tyr Tyr Ala Asp Ser Leu Gly Pro Asp Ser Gly Ile Asp Asn
980 985 990
Asn Ile Arg Gly Ala Leu Arg Glu Cys Asp Ile Asn Asp Asp Asn Val
995 1000 1005
His Asn Ile Ser Val His Gln Gln Thr Asp Gly His Asn Cys Gly Ile
1010 1015 1020
Trp Val Tyr Glu Asn Ala Arg Asp Ile Asn Gln Ala Ile Asp Gln Ala
1025 1030 1035 1040
Leu Gln Gly Asn Asn Asn Phe Gly Glu Lys Gly Glu Gly Ile Ile Gly
1045 1050 1055
Tyr Ile Arg Gly Leu Leu Ser Ala Gly Ile Gly Asn Asp Thr Arg Gln
1060 1065 1070
Pro Arg Arg Asn Glu Gln Tyr Phe Glu Asp Arg Arg Arg Asp Ile Ser
1075 1080 1085
Gln Leu Leu Gln Asn Asp Pro Asn Leu Pro Ser Arg Arg Ser Asp Leu
1090 1095 1100
Ile Gln Ala His Pro Gly Ile Gln His Glu Ile Asp Pro Leu Leu Leu
1105 1110 1115 1120
Gln Phe Leu Gly Leu Gln Tyr Pro Gln Arg Gly Gly Gly Gly Ala Leu
1125 1130 1135
Gln Leu Gly Gly Glu Arg Val Ile Ser Ile Asp Phe Gly Asn Pro Gln
1140 1145 1150
Ser Ala Leu Asp Lys Ile Asp Gly Val Ser Arg Val Tyr Asn His Ser
1155 1160 1165
Asn Ser Arg Gly Ser Arg
1170
<210> 5
<211> 1335
<212> DNA
<213> 人工序列()
<400> 5
atggaatctg gtctggacca caactacaac aaaatcctgg acatcctgaa aggtgcgatc 60
aaaggtgacg acaaccaggt taaagcgcgt aaacacctgc gtgttgaacg ttggctgcgt 120
gcgtacatcc agctgatccg ccgcttcgac gaagaggaac tgatcttctt ctctgacatc 180
ttctctgaca actcttgctg ggacggtatc aaactgaaga ataaagcggt tggtgaacgt 240
ctgaccgaag agaagaacaa gaatggtaaa gaaaacccgc tggacctggc gcgtcgttac 300
tacctggcgt gcaaatactg cctggaagac aaaatcccgg gtctgttcga acaggttttc 360
atgcgtttca aacgttctgc gttcgaagaa gacggttctg acgacgacct gcgtcgtcgt 420
ctgctggaaa acatcgaaga aacctctccg atccgtgcgt tctggtcttt cctgatcgac 480
aaacagatcg gtaaactgaa cgaatacaaa tctgttgaag gtctgcagaa atctatccag 540
atcaactcta acgataattg ggaagaaggt atcgaattct tctacaacaa actgcacaac 600
gactcttcta tctcttctca ggacaaagac gacctgctga tcgaagcggc gctgtctgcg 660
gttaaaggtt acgaagaagt tgacaccatc gaattctgcc tgtctaaaat ggacgacgaa 720
cagaagaaga aactgctgga ccgtgactac aaagaaaaca cctactacgc ggttctgaac 780
gttctggttg gtcagtacta cttcgactct ttcatggaac tgtctcgtct gtgctctcag 840
atcgaatgcg aagaatacac caccttcctg tcttctctgt ctcgtcaggt tctggaaaat 900
ccggacctgt ctgaagaaac caagaagtgc atgatgaacg tttgggaacg tatcatcaaa 960
ctgaaaaccc aggaccgtgg tgaacagtct atctcttcta tcttcgttga ctactctgtt 1020
acctacacca tcgcgaacct gatcgttgac ccgtctcgtc agggtgtttc taaagaagaa 1080
atcctgggta aaatcctgaa acacgttaaa gaaatgtctg gtgaagaaat gatcaaagtt 1140
aaagactctg ttctgtctaa aatccagctg ttccacggtg gtaagaagct gcagctgggt 1200
gaacaggttt tctctaaact ggcgcaggaa gcgtctaaag aatctatcct gcgtgaagcg 1260
ggtgacaccc tgccgcagtc ttctctgtct accaccgaca ccccgtacaa catcaaatct 1320
ctgtctcact ctaaa 1335
<210> 6
<211> 445
<212> PRT
<213> 人工序列()
<400> 6
Met Glu Ser Gly Leu Asp His Asn Tyr Asn Lys Ile Leu Asp Ile Leu
1 5 10 15
Lys Gly Ala Ile Lys Gly Asp Asp Asn Gln Val Lys Ala Arg Lys His
20 25 30
Leu Arg Val Glu Arg Trp Leu Arg Ala Tyr Ile Gln Leu Ile Arg Arg
35 40 45
Phe Asp Glu Glu Glu Leu Ile Phe Phe Ser Asp Ile Phe Ser Asp Asn
50 55 60
Ser Cys Trp Asp Gly Ile Lys Leu Lys Asn Lys Ala Val Gly Glu Arg
65 70 75 80
Leu Thr Glu Glu Lys Asn Lys Asn Gly Lys Glu Asn Pro Leu Asp Leu
85 90 95
Ala Arg Arg Tyr Tyr Leu Ala Cys Lys Tyr Cys Leu Glu Asp Lys Ile
100 105 110
Pro Gly Leu Phe Glu Gln Val Phe Met Arg Phe Lys Arg Ser Ala Phe
115 120 125
Glu Glu Asp Gly Ser Asp Asp Asp Leu Arg Arg Arg Leu Leu Glu Asn
130 135 140
Ile Glu Glu Thr Ser Pro Ile Arg Ala Phe Trp Ser Phe Leu Ile Asp
145 150 155 160
Lys Gln Ile Gly Lys Leu Asn Glu Tyr Lys Ser Val Glu Gly Leu Gln
165 170 175
Lys Ser Ile Gln Ile Asn Ser Asn Asp Asn Trp Glu Glu Gly Ile Glu
180 185 190
Phe Phe Tyr Asn Lys Leu His Asn Asp Ser Ser Ile Ser Ser Gln Asp
195 200 205
Lys Asp Asp Leu Leu Ile Glu Ala Ala Leu Ser Ala Val Lys Gly Tyr
210 215 220
Glu Glu Val Asp Thr Ile Glu Phe Cys Leu Ser Lys Met Asp Asp Glu
225 230 235 240
Gln Lys Lys Lys Leu Leu Asp Arg Asp Tyr Lys Glu Asn Thr Tyr Tyr
245 250 255
Ala Val Leu Asn Val Leu Val Gly Gln Tyr Tyr Phe Asp Ser Phe Met
260 265 270
Glu Leu Ser Arg Leu Cys Ser Gln Ile Glu Cys Glu Glu Tyr Thr Thr
275 280 285
Phe Leu Ser Ser Leu Ser Arg Gln Val Leu Glu Asn Pro Asp Leu Ser
290 295 300
Glu Glu Thr Lys Lys Cys Met Met Asn Val Trp Glu Arg Ile Ile Lys
305 310 315 320
Leu Lys Thr Gln Asp Arg Gly Glu Gln Ser Ile Ser Ser Ile Phe Val
325 330 335
Asp Tyr Ser Val Thr Tyr Thr Ile Ala Asn Leu Ile Val Asp Pro Ser
340 345 350
Arg Gln Gly Val Ser Lys Glu Glu Ile Leu Gly Lys Ile Leu Lys His
355 360 365
Val Lys Glu Met Ser Gly Glu Glu Met Ile Lys Val Lys Asp Ser Val
370 375 380
Leu Ser Lys Ile Gln Leu Phe His Gly Gly Lys Lys Leu Gln Leu Gly
385 390 395 400
Glu Gln Val Phe Ser Lys Leu Ala Gln Glu Ala Ser Lys Glu Ser Ile
405 410 415
Leu Arg Glu Ala Gly Asp Thr Leu Pro Gln Ser Ser Leu Ser Thr Thr
420 425 430
Asp Thr Pro Tyr Asn Ile Lys Ser Leu Ser His Ser Lys
435 440 445
<210> 7
<211> 2196
<212> DNA
<213> 人工序列()
<400> 7
atgccgtcta acgttaaacc gctggaactg gttcagctgc tgctgatgcg taacaaatct 60
aaagacgaat tcctggactt ccagaaacgt ttccagtctt tcatcaacca gtctccgtct 120
