CN108026149A - 工程化的cry6a杀虫蛋白 - Google Patents

Abstract

公开了工程化的和修饰的Cry6Aa杀虫毒素，编码这些毒素的多核苷酸，这些毒素控制害虫的用途，以及产生这些毒素的转基因植物。

Description

工程化的CRY6A杀虫蛋白

技术领域

本发明整体涉及应用于农业科学的分子生物学领域。更具体地，某些实施方案涉及使用DNA区段作为杀虫蛋白表达的诊断探针和模板的方法。公开了制备和在转基因植物细胞和植物中结合保护剂的植物研发中使用所要求保护的核酸区段的方法。

背景技术

昆虫和其他害虫使农户每年在作物损失和控制害虫的花费方面耗损数十亿美元。在农业生产环境中由害虫引起的损失包括作物产量降低、作物品质降低，以及收获成本增加。

鞘翅目昆虫是一群大量的农业害虫，其每年造成对作物的广泛损害。鞘翅目害虫的实例包括但不限于：科罗拉多马铃薯甲虫(CPB)、玉米根虫，苜蓿叶象甲、棉铃象甲，以及日本丽金龟。科罗拉多马铃薯甲虫是一种经济上重大的害虫，其以马铃薯、茄子、番茄、辣椒、烟草和茄科的其他植物的叶子为食。科罗拉多马铃薯甲虫是马铃薯的难对付的食叶虫，这部分是因为它已经发展出了对许多类别的杀虫剂的抗性。

西方玉米根虫(WCR)(玉米根萤叶甲(Diabrotica virgifera virgiferaLeConte))是北美最具破坏性的鞘翅目昆虫之一，在美国中西部玉米种植区备受关注。每年有约930万英亩的美国玉米被玉米根虫物种复合体感染。北方玉米根虫(NCR)(巴氏根萤叶甲(Diabrotica barberi Smith and Lawrence))是与WCR在几乎相同的范围内共栖的近缘物种。还有另外若干种叶甲属(Diabrotica)的相关亚种是美洲的严重害虫：墨西哥玉米根虫(MCR)(墨西哥玉米根萤叶甲(D.virgifera zeae Krysan and Smith))；南方玉米根虫(SCR)(十一星根萤叶甲(D.undecimpunctata howardi Barber))；黄瓜条根萤叶甲(D.balteata LeConte)；黄瓜十一星叶甲(D.undecimpunctata tenella)；南美叶甲(D.speciosa Germar)；以及D.u.undecimpunctata Mannerheim。美国农业部已估计玉米根虫每年造成10亿美元收益损失，包括8亿美元的产量损失和2亿美元的治理成本。

WCR卵和NCR卵均在夏季期间沉积于土壤中。这些昆虫在整个冬季都停留在卵期。幼虫在五月底或六月初孵化并开始以玉米根为食。玉米根虫历经三个幼虫龄期。进食几周后，幼虫蜕皮，进入蛹期。它们在土壤中化蛹，然后在七月和八月以成虫形式从土壤中出现。

玉米中大部分的根虫损害由幼虫进食引起。新孵出的根虫最初以纤细的玉米根毛为食，并钻入根尖内。随着幼虫长得更大，它们以初生根为食并钻入其中。在有大量玉米根虫时，幼虫进食时常导致根修剪，一直到玉米杆的基部。严重的根损伤妨碍根将水和养分运输到植物中的能力、减缓植物生长，并导致籽粒产生减少，从而时常大幅降低总产量。严重的根损伤还时常导致玉米植物倒伏，这使收获变得更加困难，并进一步降低产量。此外，成虫以玉米生殖组织为食可导致穗尖处的穗丝修剪。如果这种“穗丝剪切”在花粉脱落期间足够严重，则传粉可能受到破坏。此外，叶甲(Diabrotica)属的成员攻击瓜类作物(黄瓜、甜瓜、南瓜等)和商业生产中的许多蔬菜和大田作物以及在家庭花园中生长的那些作物。

已经通过作物轮作、化学杀虫剂、生物杀虫剂(例如，形成孢子的革兰氏阳性细菌苏云金芽孢杆菌(Bacillus thuringiensis)(B.t.)、表达B.t.毒素的转基因植物或这些手段的组合，来尝试控制玉米根虫。作物轮作的缺陷是不必要地限制了农田的用途。另外，一些根虫物种可能在大豆田间产卵，从而损害玉米和大豆实施的作物轮作的效率。

化学杀虫剂是最为人们所倚重的用于实现玉米根虫控制的策略。使用化学杀虫剂并不是完美的玉米根虫控制策略；大的幼虫种群、暴雨以及杀虫剂应用不当都可导致玉米根虫控制不充分。此外，连续使用杀虫剂可能选择杀虫剂抗性根虫品种，并且由于杀虫剂对非靶标物种存在毒性，所以有严重的影响环境之虞。

由线虫动物造成的植物损害也是一个普遍和严重的经济问题。线虫动物引起许多植物物种中的广泛和严重损害。已知许多线虫属引起这种损伤。植物寄生的线虫动物包括线虫动物门，垫刃目和矛线目(Dorylaimide)的成员。在垫刃目中，植物寄生的线虫动物存在于以下两个超科中：垫刃总科和环总科。存在超过100,000种被描述的线虫动物物种。

化学杀虫剂已经提供了一种有效的害虫控制方法；然而，公众已经变得关注在食物、地下水和环境中可能存在的残留化学品的量。对化学杀虫剂使用的严格新限制和从市场上消除一些有效杀虫剂可能限制用于控制造成损失大的害虫的经济和有效的选择。因此，迫切需要鉴定新的害虫控制剂和组合物。

经常使用化学杀虫剂来控制不希望的害虫可能选择化学抗性品种。化学抗性发生在许多经济上重要的昆虫物种中，并且也发生在绵羊、山羊和马的线虫动物中。例如，用于控制线虫动物的公认方法集中围绕药物苯并咪唑及其同源物。这些药物的大规模使用已经导致了线虫动物种群中的许多抗性实例(Prichard，R.K.等人)。杀虫剂抗性的发展使得需要继续寻找具有不同作用模式的新型控制剂。

目前，需要有更有效的手段来控制对易感的宿主和作物造成相当大的损害的许多鞘翅目昆虫和线虫动物。有利地，这种有效手段将采用高度选择性的生物毒素。若干种B.t.Cry蛋白已表现为杀线虫动物的，这些蛋白包括Cry5B、Cry6A、Cry14A和Cry21A(Wei等人，2003；Aroian和Li(2010))。

B.t.是一种土壤传播的细菌，其产生被称为δ内毒素或Cry蛋白的杀虫晶体蛋白。Cry蛋白是通过作用于易感昆虫的中肠细胞起作用的口服毒物。一些Cry毒素已显示为具有抗线虫动物的活性。δ内毒素的广泛列表在由Neil Crickmore维护的苏云金芽孢杆菌(Bacillus thuringiensis)毒素命名学网站上维持和定期更新。(参见http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/toxins2.html和Crickmore等人，1998，第808页)。Cry毒素，包括Cry1B、Cry1I、Cry2A、Cry3、Cry7A、Cry8、Cry9D、Cry14、Cry18、Cry22、Cry23、Cry34、Cry35、Cry36、Cry37、Cry43、Cry55、Cyt1A和Cyt2C(Frankenhuyzen，2009)等家族的成员，具有针对鞘翅目昆虫的杀虫活性。