ttcctgcact ctgttggtaa accgggtttc ttcccgtctt tcttcttcgg tatgttcgcg 180
accgttctgg acaccgaact ggcgaccaaa atcggtatca agaagctgca cttccgtttc 240
gacgacaacc gtaccctgaa aatcgcgatc ctgaccaacg aaggtctgaa atgcatcacc 300
atgtctgacc aggttgacgg taacatgcac ctgaaattct ctcagggtga actggagaag 360
atcgcgcaga aatggaaaat gggtgcggaa ttcgacaaac tggagaagga agaacacgaa 420
atcaccatca ccggtaaaga agttaaacac ggtaaagttg acccggcgtt ctctaagaag 480
accgactact ctcagaaagg tttcaccgaa atcgagaagg accgtgacca gcaggacctg 540
gaatctctga tctctaaact gtctaaccag cgtttcgaag aagttaagaa gaacgcggaa 600
cgtatgttca actacatcac caacgtttac aagaagtacg agaaggaaac cctgttctct 660
ggtaaagaat cttctcacca cggtttcctg gcgggtttcc tgatcaactt caaataccgt 720
ttccacctga aactgtacct ggaactgttc gcgggtaaag gttacgcgga catcatcctg 780
ctggttcgtg gttctgacaa atctctgtct tctatcccga tcatcatcga actgaaagcg 840
ggtaccggtg aaatctctac cgttatcaaa gcgctgaaac aggcgcagga ctacgttaaa 900
ggttctttct ctaactctat ccgtatgatc accatcgcga accgtgcgat ctgcgttggt 960
ctgaacttcg acatggttca ccacgaaaac gttaaaatcc gtgttgaaaa cttcctgtct 1020
cgtcgtggta actctgttat cgaggaactg ctgggtaccc gtgcgaccaa cgcggaagtt 1080
atccgtaccc agctggaata cctgtactac ggtatcgttt ggtctaacgg tggttctgac 1140
aacatcaact acgtttctcg tatgatcctg ggtcagctgg ttctgatctc taacatcatc 1200
gatgaagagg aactgggtaa acacatcttc atctacgacc agaacgacaa aatggttacc 1260
ggttctcaga aacgtccgga agcggcgaaa gaatctatcg aagactgcgt taccaccatc 1320
gttctgaccc tgggtaagaa ggttctgatc ctgaacatca acgagaagaa cgaattcgcg 1380
ctgcgtgttc cggacaacaa aggtatcccg atcgaaaaca tccgtcgtat ccagaacgtt 1440
aacgacatca aaatccagga aatcacctgc aacctgtact ctaccccgtc taacaagaat 1500
ccgttcgacc agtactgcaa caagaataaa ggtatcaccg ttaacaccta cgactctctg 1560
gacaaataca aacgtggtaa agaaatcctg cagggtaact tcacccgtat cgttgaaaac 1620
aagaagttca aagcggcgct gtctaaagcg atcgaatctg gtaaatacga cgactacaag 1680
aagctgttcg aagaaatctc tcacatcctg cacccgttca aatctctgat ctctaacgaa 1740
gcgaccttcc aggcggttct gcacggtctg ttctcttctt acggtgaaga caacatcaaa 1800
gttatcaccg aattccagat cggtggtggt gagaagctgg acgttatgct ggttatcaac 1860
gcgaccgacc agaagaagga atacccgccg gttggtatcg aactgaaatt cgcgaagaag 1920
ggtgaactgg acaagaagga gaaggacgcg aaagaccagc tgaaacgtta caaagaaggt 1980
gaagcgtaca aagttatcac cgacgcgggt aaagttaaac tgatctacgc ggttttcaac 2040
aaaggtgcga ccgacgaagg ttctctgatc aaaatcggta acgaattcgt tgaagttgac 2100
gttcgtcact cttctgttgt tgcgttcggt cagcagccgg gttctctgca gcagccgtac 2160
gttaaacagg cgggtctgtc tcgtgcggtt aaccag 2196
<210> 8
<211> 732
<212> PRT
<213> 人工序列()
<400> 8
Met Pro Ser Asn Val Lys Pro Leu Glu Leu Val Gln Leu Leu Leu Met
1 5 10 15
Arg Asn Lys Ser Lys Asp Glu Phe Leu Asp Phe Gln Lys Arg Phe Gln
20 25 30
Ser Phe Ile Asn Gln Ser Pro Ser Phe Leu His Ser Val Gly Lys Pro
35 40 45
Gly Phe Phe Pro Ser Phe Phe Phe Gly Met Phe Ala Thr Val Leu Asp
50 55 60
Thr Glu Leu Ala Thr Lys Ile Gly Ile Lys Lys Leu His Phe Arg Phe
65 70 75 80
Asp Asp Asn Arg Thr Leu Lys Ile Ala Ile Leu Thr Asn Glu Gly Leu
85 90 95
Lys Cys Ile Thr Met Ser Asp Gln Val Asp Gly Asn Met His Leu Lys
100 105 110
Phe Ser Gln Gly Glu Leu Glu Lys Ile Ala Gln Lys Trp Lys Met Gly
115 120 125
Ala Glu Phe Asp Lys Leu Glu Lys Glu Glu His Glu Ile Thr Ile Thr
130 135 140
Gly Lys Glu Val Lys His Gly Lys Val Asp Pro Ala Phe Ser Lys Lys
145 150 155 160
Thr Asp Tyr Ser Gln Lys Gly Phe Thr Glu Ile Glu Lys Asp Arg Asp
165 170 175
Gln Gln Asp Leu Glu Ser Leu Ile Ser Lys Leu Ser Asn Gln Arg Phe
180 185 190
Glu Glu Val Lys Lys Asn Ala Glu Arg Met Phe Asn Tyr Ile Thr Asn
195 200 205
Val Tyr Lys Lys Tyr Glu Lys Glu Thr Leu Phe Ser Gly Lys Glu Ser
210 215 220
Ser His His Gly Phe Leu Ala Gly Phe Leu Ile Asn Phe Lys Tyr Arg
225 230 235 240
Phe His Leu Lys Leu Tyr Leu Glu Leu Phe Ala Gly Lys Gly Tyr Ala
245 250 255
Asp Ile Ile Leu Leu Val Arg Gly Ser Asp Lys Ser Leu Ser Ser Ile
260 265 270
Pro Ile Ile Ile Glu Leu Lys Ala Gly Thr Gly Glu Ile Ser Thr Val
275 280 285
Ile Lys Ala Leu Lys Gln Ala Gln Asp Tyr Val Lys Gly Ser Phe Ser
290 295 300
Asn Ser Ile Arg Met Ile Thr Ile Ala Asn Arg Ala Ile Cys Val Gly
305 310 315 320
Leu Asn Phe Asp Met Val His His Glu Asn Val Lys Ile Arg Val Glu
325 330 335
Asn Phe Leu Ser Arg Arg Gly Asn Ser Val Ile Glu Glu Leu Leu Gly
340 345 350
Thr Arg Ala Thr Asn Ala Glu Val Ile Arg Thr Gln Leu Glu Tyr Leu
355 360 365
Tyr Tyr Gly Ile Val Trp Ser Asn Gly Gly Ser Asp Asn Ile Asn Tyr
370 375 380
Val Ser Arg Met Ile Leu Gly Gln Leu Val Leu Ile Ser Asn Ile Ile
385 390 395 400
Asp Glu Glu Glu Leu Gly Lys His Ile Phe Ile Tyr Asp Gln Asn Asp
405 410 415
Lys Met Val Thr Gly Ser Gln Lys Arg Pro Glu Ala Ala Lys Glu Ser
420 425 430
Ile Glu Asp Cys Val Thr Thr Ile Val Leu Thr Leu Gly Lys Lys Val
435 440 445
Leu Ile Leu Asn Ile Asn Glu Lys Asn Glu Phe Ala Leu Arg Val Pro
450 455 460
Asp Asn Lys Gly Ile Pro Ile Glu Asn Ile Arg Arg Ile Gln Asn Val
465 470 475 480
Asn Asp Ile Lys Ile Gln Glu Ile Thr Cys Asn Leu Tyr Ser Thr Pro
485 490 495
Ser Asn Lys Asn Pro Phe Asp Gln Tyr Cys Asn Lys Asn Lys Gly Ile
500 505 510
Thr Val Asn Thr Tyr Asp Ser Leu Asp Lys Tyr Lys Arg Gly Lys Glu
515 520 525
Ile Leu Gln Gly Asn Phe Thr Arg Ile Val Glu Asn Lys Lys Phe Lys
530 535 540
Ala Ala Leu Ser Lys Ala Ile Glu Ser Gly Lys Tyr Asp Asp Tyr Lys
545 550 555 560
Lys Leu Phe Glu Glu Ile Ser His Ile Leu His Pro Phe Lys Ser Leu
565 570 575
Ile Ser Asn Glu Ala Thr Phe Gln Ala Val Leu His Gly Leu Phe Ser
580 585 590
Ser Tyr Gly Glu Asp Asn Ile Lys Val Ile Thr Glu Phe Gln Ile Gly
595 600 605
Gly Gly Glu Lys Leu Asp Val Met Leu Val Ile Asn Ala Thr Asp Gln
610 615 620
Lys Lys Glu Tyr Pro Pro Val Gly Ile Glu Leu Lys Phe Ala Lys Lys
625 630 635 640
Gly Glu Leu Asp Lys Lys Glu Lys Asp Ala Lys Asp Gln Leu Lys Arg
645 650 655
Tyr Lys Glu Gly Glu Ala Tyr Lys Val Ile Thr Asp Ala Gly Lys Val
660 665 670
Lys Leu Ile Tyr Ala Val Phe Asn Lys Gly Ala Thr Asp Glu Gly Ser
675 680 685
Leu Ile Lys Ile Gly Asn Glu Phe Val Glu Val Asp Val Arg His Ser
690 695 700
Ser Val Val Ala Phe Gly Gln Gln Pro Gly Ser Leu Gln Gln Pro Tyr
705 710 715 720
Val Lys Gln Ala Gly Leu Ser Arg Ala Val Asn Gln
725 730
<210> 9
<211> 1422
<212> DNA
<213> 人工序列()
<400> 9
atgccgatcg aaaccaaacg tcaggcggaa gttctgaaga agctgcagga cgttatcaaa 60
cacaccgacc gtgacatcgc ggcgggtcgt aaactggcga tcaaacgttg ggttgaaacc 120
tacatcgaat acatcaaact gttcaaagac gacaaactgg aattcctgta caacgttttc 180
cgtgacgaag gttgctggct gggtacccgt ctgaacaaca ccgttctggg tcagaaactg 240
accgaagaga agatcggtga aatcgacaac ccgctgccgc gttacggtat ggcgtctcgt 300
tactgcatca ccggtaaaat cggtgacttc ttcaacaaac agttcgttct gtctcgtggt 360
cagttcacct ctgaagaagt tgactctcag ggtaacccga tctctgacca gtacgttcgt 420
aacatcctgc tgtcttctat gaaacgtaac ggtccggttt tcgacttctg gatcgaccgt 480
gaatctggtg aactgaagaa gtacgacgcg gttgaaggtt tcgactctac cgttaaactg 540
aaatggtctg aaggtgttga atacttctac aaccagctgg aagagaagga caaagagaag 600
aaactgaccg aagcgatcgt tgcgctgtct cgtccgcagt ctgttaaacg tgacgcgccg 660
atcctggact tctgcgttcg taacatcggt gacaaagaca ccctgctgca gaaactgctg 720
cagaaagaca aaggtgttta cttcctgctg gcggaactga tcgaatcttg cttcttcgac 780
accgttcacg acctggttca gtgctggtgc tacaaaggtg tttctgcggg tggtgactgc 840
tctgacaaaa tcttctctca gcaggactac gaactgttcc tgtactctct gtctaacgtt 900
atgctgaaga atccggaact gtctgttcag gcgcgttctc tgatcatgga aatctggaaa 960
tgcgaacgtt tcgcggaata ccgtgaaacc tctgttaaca cctctaacta caccgttccg 1020
atcaaatctg ttctgggtgg tctgatcatc aactggaaac gtgaagacgt ttgcaaaccg 1080
gaccgtgaaa tcgagaagga agaaatcctg gacatgatct ctttcgcgaa aggttgcttc 1140
ccggagaagt tcgacctgtt caaagaagtt atgatcgaaa acctgcgtat ctgcggtcgt 1200
gaaggtaaac gtaaaggtgt tgactacggt aaattcgcgg aagaactgtt cctgcagctg 1260
gagaaggtta ccctgccgtc tgttggtgac ggtccgtgga acaacctgcg ttctcagtct 1320
aaagtttctc tgccgctgga cggttctggt gacggtccgc agtctgaatt cgaagcgccg 1380
tctgtttctg gtatctctgg ttctcacaag aagcgtcgta tc 1422
<210> 10
<211> 1473
<212> DNA
<213> 人工序列()
<400> 10
atgccgaccc agaaagaact gcgtgacacc atgtctaaga agctgcagga agcgatcaaa 60
cacccggacc cggcggttgt tgcgggtcgt aaatctgcga tcaaacgttg ggttggtgtt 120
ctgcaggaca acttcatgga acacatcaaa tacttcaaag gtgacaaact gaaattcctg 180
cacaacgttt tccaggacga aggttgctgg tctggtgttc gtctggacaa cgcggcgctg 240
ggtcagcgtt tcaccgaaga gaagatcggt ggtatcgaca acccgctgcg taaatacgaa 300
atggcgtgct cttactgcgt tgttgacaaa atccacccgc tgttccagaa acgtttcgaa 360
tcttaccgta acaaattccc gccgggtgcg ttcgacggta aaaccgaaac cgaattcggt 420
aaatacgttc gtaactctct gctggactct atcaaacgta aaggtccggt tttcgacttc 480
tggatcgacc gtgaatctgg tgaactgaag aagtacgacg cggttgaagg tttcgactct 540
gcggttaaat tcaaatggtc tgaaggtgtt gaatacttct acaaccacct gaaagaagaa 600
gacaaagaga agaaactgac cgaagcgatc ctggcgctgt ctcgtgttca gtctgttgag 660
aaggacgcgc cgatcctgga cttctgcgtt aacaaaatcg ttgacaaaga caccctgctg 720
cagaaactgt ctcagaaaga caaaggtgtt tactctctgt tcgcggaact gatcgaatct 780
tgcttcttcg acaccgttca cgacctggtt cagtgctggt gctacaaaga agtttctgcg 840
ggtggtgacc actctgagaa gatcttctct cagcgtgact acgaactgtt cctgtcttct 900
ctgtctgaca ccatgctgaa gaatccggaa ctgtctgttc aggcgcgttc tctgatcatg 960
gaattctggg aatgcggttc tctgtaccag taccgtaaag cggcggttaa cacctctaac 1020
tacaccgttc cgacctctgg tgttttcgcg gaactgatcg ttaactggcg tcgtgaagac 1080
atctacaaaa ccgacgaaga gaaggaaatc gagaagaaag aaatcctgga catgatgtct 1140
ttcgcgaaag actgcttccc ggagaagttc gaactgttca agaagctgat catccgtgac 1200
ctgcgtctgt gcggtcgtga aggtaaacgt gttaacgttg actacggtct gttcgcggaa 1260
gaactgttct ctgaactgga gaagaccatc ctgccgccgg gtccggttgg tgacggtccg 1320
tgctctaacc tgcgttctcg ttctaaagcg cacggttcta agaagaccac cctgccggtt 1380
gacgactctc cgcagtctga actgggtacc ccgtctgttt ctggtgtttc ttcttacaag 1440
aagaaatctg ttttcaccct gtctggtaac aaa 1473
<210> 11
<211> 3522
<212> DNA
<213> 人工序列()
<400> 11
atgtctaacg gtgacggtct gatccgttct ctggttgacg gtgacctgga aggtttccgt 60
cagggtttcg aatctttcct ggaccagtgc ccgtctttcc tgtaccacgt ttctgcgggt 120
cgtttcctgc cggttttctt cttctctatg ttctctaccg cgcacgacgc gaacatcctg 180
aacgcgaacg aacgtgttta cttccgtttc gacaaccacg gtgttaaccc gcgtaacggt 240
gaaaaccgta acaccgcgaa cctgaaagtt gcggtttacc gtgacggtca gcaggttgtt 300