具有抗玉米根虫和其他鞘翅目昆虫活性的一些B.t.毒素是现在已知的。已经报道Cry6Aa具有抗鞘翅目昆虫和线虫害虫的活性(美国专利No.5,186,934；美国专利No.6,632,792B2；美国专利No.2011/0225681；美国专利No.2011/0239334A1；和Wei等人，2003)。例如，美国专利No.4,849,217公开了各种分离物，包括PS52A1和PS86A1，具有抗苜蓿叶象甲的活性。美国专利No.5,208,017公开PS86A1具有抗西方玉米根虫的活性。美国专利No.5,427,786和No.5,186,934各自公开了具有抗鞘翅目昆虫活性的B.t.分离物和毒素。在这些专利中特别公开的是称为PS86A1的分离物和可从其获得的称为86A1的具有抗鞘翅目昆虫活性的毒素。毒素86A1现在也被称为Cry6A(CryVIA)。野生型Cry6Aa毒素为约54.1kDa。Cry6B毒素也是已知的。这种毒素可以从PS69D1分离物获得。Cry6Aa被认知为针对西方玉米根虫的新型作用模式，补充Cry3Aa和Cry34Ab1/Cry35Ab1(Li等人，2013)使其成为综合昆虫抗性管理程序中的金字塔配偶体(美国专利No.2013/0167269A1和US 2013/0263331A1)。

已知全长Cry6A和Cry6B毒素具有抗线虫动物的活性。已报道PS69D1分离物具有抗线虫动物的活性(美国专利No.4,948,734；No.5,093,120；No.5,262,399；以及No.5,439,881)。在WO 92/19739中已经公开了CryVI毒素的氨基酸序列的通式，该专利还教导了全长毒素具有抗线虫动物的活性。其中公开了PS52A1和PS69D1分离物。美国专利No.5,262,159和No.5,468,636也公开了具有抗蚜虫活性的毒素的通式。

已知Cry6A毒素抑制某些鞘翅目昆虫的生长，并且可通过酶催化分裂而活化以产生对鞘翅目昆虫如西方玉米根虫致死的氨基末端核心毒素(美国专利No.6,831,062B2)。此外，截短的Cry6A具有抗线虫动物的活性。美国专利No.6,831,062描述了Cry6A截短的全毒素和融合蛋白以及融合基因。Thompson等人公开了根据切割位点鉴定为残基12-390和12-443的具有杀虫活性的肽片段。由胰蛋白酶或其他蛋白水解消化产生的约残基12-390或12-443的大片段被称为核心片段或毒素。对由重组B.t.产生的Cry6Aa进行胰蛋白酶处理提高了抗WCR的活性(美国专利No.5,874,288；美国专利No.6,831,062B2；以及美国专利No.6,303,364B1)。

尽管在转基因植物中产生目前部署的Cry蛋白可以提供针对上述害虫的稳健保护，从而保护谷物产量，但是害虫成虫已经出现在人工感染试验中，表明不完全的昆虫幼虫控制。此外，抗性昆虫种群的发展威胁Cry蛋白在害虫防治中的长期耐久性。鞘翅目昆虫已经在田间发展出了对Cry蛋白的抗性(Gassman等人，PLoS ONE，2011年7月|第6卷|第7期|e22629)。昆虫对B.t.Cry蛋白的抗性可以通过若干种机制发展(Heckel等人，2007；Pigott和Ellar，2007)。已经在昆虫内鉴定出Cry蛋白的多种受体蛋白类别，并且在每种受体类别中存在多个实例。例如，借助于受体蛋白的钙粘蛋白结构域的毒素结合部分内的突变，可以发展出对特定Cry蛋白的抗性。另一种抗性手段可以通过原毒素加工蛋白酶介导。

尽管在B.t.品种中自然表达的天然的Cry6Aa已显示出抗WCR和某些线虫动物的良好效力，但其作为结合到植物中的有效保护剂的用途由于其在植物细胞中表达时易受蛋白水解的影响而尚未得到证实。因此，非常需要将当在植物细胞中表达时更抗蛋白水解的工程化的Cry6A毒素用于重组植物，特别是玉米中，以作为结合到植物中的保护剂。

发明内容

本发明包括工程化的基于Cry6A的杀虫蛋白毒素，包括部分设计用于限制重要羧基末端肽(CTP)损失的变体和类似物。本发明的其他实施方案包括编码所要求保护的杀虫毒素的核酸，使用从该要求保护的核酸序列表达的毒素来控制害虫的方法，在转基因宿主细胞中产生毒素的方法，以及包含此类核酸的转基因植物种子，以及表达所述毒素的植物。

本发明的经修饰的Cry6Aa杀虫蛋白包含选自由以下项组成的组的修饰：经修饰的蛋白水解易感区域(SEQ ID NO:1的残基390-451)，CTP对核心蛋白的亲和力增加，以及亚细胞转运肽的添加。优选的一组Cry6Aa变体杀虫蛋白是SEQ ID NO:35、SEQ ID NO:37、SEQ IDNO:39、SEQ ID NO:41、SEQ ID NO:43、SEQ ID NO:45、SEQ ID NO:47、SEQ ID NO:49、SEQ IDNO:64、SEQ ID NO:66、SEQ ID NO:68、SEQ ID NO:70、SEQ ID NO:72、SEQ ID NO:74、SEQ IDNO:76、SEQ ID NO:78、SEQ ID NO:80、SEQ ID NO:82、SEQ ID NO:84、SEQ ID NO:86、SEQ IDNO:88、SEQ ID NO:90、SEQ ID NO:92、SEQ ID NO:94、SEQ ID NO:96、SEQ ID NO:98、SEQ IDNO:100、SEQ ID NO:102、SEQ ID NO:104、SEQ ID NO:106、SEQ ID NO:108、SEQ ID NO:110、SEQ ID NO:112、SEQ ID NO:114、SEQ ID NO:116、SEQ ID NO:118、SEQ ID NO:120、SEQ IDNO:132、SEQ ID NO:136、SEQ ID NO:138和SEQ ID NO:140。更优选的一组变体是SEQ IDNO:110、SEQ ID NO:112、SEQ ID NO:114、SEQ ID NO:116、SEQ ID NO:118、SEQ ID NO:120、SEQ ID NO:128、SEQ ID NO:132和SEQ ID NO:144。更加高度优选的一组变体是SEQ ID NO:110、SEQ ID NO:116、SEQ ID NO:118、SEQ ID NO:120、SEQ ID NO:132。甚至更优选的一组变体是SEQ ID NO:116、SEQ ID NO:118、SEQ ID NO:120。并且最优选的变体是SEQ ID NO:116。

编码Cry6Aa变体杀虫蛋白的优选的一组核酸序列是SEQ ID NO:34、SEQ ID NO:36、SEQ ID NO:38、SEQ ID NO:40、SEQ ID NO:42、SEQ ID NO:44、SEQ ID NO:46、SEQ IDNO:48、SEQ ID NO:63、SEQ ID NO:65、SEQ ID NO:67、SEQ ID NO:69、SEQ ID NO:71、SEQ IDNO:73、SEQ ID NO:75、SEQ ID NO:77、SEQ ID NO:79、SEQ ID NO:81、SEQ ID NO:83、SEQ IDNO:85、SEQ ID NO:87、SEQ ID NO:89、SEQ ID NO:91、SEQ ID NO:93、SEQ ID NO:95、SEQ IDNO:97、SEQ ID NO:99、SEQ ID NO:101、SEQ ID NO:103、SEQ ID NO:105、SEQ ID NO:107、SEQ ID NO:109、SEQ ID NO:111、SEQ ID NO:113、SEQ ID NO:115、SEQ ID NO:117、SEQ IDNO:119、SEQ ID NO:131、SEQ ID NO:135、SEQ ID NO:137、SEQ ID NO:139、SEQ ID NO:141、SEQ ID NO:142和SEQ ID NO:143。更优选的一组核酸是SEQ ID NO:109、SEQ ID NO:111、SEQ ID NO:113、SEQ ID NO:115、SEQ ID NO:117、SEQ ID NO:119、SEQ ID NO:127和SEQ IDNO:131。更加高度优选的一组核酸是SEQ ID NO:109、SEQ ID NO:115、SEQ ID NO:117、SEQID NO:119、SEQ ID NO:131和SEQ ID NO:139。甚至更优选的一组核酸是SEQ ID NO:115、SEQ ID NO:117、SEQ ID NO:119和SEQ ID NO:139。并且最优选的核酸是SEQ ID NO:115。

在另一个实施方案中，本发明提供了一种包含本文公开的工程化的Cry6Aa杀虫多肽的植物。

在另一个实施方案中，本发明提供了一种通过在植物组织中递送本发明的杀虫有效量的杀虫蛋白使得害虫摄入杀虫蛋白，来控制植物害虫种群和对植物的损害的方法。

在另一个实施方案中，本发明提供了一种编码本文公开的工程化的Cry6Aa杀虫多肽的分离核酸。

在另一个实施方案中，本发明提供了包含编码工程化的Cry6Aa杀虫多肽的核苷酸序列的DNA构建体，该核苷酸序列可操作地连接到能够驱动在植物中表达的启动子和稳定化信使RNA的其他调节序列。启动子可以是对于B.t.异源或天然的。

本发明还提供了包含稳定结合到植物基因组中的DNA构建体的转基因植物和一种用于保护植物免受害虫伤害的方法，该方法包括将构建体引入所述植物中。

附图说明

图1是如实施例3中所述测试的DIG-177(Cry6Aa)样品1-4的示意图。该图示出了胰蛋白酶消化后具有和不具有二硫键还原的样品。

图2是代表SEQ ID NO:2的残基12-125、128-387和445-472的经胰蛋白酶处理的DIG-177(Cry6Aa)(左)的带状3D结构模型。图2(右)是SEQ ID NO:2的残基12-472的带状模型，从胰蛋白酶处理的结构(左)中缺失的残基已经被模型化。这两种结构中的羧基末端肽均以黑色示出。

图3是对残基12-472(左侧；如来自图2)和变体DIG-1000(右侧)的结构模型比较的描述。羧基末端肽(CTP)在左侧以黑色示出；CTP和工程化的接头在DIG-1000中以黑色示出(右侧)。

图4示出对在实施例10中描述的条件下用蛋白酶K处理过的DIG-177和DIG-1000的SDS-PAGE分析。FL指代未处理、未消化的全长蛋白质；0-50是以分钟计的孵育时间。

图5是Cry6Aa蛋白的蛋白质印记玉蜀黍瞬时表达分析。所用的抗血清对DIG-177、Cry6Aa是特异性的。注意到由轰击的质粒DNA编码的蛋白质为：泳道1，荧光彩虹标记物(GE医疗集团(GE Healthcare)，宾夕法尼亚州匹兹堡)；泳道2，非轰击的胚胎对照；泳道3，具有TraP8的DIG-177；泳道4，DIG-177；泳道5DIG-1000；泳道6，具有TraP8的DIG-1000；泳道7，PAT(阴性对照)；泳道8，YFP(阴性对照)；泳道9，0.5ng/泳道的DIG-177标准品；泳道10，1ng/泳道的DIG-1000标准品。

图6示出了T₁玉蜀黍叶表达的蛋白质印记实验的结果。

序列简述

SEQ ID NO:1是包含全长DIG-177(Cry6Aa)的DNA序列。

SEQ ID NO:2是推导的DIG-177蛋白序列。

SEQ ID NO:3是DIG-177的玉蜀黍密码子优化版本，也称为IRDIG.522.28；编码鉴定为SEQ ID NO:2的蛋白质。

SEQ ID NO:4是具有编码的线粒体转运肽的DIG-177的玉蜀黍密码子偏倚版本，也称为IRDIG.552.60。

SEQ ID NO:5是由SEQ ID NO:4编码的多肽

SEQ ID NO:6是有编码的线粒体转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.61

SEQ ID NO:7是由SEQ ID NO:6编码的多肽

SEQ ID NO:8是具有编码的组合线粒体和叶绿体转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.62