cgttgctact ctatctctga ccgtccgaac tctgacggtc tgcgtttctc tacccgtgaa 360
cgtaacgcgc tggttcagga aatccgtcgt cagaacccga acctgcgtga agaagacctg 420
aacttcgaac agtacaaagt ttgcatgcac ggtaaaggta aatctcaggg tgaagcgatc 480
gcgaccgttt tcgaagttat ccgtgagaag gaccgtcagg gtcgtgacaa attcgcgaaa 540
tactctgcgt ctgaagttca cttcctgcgt cagctgttcc gtaaccaccg tctgaccatc 600
aaagaaatcg aaggtcgtca gctgaaccag aaccagctgc gtcagctggg tcgttctgtt 660
aacttcaccc gtgttgaacc gggtcagcag cgtatcgaca acttcatgga aatgctggcg 720
tctaaccagc gtcaggacgt tcgtgactct ctgcgtggtg acatcctgga atacgttacc 780
gacacctaca acaactaccg tgcgcagatc gaaaacaaca tcgaaggtcg ttctcagaaa 840
ttcgaatctc acggtttcct gctgggtttc ctggcgaact tctctcaccg ttacaccatc 900
ggtgttgacc tggacctgtc tccgcgtaac tctcacgttg cgttcctggt tcgtcaccag 960
gttgaacgtg aaaacatccc gatcgttatc aacctggcga cccgtgcgcc gccgtacatc 1020
gcgctgaacc gtgcgcgttc tcacgcggaa cgtctgcacg ttttctcttt catcccgatc 1080
cacaccgaat ctcgtaacac cgtttgcgtt ggtctgaact tcaacctgaa cctggacccg 1140
ttctctgttg acaccgttgg tctgcagcag gaccgtttcc cgctggttca gcgtctgttc 1200
gaatgcctgg aaaacgaagg tatccgtgaa aacatccgtg acttcctgct gcaccacctg 1260
ccggcggaaa tcccgcgtaa cgcggaaaac tacgaccgta tcttcgactg catcaccggt 1320
ttcgcgttcg gtaactctgc gttcgaccgt cacccgctgg aactggaaga agaagacgaa 1380
gcgccgatca ccaaatacat cttccgtcac ggtgacgaag gtctgcgttg cctgaccatg 1440
gttttccacg cggaaggttc tgacatcgtt atcctgcaca tccgtgcgca cgacgcgcag 1500
cagcagggtg cgatcaacct gcagaccctg aacgttaacg gtaacgacgt tcacgtttgg 1560
gaagtttctt gcaccctgaa caaccagctg gaactggaca tcgacctgcc gaacgacctg 1620
ggtctgtacc acgactacca gaacaacaac gcgaacaact tcctggcggg tgacctggtt 1680
caggttccga acaccgaaaa cgttcacaac accctgaacc aggttgttaa cgacggttgg 1740
aagaatatcg cgcagcaccg tggtctgttc caggaaatct ctggtgcgct gatgccgctg 1800
gttgacacca tcaacgttaa ctctgaagac aaattccgtt ctatcctgca cggtaccttc 1860
tacgcgtctg acaacccgta caaagttctg gcgatgtaca aagttggtca gacctactct 1920
ctgaaacgtg gtcaggaaga agaaggtgaa cgtgttatcc tgacccgtat caccgaacag 1980
cgtctggacc tgctgctgct gcgtcagccg cgtgaaaacg acctggacac ccacccgatc 2040
ggttacgttc tgcgtctggc gaacaacgcg gaagaagttg gtcagcagca gaacgacgcg 2100
cgtcaggaaa tcggtcgtct gaagaagcag caccgtggtt tcatcccgat cacctctggt 2160
aacgaagttg ttctgttccc gatcgttttc aaccgtgacg cgcacgaagc gggtaacctg 2220
atcctgttcc cggaaggtat cggtcgtgaa gaacacgttc accgtctgga ccgtcacgtt 2280
cgttcttctc gtccgggtgg tctggttggt ccggaatctg ttatcgacga aaacccgccg 2340
gaaggtctgc tgtctgacca gacccgtgaa aacttccgtc gtttctacga agagaaggcg 2400
ccgggtcaga actctatctt cctgctggac atcggtgaca acctgcacgt tccgttctct 2460
tacctgcagg gtacccgtgc gcaggttatc gaaaccctga aatctcgtat ccgtggtggt 2520
ggtaccccga ccgcgcaggg tatcctgcag cagatcaacg cgatcctgcg tcgtaacaac 2580
gcgcgtgaaa tcgaagacgt tcacgacctg ctggcgctgg acttcgcgac cgacaaccag 2640
aactaccgtt actggctgca gacccacgac atgttcttcg cggcgcgtca gtacaccttc 2700
ctggacaacc agtctcactc taccaacgac cactacggtt tcgaaatcac ctctgttggt 2760
gttgacggta accagaacga cccgaccggt cgtggtctgc tgtcttctca catcaccaac 2820
ttcaaacaga aagttgactc tggtgagaag gaccgtctga tcgcgatcat caacgttggt 2880
aaccgtcact gggttaccct ggttatcgtt caccagaacg gtaactacta cggttactac 2940
gcggactctc tgggtccgga ctctggtatc gacaacaaca tccgtggtgc gctgcgtgaa 3000
tgcgacatca acgacgacaa cgttcacaac atctctgttc accagcagac cgacggtcac 3060
aactgcggta tctgggttta cgaaaacgcg cgtgacatca accaggcgat cgaccaggcg 3120
ctgcagggta acaacaactt cggtgagaag ggtgaaggta tcatcggtta catccgtggt 3180
ctgctgtctg cgggtatcgg taacgacacc cgtcagccgc gtcgtaacga acagtacttc 3240
gaagaccgtc gtcgtgacat ctctcagctg ctgcagaacg acccgaacct gccgtctcgt 3300
cgttctgacc tgatccaggc gcacccgggt atccagcacg aaatcgaccc gctgctgctg 3360
cagttcctgg gtctgcagta cccgcagcgt ggtggtggtg gtgcgctgca gctgggtggt 3420
gaacgtgtta tctctatcga cttcggtaac ccgcagtctg cgctggacaa aatcgacggt 3480
gtttctcgtg tttacaacca ctctaactct cgtggttctc gt 3522
<210> 12
<211> 1174
<212> PRT
<213> 人工序列()
<400> 12
Met Ser Asn Gly Asp Gly Leu Ile Arg Ser Leu Val Asp Gly Asp Leu
1 5 10 15
Glu Gly Phe Arg Gln Gly Phe Glu Ser Phe Leu Asp Gln Cys Pro Ser
20 25 30
Phe Leu Tyr His Val Ser Ala Gly Arg Phe Leu Pro Val Phe Phe Phe
35 40 45
Ser Met Phe Ser Thr Ala His Asp Ala Asn Ile Leu Asn Ala Asn Glu
50 55 60
Arg Val Tyr Phe Arg Phe Asp Asn His Gly Val Asn Pro Arg Asn Gly
65 70 75 80
Glu Asn Arg Asn Thr Ala Asn Leu Lys Val Ala Val Tyr Arg Asp Gly
85 90 95
Gln Gln Val Val Arg Cys Tyr Ser Ile Ser Asp Arg Pro Asn Ser Asp
100 105 110
Gly Leu Arg Phe Ser Thr Arg Glu Arg Asn Ala Leu Val Gln Glu Ile
115 120 125
Arg Arg Gln Asn Pro Asn Leu Arg Glu Glu Asp Leu Asn Phe Glu Gln
130 135 140
Tyr Lys Val Cys Met His Gly Lys Gly Lys Ser Gln Gly Glu Ala Ile
145 150 155 160
Ala Thr Val Phe Glu Val Ile Arg Glu Lys Asp Arg Gln Gly Arg Asp
165 170 175
Lys Phe Ala Lys Tyr Ser Ala Ser Glu Val His Phe Leu Arg Gln Leu
180 185 190
Phe Arg Asn His Arg Leu Thr Ile Lys Glu Ile Glu Gly Arg Gln Leu
195 200 205
Asn Gln Asn Gln Leu Arg Gln Leu Gly Arg Ser Val Asn Phe Thr Arg
210 215 220
Val Glu Pro Gly Gln Gln Arg Ile Asp Asn Phe Met Glu Met Leu Ala
225 230 235 240