SEQ ID NO:9是由SEQ ID NO:8编码的多肽

SEQ ID NO:10是具有编码的过氧化物酶转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.63

SEQ ID NO:11是由SEQ ID NO:10编码的多肽

SEQ ID NO:12是具有编码的过氧化物酶转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.64

SEQ ID NO:13是由SEQ ID NO:12编码的多肽

SEQ ID NO:14是具有编码的过氧化物酶转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.65

SEQ ID NO:15是由SEQ ID NO:14编码的多肽

SEQ ID NO:16是具有编码的液泡转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.66

SEQ ID NO:17是由SEQ ID NO:16编码的多肽

SEQ ID NO:18是具有编码的液泡转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.67

SEQ ID NO:19是由SEQ ID NO:18编码的多肽

SEQ ID NO:20是具有编码的液泡转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.68

SEQ ID NO:21是由SEQ ID NO:20编码的多肽

SEQ ID NO:22是具有编码的质外体转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.69

SEQ ID NO:23是由SEQ ID NO:22编码的多肽

SEQ ID NO:24是具有编码的内质网转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.70

SEQ ID NO:25是由SEQ ID NO:24编码的多肽

SEQ ID NO:26是具有编码的细胞核转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.71

SEQ ID NO:27是由SEQ ID NO:26编码的多肽

SEQ ID NO:28是具有编码的细胞核转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.72

SEQ ID NO:29是由SEQ ID NO:28编码的多肽

SEQ ID NO:30是具有编码的叶绿体转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.73

SEQ ID NO:31是由SEQ ID NO:30编码的多肽

SEQ ID NO:32是具有编码的叶绿体转运肽的DIG-177的玉蜀黍密码子优化版本；也称为IRDIG.552.74

SEQ ID NO:33是由SEQ ID NO:32编码的多肽

SEQ ID NO:34是DIG-177编码序列，编码突变C88>A，也称为DIG-614

SEQ ID NO:35是由SEQ ID NO:34编码的多肽

SEQ ID NO:36是DIG-177编码序列，编码突变C88>S，也称为DIG-615

SEQ ID NO:37是由SEQ ID NO:36编码的多肽

SEQ ID NO:38是DIG-177编码序列，编码突变C162>A，也称为DIG-616

SEQ ID NO:39是由SEQ ID NO:38编码的多肽

SEQ ID NO:40是DIG-177编码序列，编码突变C162>S，也称为DIG-617

SEQ ID NO:41是由SEQ ID NO:40编码的多肽

SEQ ID NO:42是DIG-177编码序列，编码突变C451>A，也称为DIG-618

SEQ ID NO:43是由SEQ ID NO:42编码的多肽

SEQ ID NO:44是DIG-177编码序列，编码突变C451>S，也称为DIG-619

SEQ ID NO:45是由SEQ ID NO:44编码的多肽

SEQ ID NO:46是DIG-177编码序列，编码突变C88>S和C451>S，也称为DIG-983

SEQ ID NO:47是由SEQ ID NO:46编码的多肽

SEQ ID NO:48是DIG-177编码序列，编码突变C88>A和C451>A，也称为DIG-984

SEQ ID NO:49是由SEQ ID NO:48编码的多肽

SEQ ID NO:50是LF(大片段)；SEQ ID NO:2的氨基酸12-390

SEQ ID NO:51是CTP 1(羧基末端肽1)，SEQ ID NO:2的氨基酸444-475

SEQ ID NO:52是CTP 2(羧基末端肽2)，SEQ ID NO:2的氨基酸451-475

SEQ ID NO:53编码由SEQ ID NO:2的残基1-443构成的蛋白质，也称为DIG-137

SEQ ID NO:54是由SEQ ID NO:53编码的多肽

SEQ ID NO:55编码由SEQ ID NO:2的残基1-432构成的蛋白质，也称为DIG-138

SEQ ID NO:56是由SEQ ID NO:55编码的多肽

SEQ ID NO:57编码由SEQ ID NO:2的残基1-423构成的蛋白质，也称为DIG-147

SEQ ID NO:58是由SEQ ID NO:57编码的多肽

SEQ ID NO:59编码由SEQ ID NO:2的残基1-400构成的蛋白质，也称为DIG-148

SEQ ID NO:60是由SEQ ID NO:59编码的多肽

SEQ ID NO:61编码由SEQ ID NO:2的残基1-390构成的蛋白质，也称为DIG-149

SEQ ID NO:62是由SEQ ID NO:61编码的多肽

SEQ ID NO:63编码缺失SEQ ID NO:2的氨基酸残基391-395的蛋白质，称为DIG-921

SEQ ID NO:64是由SEQ ID NO:63编码的多肽

SEQ ID NO:65编码缺失SEQ ID NO:2的氨基酸残基396-400的蛋白质，称为DIG-922

SEQ ID NO:66是由SEQ ID NO:65编码的多肽

SEQ ID NO:67编码缺失SEQ ID NO:2的氨基酸残基401-405的蛋白质，称为DIG-923

SEQ ID NO:68是由SEQ ID NO:67编码的多肽

SEQ ID NO:69编码缺失SEQ ID NO:2的氨基酸残基406-410的蛋白质，称为DIG-924

SEQ ID NO:70是由SEQ ID NO:69编码的多肽

SEQ ID NO:71编码缺失SEQ ID NO:2的氨基酸残基406-410的蛋白质，称为DIG-925

SEQ ID NO:72是由SEQ ID NO:71编码的多肽

SEQ ID NO:73编码缺失SEQ ID NO:2的氨基酸残基416-420的蛋白质，称为DIG-926

SEQ ID NO:74是由SEQ ID NO:73编码的多肽

SEQ ID NO:75编码缺失SEQ ID NO:2的氨基酸残基421-425的蛋白质，称为DIG-927

SEQ ID NO:76是由SEQ ID NO:75编码的多肽

SEQ ID NO:77编码缺失SEQ ID NO:2的氨基酸残基441-445的蛋白质，称为DIG-931

SEQ ID NO:78是由SEQ ID NO:77编码的多肽

SEQ ID NO:79编码缺失SEQ ID NO:2的氨基酸残基391-400的蛋白质，称为DIG-969

SEQ ID NO:80是由SEQ ID NO:79编码的多肽

SEQ ID NO:81编码缺失SEQ ID NO:2的氨基酸残基401-410的蛋白质，称为DIG-970

SEQ ID NO:82是由SEQ ID NO:81编码的多肽

SEQ ID NO:83编码缺失SEQ ID NO:2的氨基酸残基411-420的蛋白质，称为DIG-971

SEQ ID NO:84是由SEQ ID NO:83编码的多肽

SEQ ID NO:85编码缺失SEQ ID NO:2的氨基酸残基421-430的蛋白质，称为DIG-972

SEQ ID NO:86是由SEQ ID NO:85编码的多肽

SEQ ID NO:87编码缺失SEQ ID NO:2的氨基酸残基431-440的蛋白质，称为DIG-973

SEQ ID NO:88是由SEQ ID NO:87编码的多肽

SEQ ID NO:89编码缺失SEQ ID NO:2的氨基酸残基391-405的蛋白质，称为DIG-985

SEQ ID NO:90是由SEQ ID NO:89编码的多肽

SEQ ID NO:91编码缺失SEQ ID NO:2的氨基酸残基406-420的蛋白质，称为DIG-986

SEQ ID NO:92是由SEQ ID NO:91编码的多肽

SEQ ID NO:93编码缺失SEQ ID NO:2的氨基酸残基421-435的蛋白质，称为DIG-987

SEQ ID NO:94是由SEQ ID NO:93编码的多肽

SEQ ID NO:95编码缺失SEQ ID NO:2的氨基酸残基429-443的蛋白质，称为DIG-988

SEQ ID NO:96是由SEQ ID NO:95编码的多肽

SEQ ID NO:97编码缺失SEQ ID NO:2的氨基酸残基391-410的蛋白质，称为DIG-989

SEQ ID NO:98是由SEQ ID NO:97编码的多肽

SEQ ID NO:99编码缺失SEQ ID NO:2的氨基酸残基411-430的蛋白质，称为DIG-990

SEQ ID NO:100是由SEQ ID NO:99编码的多肽

SEQ ID NO:101编码缺失SEQ ID NO:2的氨基酸残基424-443的蛋白质，称为DIG-991

SEQ ID NO:102是由SEQ ID NO:101编码的多肽

SEQ ID NO:103编码缺失SEQ ID NO:2的氨基酸残基391-415的蛋白质，称为DIG-992

SEQ ID NO:104是由SEQ ID NO:103编码的多肽

SEQ ID NO:105编码缺失SEQ ID NO:2的氨基酸残基415-440的蛋白质，称为DIG-993

SEQ ID NO:106是由SEQ ID NO:105编码的多肽

SEQ ID NO:107编码缺失SEQ ID NO:2的氨基酸残基419-443的蛋白质，称为DIG-994

SEQ ID NO:108是由SEQ ID NO:107编码的多肽

SEQ ID NO:109编码缺失SEQ ID NO:2的氨基酸残基401-443的蛋白质，称为DIG-995

SEQ ID NO:110是由SEQ ID NO:109编码的多肽

SEQ ID NO:111编码缺失SEQ ID NO:2的氨基酸残基391-433的蛋白质，称为DIG-996

SEQ ID NO:112是由SEQ ID NO:111编码的多肽

SEQ ID NO:113编码缺失SEQ ID NO:2的氨基酸残基391-414和425-443的蛋白质，称为DIG-997

SEQ ID NO:114是由SEQ ID NO:113编码的多肽

SEQ ID NO:115编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1000。

SEQ ID NO:116是由SEQ ID NO:115编码的多肽

SEQ ID NO:117编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1049

SEQ ID NO:118是由SEQ ID NO:117编码的多肽

SEQ ID NO:119编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1052

SEQ ID NO:120是由SEQ ID NO:119编码的多肽

SEQ ID NO:121编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1038

SEQ ID NO:122是由SEQ ID NO:121编码的多肽

SEQ ID NO:123编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1055

SEQ ID NO:124是由SEQ ID NO:123编码的多肽

SEQ ID NO:125编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1039

SEQ ID NO:126是由SEQ ID NO:125编码的多肽

SEQ ID NO:127编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1056

SEQ ID NO:128是由SEQ ID NO:127编码的多肽

SEQ ID NO:129编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1040

SEQ ID NO:130是由SEQ ID NO:129编码的多肽

SEQ ID NO:131编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1057

SEQ ID NO:132是由SEQ ID NO:131编码的多肽

SEQ ID NO:133编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1041

SEQ ID NO:134是由SEQ ID NO:133编码的多肽

SEQ ID NO:135编码具有蛋白酶易感区段的肽接头替代的DIG-177变体，也称为DIG-1058

SEQ ID NO:136是由SEQ ID NO:135编码的多肽

SEQ ID NO:137是DIG-1000的玉蜀黍密码子偏倚版本。

SEQ ID NO:138是由SEQ ID NO:137编码的多肽

SEQ ID NO:139是具有叶绿体转运肽的DIG-1000的玉蜀黍密码子偏倚版本

SEQ ID NO:140是由SEQ ID NO:139编码的多肽

SEQ ID NO:141是DIG-1036的玉蜀黍密码子偏倚版本。

SEQ ID NO:142是具有ER定位序列的DIG-1036的最高GC+ERLS玉蜀黍最高GC版本

SEQ ID NO:143是具有液泡定位序列的DIG-1036的最高GC+VLS玉蜀黍最高GC版本

SEQ ID NO:144是由DIG-1036编码的多肽(DIG-1000-糖基化位点：N69>Q；N144>Q；N403>Q；N409>Q)

具体实施方式

本发明是设计用于探索对植物细胞中产生的Cry6A毒素的蛋白水解损伤的来源和靶位点的蛋白质工程研究的结果。已经完成了高分辨率分析，包括Cry6Aa胰蛋白酶消化的晶体结构的测定。这项工作公开了关于Cry6Aa杀虫蛋白如何发挥功能的若干个令人惊讶的分子细节。

在这些发现中，基于结构同一性而确定Cry6Aa属于蛋白质的α螺旋溶血素家族；该组的其他成员包括HlyE和BLB-B(Eifler等人，2006；Tzokov等人，2006；Wallace等人，2000；Mueller等人，2009；Madegowda等人，2008)。

鉴定了在半胱氨酸残基88与451之间以及在半胱氨酸402-404之间的两个二硫键。最令人惊讶的是，这些二硫键不是全长Cry6Aa在基于人工饮食的WCR生物测定中的杀虫活性所必需的。

第三个发现与胰蛋白酶处理的毒素的杀虫活性有关。发现由残基444-475或451-475组成的羧基末端肽(CTP)被二硫键键合到核心毒素片段，并且被发现是活性所需的。CTP的小尺寸和二硫键的不稳定性质可能导致其先前未被鉴定(美国专利No.5,874,288；美国专利No.6,831,062；美国专利No.6,303,364)。

Cry6Aa作为植物中的稳健转基因性状(结合到植物中的保护剂)的实用性，特别是用于抗WCR的保护的实用性，取决于活性杀虫蛋白的细胞内积累。在还原环境如植物细胞中残基390-451之间的蛋白水解切割据信引起羧基末端肽的解离，从而导致杀虫活性的丧失。

防止植物表达的Cry6Aa毒素的杀虫活性丧失的若干种手段形成了本发明的基础。这些手段包含减少主题毒素在易感区域(例如在大致残基390-451之间的区域)中的蛋白水解，或最小化必需的CTP的溶解，并且可以单独或组合地使用。这些手段如下：

1.重新设计易感区域的一级氨基酸结构，使其不被蛋白酶识别，包括缩短易感区域使其不易受蛋白酶影响。

2.将表达的Cry6Aa毒素重定向到蛋白酶不能进入的亚细胞或细胞外隔室。

3.下调或抑制主题毒素积累的细胞环境中的蛋白酶。

4.通过将表达的Cry6Aa重定向到内质网来允许核心-CTP二硫键保持完整，以维持CTP与核心蛋白的缔合。在这些情况下，Cry6Aa蛋白在蛋白水解后保留杀虫活性。

5.在蛋白水解发生的情况下，通过蛋白质工程化来增加核心蛋白与CTP之间的亲和力。这些方法在蛋白质工程化领域中是众所周知的，并且包括核心与CTP多肽之间的工程化氢键。另一种方式是工程化在非共价连接的肽之间的盐桥。第三种手段是工程化在CTP与核心蛋白之间的疏水相互作用。第四种方法是工程化新的或现有的二硫键，使它们不易被还原。

通过“分离的”，申请人意谓核苷酸或多肽分子已经从它们的天然环境中取出，并且已经通过人手放置在不同的环境中。因此，分离的核苷酸和多肽分子包括已经纯化、浓缩或以其他方式使得基本上不含苏云金芽孢杆菌(Bacillus thuringiensis)细胞材料的DNA或蛋白质分子。分离的工程化的Cry6Aa杀虫多肽或核苷酸分子的实施方案可以具有小于约30％、小于约20％、小于约10％、小于约9％、小于约8％、小于约7％、小于约6％、小于约5％、小于约4％、小于约3％或小于约2％、或小于约1％(按干重计)的污染蛋白(例如，来自苏云金芽孢杆菌)当分离的工程化的Cry6Aa杀虫多肽或核苷酸实施方案是重组产生的时，则培养基材料、化学前体和/或非工程化的Cry6Aa杀虫多肽或核苷酸代表小于约30％、小于约20％、小于约10％、小于约5％、小于约4％、小于约3％或小于约2％、或小于约1％(按干重计)的分离的工程化的Cry6Aa杀虫多肽或核苷酸。

Cry6Aa和变体因为其效力和独特的序列以及结构同一性而是Cry34Ab1/Cry35Ab和其他玉米根虫杀虫剂的潜在金字塔配偶体。此外，它不与Cry34Ab1/Cry35Ab1或Cry3Aa竞争WCR刷状缘膜囊泡上的结合位点(Li等人，2013)。在胞质溶胶中表达Cry6Aa的转基因植物不是有效的；分析显示蛋白质易受蛋白水解加工和可能的失活的影响，类似于实施例3中所述。限制蛋白水解的一种策略是如实施例1中所述地将重组蛋白引导到亚细胞隔室(由Benchabane，2008综述)。限制蛋白水解和可能失活的另一种策略是通过蛋白质工程化。

工程化的Cry6Aa杀虫毒素.本发明涵盖具有杀虫活性的工程化的Cry6Aa及其变体。通过术语“变体”，申请人意图包括片段、某些缺失和插入突变体、取代以及某些融合或嵌合蛋白。

本发明包括具有与SEQ ID NO:2的氨基酸1至47590％、91％、92％、93、94％、95％、96％、97％、98％或99％同一的毒素区段的DIG-177杀虫蛋白变体。可以通过制备随机突变体来制备变体或者可以设计变体。在设计的突变体的情况下，当维持在负责生物活性或者参与确定最终负责生物活性的三维构型的毒素关键区域中的氨基酸同一性时，存在产生具有与天然毒素相似活性的变体的高概率。如果取代是保守的，也将存在保留活性的高概率。氨基酸可以置于以下类别中：非极性、不带电荷的极性、碱性和酸性。一种类型氨基酸被相同类型的另一氨基酸替代所借助于的保守取代最不可能实质上改变变体的生物活性。表1提供了属于每个类别的氨基酸的实例的列表。

表1.氨基酸的生物化学类别

在一些情况下，也可以进行非保守取代。关键因素是这些取代必须不会显著减损毒素的生物活性。变体包括由于诱变而在氨基酸序列上不同的多肽。本发明涵盖的变体蛋白是具有生物活性的，即它们继续具有天然蛋白质的所需生物活性，即保留杀虫活性。

变体蛋白质也可以被设计成在序列水平上不同，但保留相同或相似的整体基本三维结构、表面电荷分布等。参见例如美国专利No.7058515；Larson等人，(2002)；Stemmer(1994a,1994b,1995)和Crameri等人.(1996a,1996b,1997)。美国专利No US 8,513,492B2.

核酸.编码工程化的Cry6Aa杀虫毒素的分离核酸及其补体是本发明的一个方面。术语“分离的”在本文中如上定义。由于遗传密码的冗余，各种不同的DNA序列可以编码本文公开的氨基酸序列。这完全在本领域中受过训练的人员用以产生编码相同或基本上相同的杀虫蛋白的这些替代DNA序列的技能范围内。

基因合成.编码本文所述的工程化的Cry6Aa杀虫蛋白的基因可以通过在本领域中众所周知的多种方法制备。例如，合成基因区段和合成基因可以通过亚磷酸三酯和亚磷酰胺化学制备(Caruthers等人，1987)，并且商业供应商可用于根据需要进行基因合成。全长基因可以以多种方式装配，包括例如通过连接限制性片段或重叠寡核苷酸的聚合酶链式反应装配(Stewart和Burgin，2005)。此外，末端基因缺失和添加可以通过使用位点特异性末端寡核苷酸的PCR扩增来进行。

编码工程化的Cry6Aa杀虫蛋白的核酸可以例如通过由若干个商业供应商中的任何一个目前实践的方法进行的合成构建来制备。(例如美国专利No.7,482,119)。这些基因或其部分或变体也可以合成地构建，例如通过使用基因合成仪和例如美国专利No.5,380,831的设计方法。或者，可以使用用于制备点突变、添加和/或缺失的标准分子生物学技术轻易地构建合成或天然存在的基因的变体。这些基因的片段也可以根据标准程序使用市售的外切核酸酶或内切核酸酶制备。例如，诸如Bal31或定点诱变的酶可用于从这些基因的末端系统地切割核苷酸。此外，可以使用多种限制酶获得编码活性毒素片段的基因片段。

在给定工程化的Cry6Aa杀虫蛋白的氨基酸序列的情况下，可以通过以下方式来设计编码序列：使用预期宿主首选的同义密码子逆向翻译编码序列，然后使用替代的同义密码子来纯化序列以去除可能引起转录、翻译或mRNA稳定性问题的序列的序列。此外，可以使用同义密码子在非工程化的Cry6Aa阅读框(即，阅读框2、3、4、5和6)中引入终止密码子以消除假长开放阅读框。