Ser Asn Gln Arg Gln Asp Val Arg Asp Ser Leu Arg Gly Asp Ile Leu
245 250 255
Glu Tyr Val Thr Asp Thr Tyr Asn Asn Tyr Arg Ala Gln Ile Glu Asn
260 265 270
Asn Ile Glu Gly Arg Ser Gln Lys Phe Glu Ser His Gly Phe Leu Leu
275 280 285
Gly Phe Leu Ala Asn Phe Ser His Arg Tyr Thr Ile Gly Val Asp Leu
290 295 300
Asp Leu Ser Pro Arg Asn Ser His Val Ala Phe Leu Val Arg His Gln
305 310 315 320
Val Glu Arg Glu Asn Ile Pro Ile Val Ile Asn Leu Ala Thr Arg Ala
325 330 335
Pro Pro Tyr Ile Ala Leu Asn Arg Ala Arg Ser His Ala Glu Arg Leu
340 345 350
His Val Phe Ser Phe Ile Pro Ile His Thr Glu Ser Arg Asn Thr Val
355 360 365
Cys Val Gly Leu Asn Phe Asn Leu Asn Leu Asp Pro Phe Ser Val Asp
370 375 380
Thr Val Gly Leu Gln Gln Asp Arg Phe Pro Leu Val Gln Arg Leu Phe
385 390 395 400
Glu Cys Leu Glu Asn Glu Gly Ile Arg Glu Asn Ile Arg Asp Phe Leu
405 410 415
Leu His His Leu Pro Ala Glu Ile Pro Arg Asn Ala Glu Asn Tyr Asp
420 425 430
Arg Ile Phe Asp Cys Ile Thr Gly Phe Ala Phe Gly Asn Ser Ala Phe
435 440 445
Asp Arg His Pro Leu Glu Leu Glu Glu Glu Asp Glu Ala Pro Ile Thr
450 455 460
Lys Tyr Ile Phe Arg His Gly Asp Glu Gly Leu Arg Cys Leu Thr Met
465 470 475 480
Val Phe His Ala Glu Gly Ser Asp Ile Val Ile Leu His Ile Arg Ala
485 490 495
His Asp Ala Gln Gln Gln Gly Ala Ile Asn Leu Gln Thr Leu Asn Val
500 505 510
Asn Gly Asn Asp Val His Val Trp Glu Val Ser Cys Thr Leu Asn Asn
515 520 525
Gln Leu Glu Leu Asp Ile Asp Leu Pro Asn Asp Leu Gly Leu Tyr His
530 535 540
Asp Tyr Gln Asn Asn Asn Ala Asn Asn Phe Leu Ala Gly Asp Leu Val
545 550 555 560
Gln Val Pro Asn Thr Glu Asn Val His Asn Thr Leu Asn Gln Val Val
565 570 575
Asn Asp Gly Trp Lys Asn Ile Ala Gln His Arg Gly Leu Phe Gln Glu
580 585 590
Ile Ser Gly Ala Leu Met Pro Leu Val Asp Thr Ile Asn Val Asn Ser
595 600 605
Glu Asp Lys Phe Arg Ser Ile Leu His Gly Thr Phe Tyr Ala Ser Asp
610 615 620
Asn Pro Tyr Lys Val Leu Ala Met Tyr Lys Val Gly Gln Thr Tyr Ser
625 630 635 640
Leu Lys Arg Gly Gln Glu Glu Glu Gly Glu Arg Val Ile Leu Thr Arg
645 650 655
Ile Thr Glu Gln Arg Leu Asp Leu Leu Leu Leu Arg Gln Pro Arg Glu
660 665 670
Asn Asp Leu Asp Thr His Pro Ile Gly Tyr Val Leu Arg Leu Ala Asn
675 680 685
Asn Ala Glu Glu Val Gly Gln Gln Gln Asn Asp Ala Arg Gln Glu Ile
690 695 700
Gly Arg Leu Lys Lys Gln His Arg Gly Phe Ile Pro Ile Thr Ser Gly
705 710 715 720
Asn Glu Val Val Leu Phe Pro Ile Val Phe Asn Arg Asp Ala His Glu
725 730 735
Ala Gly Asn Leu Ile Leu Phe Pro Glu Gly Ile Gly Arg Glu Glu His
740 745 750
Val His Arg Leu Asp Arg His Val Arg Ser Ser Arg Pro Gly Gly Leu
755 760 765
Val Gly Pro Glu Ser Val Ile Asp Glu Asn Pro Pro Glu Gly Leu Leu
770 775 780
Ser Asp Gln Thr Arg Glu Asn Phe Arg Arg Phe Tyr Glu Glu Lys Ala
785 790 795 800
Pro Gly Gln Asn Ser Ile Phe Leu Leu Asp Ile Gly Asp Asn Leu His
805 810 815
Val Pro Phe Ser Tyr Leu Gln Gly Thr Arg Ala Gln Val Ile Glu Thr
820 825 830
Leu Lys Ser Arg Ile Arg Gly Gly Gly Thr Pro Thr Ala Gln Gly Ile
835 840 845
Leu Gln Gln Ile Asn Ala Ile Leu Arg Arg Asn Asn Ala Arg Glu Ile
850 855 860
Glu Asp Val His Asp Leu Leu Ala Leu Asp Phe Ala Thr Asp Asn Gln
865 870 875 880
Asn Tyr Arg Tyr Trp Leu Gln Thr His Asp Met Phe Phe Ala Ala Arg
885 890 895
Gln Tyr Thr Phe Leu Asp Asn Gln Ser His Ser Thr Asn Asp His Tyr
900 905 910
Gly Phe Glu Ile Thr Ser Val Gly Val Asp Gly Asn Gln Asn Asp Pro
915 920 925
Thr Gly Arg Gly Leu Leu Ser Ser His Ile Thr Asn Phe Lys Gln Lys
930 935 940
Val Asp Ser Gly Glu Lys Asp Arg Leu Ile Ala Ile Ile Asn Val Gly
945 950 955 960
Asn Arg His Trp Val Thr Leu Val Ile Val His Gln Asn Gly Asn Tyr
965 970 975
Tyr Gly Tyr Tyr Ala Asp Ser Leu Gly Pro Asp Ser Gly Ile Asp Asn
980 985 990
Asn Ile Arg Gly Ala Leu Arg Glu Cys Asp Ile Asn Asp Asp Asn Val
995 1000 1005
His Asn Ile Ser Val His Gln Gln Thr Asp Gly His Asn Cys Gly Ile
1010 1015 1020
Trp Val Tyr Glu Asn Ala Arg Asp Ile Asn Gln Ala Ile Asp Gln Ala
1025 1030 1035 1040
Leu Gln Gly Asn Asn Asn Phe Gly Glu Lys Gly Glu Gly Ile Ile Gly
1045 1050 1055
Tyr Ile Arg Gly Leu Leu Ser Ala Gly Ile Gly Asn Asp Thr Arg Gln
1060 1065 1070
Pro Arg Arg Asn Glu Gln Tyr Phe Glu Asp Arg Arg Arg Asp Ile Ser
1075 1080 1085
Gln Leu Leu Gln Asn Asp Pro Asn Leu Pro Ser Arg Arg Ser Asp Leu
1090 1095 1100
Ile Gln Ala His Pro Gly Ile Gln His Glu Ile Asp Pro Leu Leu Leu
1105 1110 1115 1120
Gln Phe Leu Gly Leu Gln Tyr Pro Gln Arg Gly Gly Gly Gly Ala Leu
1125 1130 1135
Gln Leu Gly Gly Glu Arg Val Ile Ser Ile Asp Phe Gly Asn Pro Gln
1140 1145 1150
Ser Ala Leu Asp Lys Ile Asp Gly Val Ser Arg Val Tyr Asn His Ser
1155 1160 1165
Asn Ser Arg Gly Ser Arg
1170
<210> 13
<211> 34
<212> DNA
<213> 人工序列()
<400> 13
ggaattccat atgccaatcg aaaccaagaa acaa 34
<210> 14
<211> 31
<212> DNA
<213> 人工序列()
<400> 14
ccgctcgagg atgcggcgtt ttttatggct g 31
<210> 15
<211> 57
<212> DNA
<213> 人工序列()
<400> 15
cgggatccat ggactataaa gacgatgacg ataaaatgcc aatcgaaacc aagaaac 57
<210> 16
<211> 35
<212> DNA
<213> 人工序列()
<400> 16
acgcgtcgac ttagatgcgg cgttttttat ggctg 35
<210> 17
<211> 3498
<212> DNA
<213> 人工序列()