定量多肽或核酸序列同一性.通过首先出于最佳比较目的比对序列来确定两个氨基酸序列或两个核酸序列的同一性百分比。两个序列之间的同一性百分比是由序列共享的相同位置的数目的函数(即，同一性百分比＝相同位置的数目/位置的总数目(例如重叠位置)×100)。在一个实施方案中，两个序列具有相同的长度。两个序列之间的同一性百分比可以使用类似于下面描述的技术在允许或不允许空位的情况下来确定。在计算同一性百分比时，通常计数精确匹配。

可以使用数学算法来实现对两个序列之间的同一性百分比的确定。这种算法的非限制性实例是Altschul等人，(1990)，以及Karlin和Altschul(1990)的算法，在Karlin和Altschul(1993)中经改进，并且被结合到BLASTN和BLASTX程序中。BLAST搜索可以方便地用于鉴定与核酸或蛋白质数据库中的查询序列同源(相似)的序列。可以进行BLASTN搜索(得分＝100，字长度＝12)以鉴定与所要求保护的本发明的核酸分子具有同源性的核苷酸序列。可以进行BLASTX搜索(得分＝50，字长度＝3)以鉴定与所要求保护的本发明的杀虫蛋白分子具有同源性的氨基酸序列。

可以利用空位BLAST(Altschul等人，1997)获得空位比对以用于比较目的。或者，PSI-Blast可用于进行迭代检索，其检测分子间的距离关系(Altschul等人，1997)。当利用BLAST、空位BLAST和PSI-Blast程序时，可使用各个程序的缺省参数。参见www.ncbi.nlm.nih.gov。

用于比较序列的数学算法的非限制性实例是ClustalW算法(Thompson等人，1994)。ClustalW比较序列并比对整个氨基酸或DNA序列，并因此可提供有关整个氨基酸序列或核苷酸序列的序列保守性的数据。ClustalW算法在若干个市售DNA/氨基酸分析软件包中使用，诸如Vector NTI Program Suite(英杰公司(Invitrogen，Inc.)，新墨西哥州卡尔斯巴德)的ALIGNX模块。当用ALIGNX比对氨基酸序列时，可方便地使用缺省设置，即10的空位开放罚分、0.1空位延伸罚分以及blosum63mt2比较矩阵，从而评估两个序列间的氨基酸类似性(共有)或同一性百分比。当使用ALIGNX比对DNA序列时，可方便地使用缺省设置，即15的空位开放罚分、6.6的空位延伸罚分以及swgapdnamt比较矩阵，以评估两个序列间的同一性百分比。

用于比较序列的数学算法的另一非限制性实例是Myers和Miller(1988)的算法。这种算法已整合至wSTRETCHER程序中，该程序是wEMBOSS序列比对软件包(可在http://emboss.sourceforge.net/中获得)的一部分。wSTRETCHER使用标准动态编程算法(其使用线性空间)的修正版计算两个序列的最佳全局比对。用于计算比对的取代矩阵、空位插入罚分和空位延伸罚分可以具体指定。当利用wSTRETCHER程序比较核苷酸序列时，可与打分矩阵文件EDNAFULL一起使用为16的空位开放罚分和为4的空位延伸罚分。当用于比较氨基酸序列时，可与EBLOSUM62打分矩阵文件一起使用为12的空位开放罚分和为2的空位延伸罚分。

用于比较序列的数学算法的其他非限制性实例是Needleman和Wunsch(1970)的算法，其整合至序列比对软件包GAP版本10和wNEEDLE(http://emboss.sourceforge.net/)中。GAP版本10可使用以下参数从而用于确定序列同一性或相似性：对于核苷酸序列，使用为50的空位权重(GAP Weight)和为3的长度权重(Length Weight)，以及nwsgapdna.cmp打分矩阵，建立同一性％和相似性％。对于氨基酸序列比较，使用为8的空位权重和为2的长度权重，以及BLOSUM62打分程序，建立同一性％或相似性％。

wNEEDLE读取两个输入序列，沿着它们的全长找出最佳比对(包括空位)，并在文件中写入它们的最佳全局序列比对。该算法使用含有每一种可能的残基或核苷酸匹配的值的打分矩阵，探索所有可能的比对并选择最佳的。wNEEDLE寻找具有最大可能得分的比对，其中比对的得分等于获自打分矩阵的匹配总分减去从比对序列中的开放和延伸空位产生的罚分。取代矩阵和空位开放及延伸罚分是用户指定的。当比较氨基酸序列时，使用为10的缺省空位开放罚分、为0.5的空位延伸罚分以及EBLOSUM62比较矩阵。当使用wNEEDLE比较DNA序列时，使用为10的缺省空位开放罚分、为0.5的空位延伸罚分以及EDNAFULL比较矩阵。

也可使用等效程序。“等同程序”意指任何序列比较程序，其为所考虑的任何两个序列产生这样的比对，当与由ALIGNX、wNEEDLE或wSTRETCHER产生的对应比对相比较时，该比对具有相同的核苷酸或氨基酸残基匹配和相同的序列同一性百分比。该同一性％是两个序列间同一的匹配除以报告的比对区(包括长度上的任何空位)的百分比，以及相似性％是两个序列间的匹配除以报告的比对区(包括长度上的任何空位)的百分比。

也可通过检查手工地进行比对。

重组宿主.可以将本发明的杀虫蛋白编码基因导入多种微生物、真菌或植物宿主中。杀虫蛋白基因的表达直接或间接导致杀虫蛋白的细胞内产生和保持。使用合适的微生物宿主，例如假单胞菌属(Pseudomonas)，可将微生物施用于害虫的环境中，微生物可以在该环境中增殖并且被摄食。结果是害虫的控制。或者，可以在延长该蛋白的活性和稳定重组宿主细胞的条件下处理含有杀虫蛋白基因的微生物。经处理的细胞，其包含保留杀虫活性的本发明的经处理杀虫多肽，可以应用于目标害虫的环境以控制害虫。

当通过合适的DNA构建体，例如载体将B.t.杀虫蛋白基因导入微生物宿主，并且将所述宿主以活的状态应用于环境时，使用某些宿主微生物是必须的。选择已知占据一种或多种所关注的作物的“植物圈”(叶面、叶圈、根际和/或根面)的微生物宿主。选择这些微生物以便能够在特定环境(作物和其他昆虫栖息地)与野生型本土微生物成功地竞争，提供表达多肽杀虫剂的基因的稳定保持和表达，以及如希望的那样，提供对杀虫剂的改进保护以免于环境降解和失活。

已知大量微生物栖息在多种重要作物的叶面(植物叶子的表面)和/或根际(植物根周围的土壤)。这些微生物包括细菌、藻类和真菌。尤其受关注的是微生物，诸如细菌，例如假单胞菌属(Pseudomonas)、欧文氏菌属(Erwinia)、沙雷氏菌属(Serratia)、克雷伯氏菌属(Klebsiella)、黄单胞菌属(Xanthomonas)、链霉菌属(Streptomyces)、根瘤菌属(Rhizobium)、中华根瘤菌属(Sinorhizobium)、红假单胞菌属(Rhodopseudomonas)、嗜甲基菌属(Methylophilius)、土壤杆菌属(Agrobacterium)、醋杆菌属(Acetobacter)、乳杆菌属(Lactobacillus)、节杆菌属(Arthrobacter)、固氮菌属(Azotobacter)、明串珠菌属(Leuconostoc)、和产碱菌属(Alcaligenes)。尤其受关注的是诸如以下的植物圈细菌物种：丁香假单胞菌(Pseudomonas syringae)、荧光假单胞菌(Pseudomonas fluorescens)、粘质沙雷氏菌(Serratia marcescens)、木醋杆菌(Acetobacter xylinum)、根瘤土壤杆菌(Agrobacterium tumefaciens)、放射形土壤杆菌(Agrobacterium radiobacter)、球形红假单胞菌(Rhodopseudomonas spheroides)、野油菜黄单胞菌(Xanthomonas campestris)、苜蓿根瘤菌(Sinorhizobiummeliloti)(以前为Rhizobium meliloti)、真养产碱菌(Alcaligenes eutrophus)、和棕色固氮菌(Azotobacter vinelandii)。进一步受关注的是真菌，尤其是酵母，例如酵母属(Saccharomyces)、隐球酵母属(Cryptococcus)、克鲁维酵母属(Kluyveromyces)、掷孢酵母属(Sporobolomyces)、红酵母属(Rhodotorula)和短梗霉属(Aureobasidium)，并且尤其受关注的是以及诸如以下的植物圈酵母物种：深红酵母(Rhodotorula rubra)、红酵母(R.glutinis)、海滨红酵母(R.marina)、橙黄红酵母(R.aurantiaca)、浅白隐球酵母(Cryptococcus albidus)、流散隐球酵母(C.diffluens)、变黄罗伦隐球酵母(C.laurentii)、罗斯酵母(Saccharomyces rosei)、普地酵母(S.pretoriensis)、酿酒酵母(S.cerevisiae)、掷孢酵母(Sporobolomyces roseus)、香气掷孢酵母(S.odorus)、佛地克鲁维酵母(Kluyveromyces veronae)和出芽短梗霉(Aureobasidium pollulans)。特别受关注的是有色的微生物。

分离的毒素多肽和本发明的组合物.可以处理或制备本发明的工程化的Cry6Aa虫毒素多肽，例如以制备配制的杀虫剂组合物。配制的杀虫剂组合物的实例包括蛋白质组合物、可喷雾蛋白质组合物、诱饵基质，或其他递送系统。在一个实例中，表达本发明的工程化的Cry6Aa杀虫蛋白的B.t.细胞或重组宿主细胞使用标准的技术培养基和发酵技术培养。一旦发酵周期完成，可以通过本领域已知的方法从发酵液分离B.t.孢子或其他重组宿主细胞和/或杀虫蛋白晶体。B.t.孢子或重组宿主细胞也可以在施用或配制施用于植物之前进行处理。例如，可以化学处理分离的B.t.孢子和/或毒素晶体以延长杀虫活性，由此包括本发明的经处理的多肽。在重组宿主中生长B.t.杀虫多肽，然后处理B.t.以延长杀虫活性的方法是已知的并已公开。参见例如美国专利No.4,695,462和No.4,695,455，以及Gaertner等人，1993。

本发明的分离或处理的工程化的Cry6Aa杀虫蛋白可以配制成细碎的颗粒状固体颗粒、小丸，可湿性粉剂，粉尘(dust)、水悬浮液或分散体、乳液、喷雾剂、液体浓缩物，或其他杀虫剂制剂。这些杀虫剂制剂通过将本文的工程化的Cry6Aa杀虫剂多肽与佐剂、稀释剂、表面活性剂、分散剂、惰性载体和其他组分组合来制备，以便于处理和应用以控制一种或多种目标害虫。此类制剂成分是本领域已知的，施用方法和确定提供期望的杀虫活性的B.t.孢子和/或分离的工程化的Cry6Aa多肽晶体的水平的方法也是本领域已知的。

控制害虫的方法..当昆虫摄入通过杀虫剂组合物(例如配制的蛋白质组合物、可喷雾的蛋白质组合物、诱饵基质、转基因植物表达、或者另一种递送系统)传递的有效量的本文公开的＝工程化的Cry6Aa杀虫蛋白时，结果通常是昆虫死亡，或昆虫不再摄食给昆虫提供该杀虫蛋白质的来源。本文中的术语“传递的”或“传递”意在包括在植物组织上或在植物组织中放置或提供杀虫有效量的蛋白质毒素的任何方法。这包括在植物细胞中表达杀虫蛋白，或对植物的任何表面施用有效量。

可以多种方式“施用”或提供主题蛋白昆虫毒素与靶标昆虫接触。例如，本发明的工程化的Cry6Aa杀虫蛋白可以在与佐剂、稀释剂、载体等等一起配制以提供细碎固体颗粒物、颗粒、小丸、可湿性粉剂、粉尘、水悬液或分散体、以及乳液形式的组合物之后施用。或者，可通过转基因植物(其中蛋白质由该植物表达，并存在于该植物中)来递送工程化的Cry6Aa杀虫多肽，此为本领域熟知的。还可选择性地实现毒素基因在植物特定组织(诸如根、叶等)中的表达。这例如可通过使用组织特异性启动子而实现。喷洒施用是另一实例，并且也是本领域中已知的。可合适地配制该主题蛋白质用于预期的终端用途，并且然后喷雾(或以其他方式施用)至植物和/或植物周围/至待保护的植物附近——在发现侵染之前、发现靶标昆虫之后、之前及之后等。例如，还可使用诱饵颗粒，并且这也是本领域中已知的。

转基因植物.本文中公开的工程化的Cry6Aa杀虫蛋白可用于实际地保护任何类型的植物免遭害虫的损害。此类植物的非限制性实例包括马铃薯、茄子、番茄、辣椒、烟草，以及其他茄科植物。此类植物的其他实例包括玉蜀黍、向日葵、大豆、低芥酸菜子、稻、高粱、小麦、大麦、蔬菜、观赏植物、辣椒(包括小辣椒)、甜菜、果树以及草皮，仅略举数例。转化植物的方法是本领域中熟知的，并且在实施例中描述了例示性的转化方法。

本发明的一个优选实施方案是用编码工程化的Cry6Aa杀虫蛋白及其变体的基因在遗传上转化植物。经转化的植物由于在该转化植物的细胞中存在控制量的该主题杀虫蛋白或其变体，因此能耐受昆虫靶标害虫的攻击。通过结合编码本发明的杀虫蛋白的功能性遗传材料，成虫或幼虫在食用含有所要求保护的毒素的植物组织后将死亡。已转化了单子叶和双子叶分类的大量成员。转基因农作物以及果树和蔬菜是商业上感兴趣的。此类作物包括但不限于玉蜀黍、稻、大豆、低芥酸菜子、向日葵、紫花苜蓿、高粱、小麦、棉花、花生、番茄、马铃薯等。更优选的一组作物是玉蜀黍，大豆和棉花。最优选的作物是玉蜀黍。

存在若干种将外源遗传材料导入植物细胞，以及获得稳定保持并表达该引入的基因的技术。此类技术包括将包被在微粒上的遗传物质直接导入(acceleration)细胞中(美国专利No.4945050和美国专利No.5141131)。可以使用土壤杆菌(Agrobacterium)技术转化植物，参见美国专利No.5177010、欧洲专利No.EP131624B1、欧洲专利No.EP159418B1、欧洲专利No.EP176112B1、美国专利No.5149645、EP120516B1、美国专利No.5464763、美国专利No.4693976、欧洲专利No.EP116718B1、欧洲专利No.EP290799B1、欧洲专利No.EP320500B1、欧洲专利No.EP604662B1、美国专利No.7060876、美国专利No.6037526、美国专利No.6376234、欧洲专利No.EP292435B1、美国专利No.5231019、美国专利No.5463174、美国专利No.4762785、美国专利No.5608142，以及美国专利No.5159135。其他转化技术包括WHISKERS^TM技术，参见美国专利No.5302523和美国专利No.5464765。电穿孔技术也已被用于转化植物，参见WO1987006614、美国专利No.5472869、美国专利No.5384253、WO199209696、美国专利No.6074877、WO1993021335，和美国专利No.5679558。除了用于转化植物的多种技术之外，与外源基因接触的组织类型也可以不同。此类组织将包括但不限于胚胎组织、I型和II型愈伤组织、胚轴、分生组织等。应用本领域技术人员技术范围内的合适技术，几乎所有植物组织均可在分化期间转化。