<400> 17
atggacggtg acctggacgg tttccgtcag gaattcgaat ctttcctgga ccagtgcccg 60
ttcttcctgt accacgtttc taccggtcgt ttcctgccgg ttttcttctt ctctatgttc 120
gcgaccgcgc acgacgcgaa catcctgaaa gcgaacgaac gtgtttactt ccgtttcgac 180
aaccacggta tcgacaccgg tggtcgtaac cgtaacaccg gtaacctgaa agttgcggtt 240
taccacgacg gtcagcaggt tgttcgttgc tactctatct ctgaccgtct gaactctgac 300
ggtctgcgtt tctctacccg tgaacgtaac gcgctggttc gtgaaatccg tggtcagaac 360
ccgaacctgc gtgaagaaga cctgaacttc gaacagtaca aagtttgcat gcacggtaaa 420
ggtaaatctc agggtgaagc gatcgcgacc gttttcgaag ttatccgtga gaaggactct 480
cagggtcgtg accgtttcgc gaaatactct gcgtctgaaa tctctctgct gcgtcacatc 540
gaacgtaacc gtctgaacgg tatcaacgcg ccggcgccgc gttctctgct gaccgttaaa 600
gaaatcggtt ctatccgtct gaaccaggac cagcgtgttc agctgggtca cctggttaac 660
ttcgttcagg ttgcgccggg tcagcagggt atcttctctt tcatggaagt tctggcgtct 720
aaccagaaaa tcaacatcga acgtggtatc aacgaaggta tcctgccgta catcacccgt 780
atctaccgtt cttacctggg ttctctgcag aacgacatcc agaaccgttc tcagaaattc 840
gaatctcacg gtttcttcct gggtctgctg gcgaacttca tccacctgta caccatcgac 900
atcgacctgg acctgtctcc gggtaactct tacgttgcgt tcctgatctg ccaccaggcg 960
gaacgtgaaa acatcccgat cgttatcaac gttacccgtt ggcgtacctc ttctgacatc 1020
gcgctgaacc gtgcgcgtgc ggacgcgaaa cgtctgcacg tttcttcttt catctctatc 1080
cacaccgaat ctcgtaacgc ggtttgcatc ggtctgaact tcaacctgaa catcgacccg 1140
ttctctatcg acaccgttga attcctggaa aaccgtttcc cgctggttca gcgtctgttc 1200
gaatgcctgg aagacgaagg tatccgtgaa aacatccgtg acttcctgct gcagcacctg 1260
ccgaacgaaa tcccgcgtaa cgcggaaaac tacaaccgta tcttcgactg catcaccggt 1320
ttcgcgttcg gtaactctat cctggaagaa ttccgtctgg ttaacgcggt tcagcagcgt 1380
gttcgtaaat acatcttccg ttacggtgac gaaaaccacg cgctgaccat ggttttccac 1440
acccagggtt ctgacatcgt tatcctgcac atccgtgaca acaacgcggt tcagcagggt 1500
gcgatcaacc tgcaggacct gaacgttgac ggtaacaacg ttcacgttcg tgaagtttct 1560
tgcaccctga acaaccagct gggtctgaac atccacaccg acaacctggg tctgtaccac 1620
aactaccaga acaacaacgc gaacaacttc ctgggtggta acctggttca ggttccgaac 1680
gcgggtaacg ttcacaacgc gctgaaccag gttatgaacg acggttggca ggaccgtttc 1740
cagcaccagg aactgttccg taacatctct gcggttctga tgccggaaga cacccacggt 1800
aacatgatca tcgacgttaa ctctaaagac aaattccgtt ctatcctgca cggtaccttc 1860
tacgcgtctg acaacccgta caaagttctg gcgatgtaca aagttggtca gacctactct 1920
ctgaaacgtt ggcaggaaga agaaggtgaa cgtgttatcc tgacccgtgt taccgaacag 1980
cgtctgggtc tgctgctgct gcgtcagccg accgcggaca cccacccgat cggttacgtt 2040
ctgggtttcg cggacaacgc ggaagaagtt gaacaggaac aggacgaagc gcgttacaaa 2100
atcaccgaac tgatgtctaa acagcgtggt tacctgccga tcacctctgg taacgaagtt 2160
gttctgtctt acgcggtttt caaccgtggt gcgcagcgtg cggaagactt catctctctg 2220
ccgcagcagg cggtttacgt tcaccgtctg gaccgtcgtg gtcacgactc tcgtccggaa 2280
gttctggttg gtccggaatc tgttatcgac gaaaacccgc cggaaaacct gctgtctgac 2340
cagacccgtg aaaacttccg tcgtttctac atggagaagc gtccgggtca gaactctatc 2400
ttcctgctgg acatcgacga caacctgcac gttccgttct cttacctgca gggtacccgt 2460
gcgcaggcga tcgaaaccct gcgttctcgt atccgtggtg gtggtacctc taccgcgcag 2520
ggtatcctgc agcagatcaa caccatcctg cgtcgtaaca acgcgcgtga aatcgaagac 2580
gttcacaacc tgctggcgct ggacttcgcg accgaaaacc agaacttccg ttactggctg 2640
cagacccacg acatgttctt cgcggcgcgt cagtacacct tccacgacga ccgttctaac 2700
ccgaccaacg accgtcacga cttcgcgatc acctctgttg gtgttgacgg taaccagaac 2760
gacccgaccg gtcgtgacct gctgtcttct aacatcgaca acttcaaaca gaaagttgac 2820
tctggtgaga aggaccgtct gaccgcgatc atcaacgttg gtaaccgtca ctgggttacc 2880
ctggttatcg ttcaccagaa cggtaactac tacggttact acgcggactc tctgggtccg 2940
gactctcgta tcgacaacaa catccgtggt gcgctgcgtg aatgcgacat ctctgacgac 3000
aacgttcacg acgtttctgt tcaccagcag accgacggtc acaactgcgg tatctgggcg 3060
tacgaaaacg cgcgtgacat caaccaggcg atcgaccagg cgctgcaggg taactctaac 3120
ttcggtgaga agggtgaagg tatcatcggt tacatccgtg gtctgctgtc tgcgggtatc 3180
ggtaacgaca cccgtcagcc gcagcgtaac gaacagtact tccgtaaccg tcgtcgtaac 3240
atctctcagc tgttccagaa cgactctctg tcttctccgc gtggtcgtct gatccagggt 3300
cgtccgggta tccagcacga aatcgacccg ctgctgctgc agttcctgga actgcagtac 3360
ccgcagcgtg gtggtggtgg tgcgctgcag ctgggtggtg aacgtgttat ctctatcgac 3420
ttcggtccgc agtctgttct ggacgaaatc gacggtgtta accgtgttta cgaccactct 3480
aacggtcgtg gttctcgt 3498
<210> 18
<211> 32
<212> DNA
<213> 人工序列()
<400> 18
ggaattccat atgtctaacg gtgacggtct ga 32
<210> 19
<211> 31
<212> DNA
<213> 人工序列()
<400> 19
ccgctcgaga cgagaaccac gagagttaga g 31
<210> 20
<211> 57
<212> DNA
<213> 人工序列()
<400> 20
cgggatccat ggactataaa gacgatgacg ataaaatgtc taacggtgac ggtctga 57
<210> 21
<211> 35
<212> DNA
<213> 人工序列()
<400> 21
acgcgtcgac ttaacgagaa ccacgagagt tagag 35
<210> 22
<211> 1335
<212> DNA
<213> 人工序列()
<400> 22
atggaatctg gtctggacca caactacaac aaaatcctgg acatcctgaa aggtgcgatc 60
aaaggtgacg acaaccaggt taaagcgcgt aaacacctgc gtgttgaacg ttggctgcgt 120
gcgtacatcc agctgatcga agacttcgac gaagagaagc tgatcttctt ctctgacatc 180
ttctctgaca actcttgctg ggacggtatc aaactgaaga ataaagcggt tggtgaacgt 240
ctgaccgaag agaagaacaa gaatggtaaa gaaaacccgc tggacctggc ggaccgttac 300
tacctggcgt gcaaatactg cctggaagac aaaatcccgg gtctgttcga acaggttttc 360
atgcgtttca aacgttctgc gttcgaagaa gacggttctg acgacgacct gcgtcgtgaa 420
ctgctggaaa acatcgaaga aacctctccg atcgaagcgt tctggtcttt cctgatcgac 480
aaacagatcg gtaaactgaa cgaatacaaa tctgttgaag gtctgcagaa atctatccag 540
atcaactcta acaagaattg ggaagaaggt atcgaattct tctacaacaa actgcacaac 600
gactcttcta tctcttctca ggacaaagac gacctgctga tcgaagcggc gctgtctgcg 660
gttaaaggtt acaaagaagt tgacaccatc gaattctgcc tgtctaaaat ggacgacgaa 720
cagaagaaga aactgctgga ccgtgactac aaagaaaaca cctactacgc ggttctgaac 780
gttctggttg gtcagtacta cttcgactct ttcatggaac tgtctcgtct gtgctctcag 840
atcgaatgcg aacgttacac caccttcctg tcttctctgt ctgaccaggt tctgaagaat 900
ccggacctgt ctgaagaaac caagaagtgc atgatgaacg tttgggaacg tatcatcaaa 960
ctgaaaaccc aggaccgtgg tgaacagtct atctcttcta tcttcgttga ctactctgtt 1020
acctacacca tcgcgaacct gatcgttgac ccgtctcgtc agggtgtttc taaagaagaa 1080
atcctgggta aaatcctgaa acacgttaaa gaaatgtctg gtgaagaaat gatcaaagtt 1140
aaagactctg ttctgtctaa aatccagctg ttccacggtg gtaagaagct gcagctgggt 1200
gaacaggttt tctctaaact ggcgcaggaa gcgtctaaag aatctatcct gcgtgaagcg 1260
ggtgacaccc tgccgcagtc ttctctgtct accaccgaca ccccgtacaa catcaaatct 1320
ctgtctcact ctaaa 1335
<210> 23
<211> 28
<212> DNA
<213> 人工序列()
<400> 23
ggaattccat atggaatctg gtctggac 28
<210> 24
<211> 31
<212> DNA
<213> 人工序列()
<400> 24
ccgctcgagt ttagagtgag acagagattt g 31
<210> 25
<211> 53
<212> DNA
<213> 人工序列()
<400> 25
cgggatccat ggactataaa gacgatgacg ataaaatgga atctggtctg gac 53
<210> 26
<211> 35
<212> DNA
<213> 人工序列()
<400> 26
acgcgtcgac ttatttagag tgagacagag atttg 35
<210> 27
<211> 2196
<212> DNA
<213> 人工序列()
<400> 27
atgccgtcta acgttaaacc gctggaactg gttcagctgc tgctgatgcg taacaaatct 60
aaagacgaat tcctggactt ccagaaacgt ttccagtctt tcatcaacca gtctccgtct 120
ttcctgcact ctgttggtaa accgggtttc ttcccgtctt tcttcttcgg tatgttcgcg 180
accgttctgg acaccgaact ggcgaccaaa atcggtatca agaagctgca cttccgtttc 240
gacgacaacc gtaccctgaa aatcgcgatc ctgaccaacg aaggtctgaa atgcatcacc 300
atgtctgacc aggttgacgg taacatgcac ctgaaattct ctcagggtga actggagaag 360
atcgcgcaga aatggaaaat gggtgcggaa ttcgacaaac tggagaagga agaacacgaa 420
atcaccatca ccggtaaaga agttaaacac ggtaaagttg acccggcgtt ctctaagaag 480
accgactact ctcagaaagg tttcaccgaa atcgagaagg accgtgacca gcaggacctg 540
gaatctctga tctctaaact gtctaaccag gacttcgaag aagttaagaa gaacgcgcgt 600
cgtatgttca actacatcac caacgtttac aagaagtacg agaaggaaac cctgttctct 660
ggtaaagaat cttctcacca cggtttcctg gcgggtttcc tgatcaactt caaataccgt 720
ttccacctga aactgtacct ggaactgttc gcgggtaaag gttacgcgga catcatcctg 780
ctggttcgtg gttctgacaa atctctgtct tctatcccga tcatcatcga actgaaagcg 840
ggtaccggtg aaatctctac cgttatcaaa gcgctgaaac aggcgcagga ctacgttaaa 900
ggttctttct ctaactctat ccgtatgatc accatcgcga acgaagcgat ctgcgttggt 960
ctgaacttcg acatggttca ccacgaaaac gttaaaatcg acgttgaaaa cttcctgtct 1020
cgtgaaggta actctgttat cgagaagctg ctgggtaccg aagcgaccaa cgcggaagtt 1080
atccgtaccc agctggaata cctgtactac ggtatcgttt ggtctaacgg tggttctgac 1140
aacatcaact acgtttctcg tatgatcctg ggtcagctgg ttctgatctc taacatcatc 1200
aaacgtgaga agctgggtaa acacatcttc atctacgacc agaacgacaa aatggttacc 1260
ggttctcaga aacgtccgga agcggcgaaa gaatctatcg aagactgcgt taccaccatc 1320
gttctgaccc tgggtaagaa ggttctgatc ctgaacatca acgagaagaa cgaattcgcg 1380
ctgcgtgttc cggacaacaa aggtatcccg atcgaaaaca tccgtcgtat ccagaacgtt 1440
aacgacatca aaatccagga aatcacctgc aacctgtact ctaccccgtc taacaagaat 1500
ccgttcgacc agtactgcaa caagaataaa ggtatcaccg ttaacaccta cgactctctg 1560
gacaaataca aacgtggtaa agaaatcctg cagggtaact tcacccgtat cgttgaaaac 1620
aagaagttca aagcggcgct gtctaaagcg atcgaatctg gtaaatacga cgactacaag 1680
aagctgttcg aagaaatctc tcacatcctg cacccgttca aatctctgat ctctaacgaa 1740
gcgaccttcc aggcggttct gcacggtctg ttctcttctt acggtgaaga caacatcaaa 1800
gttatcaccg aattccagat cggtggtggt gagaagctgg acgttatgct ggttatcaac 1860
gcgaccgacc agaagaagga atacccgccg gttggtatcg aactgaaatt cgcgaagaag 1920
ggtgaactgg acaagaagga gaaggacgcg aaagaccagc tgaaacgtta caaagaaggt 1980
gaagcgtaca aagttatcac cgacgcgggt aaagttaaac tgatctacgc ggttttcaac 2040
aaaggtgcga ccgacgaagg ttctctgatc aaaatcggta acgaattcgt tgaagttgac 2100
gttcgtcact cttctgttgt tgcgttcggt cagcagccgg gttctctgca gcagccgtac 2160
gttaaacagg cgggtctgtc tcgtgcggtt aaccag 2196
<210> 28
<211> 31
<212> DNA
<213> 人工序列()
<400> 28
ggaattccat atgccgtcta acgttaaacc g 31
<210> 29
<211> 29
<212> DNA
<213> 人工序列()
<400> 29
ccgctcgagc tggttaaccg cacgagaca 29
<210> 30
<211> 56
<212> DNA
<213> 人工序列()
<400> 30
cgggatccat ggactataaa gacgatgacg ataaaatgcc gtctaacgtt aaaccg 56
<210> 31
<211> 33
<212> DNA
<213> 人工序列()
<400> 31
acgcgtcgac ttactggtta accgcacgag aca 33
<210> 32
<211> 57
<212> DNA
<213> 人工序列()
<400> 32
cgggatccat ggactataaa gacgatgacg ataaaatgtc taacggtgac ggtctga 57
<210> 33
<211> 35
<212> DNA
<213> 人工序列()
<400> 33
acgcgtcgac ttaacgagaa ccacgagagt tagag 35
<210> 34
<211> 445
<212> PRT
<213> 人工序列()
<400> 34
Met Glu Ser Gly Leu Asp His Asn Tyr Asn Lys Ile Leu Asp Ile Leu
1 5 10 15
Lys Gly Ala Ile Lys Gly Asp Asp Asn Gln Val Lys Ala Arg Lys His
20 25 30
Leu Arg Val Glu Arg Trp Leu Arg Ala Tyr Ile Gln Leu Ile Glu Asp
35 40 45
Phe Asp Glu Glu Lys Leu Ile Phe Phe Ser Asp Ile Phe Ser Asp Asn
50 55 60
Ser Cys Trp Asp Gly Ile Lys Leu Lys Asn Lys Ala Val Gly Glu Arg
65 70 75 80
Leu Thr Glu Glu Lys Asn Lys Asn Gly Lys Glu Asn Pro Leu Asp Leu
85 90 95
Ala Arg Arg Tyr Tyr Leu Ala Cys Lys Tyr Cys Leu Glu Asp Lys Ile
100 105 110
Pro Gly Leu Phe Glu Gln Val Phe Met Arg Phe Lys Arg Ser Ala Phe
115 120 125
Glu Glu Asp Gly Ser Asp Asp Asp Leu Arg Arg Arg Leu Leu Glu Asn
130 135 140
Ile Glu Glu Thr Ser Pro Ile Arg Ala Phe Trp Ser Phe Leu Ile Asp
145 150 155 160
Lys Gln Ile Gly Lys Leu Asn Glu Tyr Lys Ser Val Glu Gly Leu Gln
165 170 175
Lys Ser Ile Gln Ile Asn Ser Asn Asp Asn Trp Glu Glu Gly Ile Glu
180 185 190
Phe Phe Tyr Asn Lys Leu His Asn Asp Ser Ser Ile Ser Ser Gln Asp
195 200 205
Lys Asp Asp Leu Leu Ile Glu Ala Ala Leu Ser Ala Val Lys Gly Tyr
210 215 220
Glu Glu Val Asp Thr Ile Glu Phe Cys Leu Ser Lys Met Asp Asp Glu
225 230 235 240
Gln Lys Lys Lys Leu Leu Asp Arg Asp Tyr Lys Glu Asn Thr Tyr Tyr
245 250 255
Ala Val Leu Asn Val Leu Val Gly Gln Tyr Tyr Phe Asp Ser Phe Met
260 265 270
Glu Leu Ser Arg Leu Cys Ser Gln Ile Glu Cys Glu Arg Tyr Thr Thr
275 280 285
Phe Leu Ser Ser Leu Ser Asp Gln Val Leu Lys Asn Pro Asp Leu Ser
290 295 300
Glu Glu Thr Lys Lys Cys Met Met Asn Val Trp Glu Arg Ile Ile Lys
305 310 315 320
Leu Lys Thr Gln Asp Arg Gly Glu Gln Ser Ile Ser Ser Ile Phe Val
325 330 335
Asp Tyr Ser Val Thr Tyr Thr Ile Ala Asn Leu Ile Val Asp Pro Ser
340 345 350
Arg Gln Gly Val Ser Lys Glu Glu Ile Leu Gly Lys Ile Leu Lys His
355 360 365
Val Lys Glu Met Ser Gly Glu Glu Met Ile Lys Val Lys Asp Ser Val
370 375 380
Leu Ser Lys Ile Gln Leu Phe His Gly Gly Lys Lys Leu Gln Leu Gly
385 390 395 400
Glu Gln Val Phe Ser Lys Leu Ala Gln Glu Ala Ser Lys Glu Ser Ile
405 410 415
Leu Arg Glu Ala Gly Asp Thr Leu Pro Gln Ser Ser Leu Ser Thr Thr
420 425 430
Asp Thr Pro Tyr Asn Ile Lys Ser Leu Ser His Ser Lys
435 440 445
<210> 35
<211> 1335
<212> DNA
<213> 人工序列()
<400> 35
atggaatctg gtctggacca caactacaac aaaatcctgg acatcctgaa aggtgcgatc 60
aaaggtgacg acaaccaggt taaagcgcgt aaacacctgc gtgttgaacg ttggctgcgt 120
gcgtacatcc agctgatcga agacttcgac gaagagaagc tgatcttctt ctctgacatc 180
ttctctgaca actcttgctg ggacggtatc aaactgaaga ataaagcggt tggtgaacgt 240
ctgaccgaag agaagaacaa gaatggtaaa gaaaacccgc tggacctggc gcgtcgttac 300
tacctggcgt gcaaatactg cctggaagac aaaatcccgg gtctgttcga acaggttttc 360
atgcgtttca aacgttctgc gttcgaagaa gacggttctg acgacgacct gcgtcgtcgt 420
ctgctggaaa acatcgaaga aacctctccg atccgtgcgt tctggtcttt cctgatcgac 480
aaacagatcg gtaaactgaa cgaatacaaa tctgttgaag gtctgcagaa atctatccag 540
atcaactcta acgataattg ggaagaaggt atcgaattct tctacaacaa actgcacaac 600
gactcttcta tctcttctca ggacaaagac gacctgctga tcgaagcggc gctgtctgcg 660
gttaaaggtt acgaagaagt tgacaccatc gaattctgcc tgtctaaaat ggacgacgaa 720
cagaagaaga aactgctgga ccgtgactac aaagaaaaca cctactacgc ggttctgaac 780
gttctggttg gtcagtacta cttcgactct ttcatggaac tgtctcgtct gtgctctcag 840
atcgaatgcg aacgttacac caccttcctg tcttctctgt ctgaccaggt tctgaagaat 900
ccggacctgt ctgaagaaac caagaagtgc atgatgaacg tttgggaacg tatcatcaaa 960
ctgaaaaccc aggaccgtgg tgaacagtct atctcttcta tcttcgttga ctactctgtt 1020
acctacacca tcgcgaacct gatcgttgac ccgtctcgtc agggtgtttc taaagaagaa 1080
atcctgggta aaatcctgaa acacgttaaa gaaatgtctg gtgaagaaat gatcaaagtt 1140
aaagactctg ttctgtctaa aatccagctg ttccacggtg gtaagaagct gcagctgggt 1200
gaacaggttt tctctaaact ggcgcaggaa gcgtctaaag aatctatcct gcgtgaagcg 1260
ggtgacaccc tgccgcagtc ttctctgtct accaccgaca ccccgtacaa catcaaatct 1320
ctgtctcact ctaaa 1335

Claims (3)

1.CifA和CifB突变蛋白用于调控胞质不相容性的用途。
2.根据权利要求1的用途，所述CifA和CifB突变蛋白来源于沃尔巴克氏体，其特征在于所述突变蛋白相对于亲本存在若干个氨基酸的突变。
3.根据权利要求1的用途，其特征是所述突变蛋白包括但不限于列举的几种，其中CidAST的氨基酸序列是SEQ ID NO.2所示，CidBST的氨基酸序列是SEQ ID NO.4所示，CinAST的氨基酸序列是SEQ ID NO.6所示，CinBST的氨基酸序列是SEQ ID NO.8所示。
CN202111665809.2A 2021-12-31 2021-12-31 CifA和CifB突变蛋白用于调控胞质不相容性的用途 Pending CN114395018A (zh)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017214476A1 (en) * 2016-06-09 2017-12-14 Vanderbilt University Cytoplasmic incompatibility factors and methods for controlling arthropods
CN108018309A (zh) * 2012-03-05 2018-05-11 奥西泰克有限公司 生物防治

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108018309A (zh) * 2012-03-05 2018-05-11 奥西泰克有限公司 生物防治
WO2017214476A1 (en) * 2016-06-09 2017-12-14 Vanderbilt University Cytoplasmic incompatibility factors and methods for controlling arthropods

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JASON L. RASGON等: "Evolution-guided mutagenesis of the cytoplasmic incompatibility proteins: Identifying CifA’s complex functional repertoire and new essential regions in CifB", PLOS PATHOG, vol. 16, no. 8, pages 1 - 18 *
YUNJIE XIAO 等: "Structural and mechanistic insights into the complexes formed by Wolbachia cytoplasmic incompatibility factors", PNAS, vol. 118, no. 41, pages 1 - 10 *

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