编码工程化的Cry6Aa杀虫蛋白的基因可使用各种如上公开的本领域熟知的技术插入植物细胞中。例如，包含在大肠杆菌(Escherichia coli)中起作用的允许对转化微生物细胞进行选择的标记物和复制系统的大量克隆载体可用于制备和修饰用于插入到高等植物中的外来基因。此类操纵可包括例如，插入突变、截短、添加或取代，如预期用途所期望的那样。这些载体包括例如pBR322、pUC系列、M13mp系列、pACYC184等。因此，可以将编码本发明的蛋白毒素或其变体的序列插入到载体的适当限制性位点。所得到的质粒用于转化大肠杆菌，将得到的大肠杆菌细胞在适当的营养培养基中进行培养，然后收获并裂解，从而回收可工作量的质粒。通常实施序列分析、限制性片段分析、电泳和其他生物化学-分子生物学方法作为分析方法。每一次操纵之后，可以切割所使用的DNA序列并且连接到下一个DNA序列中。可以将每一个操纵的DNA序列克隆到相同的或者其他的质粒中。

对于含T-DNA的载体用于转化植物细胞的用途已经得到广泛研究并在欧洲专利No.EP120516B1；Lee和Gelvin(2008)；Fraley等人(1986)以及An等人(1985)中得到充分地叙述，并在田间良好建立。

一旦插入的DNA被整合进入植物基因组，则它在随后的世代中是相对稳定的。用于转化植物细胞的载体通常含有编码赋予转化的植物细胞除草剂耐性或者抗生素(诸如草丁膦、双丙氨磷、卡那霉素、新霉素、G418、博来霉素、潮霉素)抗性的蛋白质的选择标记基因，或编码对草甘膦、甲氨蝶呤、吲哚啉酮、磺酰脲和三唑并嘧啶类除草剂，诸如氯磺隆、溴苯腈、茅草枯等的耐性的基因。进一步受关注的是赋予对除草剂如盖草能、喹禾灵、禾草灵等的耐性的基因，例如AAD基因(美国专利申请No.20090093366)。单独采用的选择标记基因因此应当允许选择经转化的细胞，同时通过选择性化合物抑制不含插入DNA的细胞的生长。

大量的技术可用于将DNA插入到宿主植物细胞中。这些技术包括使用由作为转化剂的根癌农杆菌(Agrobacterium tumefaciens)或发根农杆菌(Agrobacteriumrhizogenes)传递的T-DNA进行转化。此外，也可采用植物原生质体与含有待传递DNA的脂质体的融合、直接注射DNA、基因枪转化(微粒轰击)或电穿孔，以及其他可能的方法。

在本发明一个优选实施方案中，用其中已经针对植物优化了蛋白质编码区域的密码子选择的基因转化植物。参见例如美国专利No.5,380,831。例如，本发明的工程化的Cry6Aa杀虫毒素可以经优化以用于在双子叶植物如马铃薯、茄子、番茄、辣椒、烟草或其他茄科植物中表达。本发明的工程化的Cry6Aa杀虫毒素也可以优化用于在其他双子叶植物如大豆和棉花中表达，或在单子叶植物如玉蜀黍(Zea mays)(玉米)中表达。此外，可以有利地使用编码截短毒素的植物。截短的毒素通常将编码全长毒素的约55％至约80％。产生用于植物中的合成B.t.基因的方法是本领域中已知的(Stewart，2007)。

无论转化技术如何，优选将该基因掺入通过在载体中包含植物启动子从而适于在植物细胞中表达B.t.杀虫毒素基因和变体的基因转化载体中。除了植物启动子之外，可在植物细胞中有效地使用来自各种来源的启动子以表达外源基因。例如，可使用细菌来源的启动子，诸如章鱼碱合酶启动子、章鱼碱合酶启动子、甘露碱合酶启动子；可以使用病毒来源的启动子，诸如花椰菜花叶病毒(CaMV)的35S和19S启动子等等。植物衍生的启动子包括但不限于核酮糖-1,6-二磷酸(RUBP)羧化酶小亚基(ssu)、β-伴大豆球蛋白启动子、菜豆蛋白启动子、ADH(醇脱氢酶)启动子、热休克启动子、ADF(肌动蛋白解聚因子)启动子和组织特异性启动子。启动子还可含有可改善转录效率的某些增强子序列元件。典型的增强子包括但不限于ADH1-内含子1和ADH1-内含子6。可使用组成型启动子。组成型启动子在几乎所有的细胞类型并且在几乎所有的时间引导持续的基因表达(例如，肌动蛋白、遍在蛋白、CaMV35S)。组织特异性启动子负责特定细胞或组织类型中的基因表达，诸如叶子或种子(例如，玉米蛋白、油质蛋白、油菜籽蛋白、ACP(酰基载体蛋白))，也可使用这些启动子。也可使用在植物发育的某一阶段活化，以及在特定植物组织及器官中活化的启动子。此类启动子的实例包括但不限于根特异性、花粉特异性、胚特异性、玉米穗丝特异性、棉纤维特异性、种子胚乳特异性、韧皮部特异性等等的启动子。

在某些条件下，可能期望使用可诱导的启动子。可诱导启动子负责响应特定信号而表达，诸如：物理刺激(例如，热休克基因)；光(例如，RUBP羧化酶)；激素(例如，糖皮质激素)；抗生素(例如，四环素)；代谢物；以及胁迫(例如，干旱)。可使用在植物中起作用的其他期望的转录和翻译元件，诸如5'非翻译引导序列、RNA转录终止序列和聚腺苷酸附加信号序列。多种植物特异性基因转运载体是本领域中已知的。

靶害虫.本发明的工程化的Cry6Aa杀虫蛋白特别适合用于控制害虫。鞘翅目昆虫是一群重要的农业、园艺和家庭害虫，每年造成极大量的损失。这一昆虫大目涵盖食叶和食根的幼虫和成虫，包括例如金花虫科(Chrysomelidae)、瓢虫科(Coccinellidae)、象鼻虫科(Curculionidae)、皮蠹科(Dermestidae)、叩甲科(Elateridae)、金龟子科(Scarabaeidae)、小蠹科(Scolytidae)、和拟步甲科(Tenebrionidae)等昆虫科的成员。这些科中包括金花虫科中的金花虫(leaf beetles)和潜叶虫(leaf miners)，马铃薯甲虫(例如科罗拉多马铃薯甲虫(Leptinotarsa decemlineata Say)、葡萄肖叶甲(Colaspisbrunnea Fabricius)、黑角负泥虫(Oulemamelanopus Linnaeus)、向日葵叶甲(Zygogrammaexclamationis Fabricius))，和瓢虫科的甲虫(例如墨西哥豆瓢虫(Epilachnavarivestis Mulsant))。进一步的实例是金龟子(chafer)和金龟子科的其他甲虫(例如日本甲虫(Popillia japonica Newman)、北方圆头犀金龟(白蛴螬，Cyclocephala borealisArrow)、南方圆头犀金龟(白蛴螬，Cyclocephala immaculata Olivier)、欧洲金龟子(Rhizotrogus majalis Razoumowsky)、白蛴螬(Phyllophaga crinita Burmeister)、胡萝卜甲虫(Ligyrus gibbosus De Geer)，以及丝爪鳃金龟属(Holotrichia spp)和鳃角金龟属(Melolontha spp.)的金龟子)。鞘翅目昆虫的更多实例有象鼻虫(例如苜蓿叶象甲(Anthonomus grandis Boheman)、稻水象甲(Lissorhoptrus oryzophilus Kuschel)、谷象(Sitophilus grananus Linnaeus)、米象(Sitophilus oryzae Linnaeus)、和三叶草叶象(Hyperapunctata Fabricius))。还包括玉米象虫(Sphenophorus maidis Chittenden)、跳甲(例如玉米跳甲(Chaetocnema pulicara Melsheimer)和十字花科跳甲(Phyllotretacruciferae Goeze))、十一星瓜叶甲(Diabrotica undecimpunctata)、和根虫(例如西方玉米根虫(Diabrotica virgifera virgifera LeConte)、北方玉米根虫(Diabrotica barbenSmith&Lawrence)、南方玉米根虫(Diabroticaundecimpunctata howardi Barber)、墨西哥玉米根虫(D.virgifera zeae Krysan and Smith)、黄瓜条根萤叶甲(D.balteataLeConte)、柔嫩黄瓜十一星叶甲(D.undecimpunctata tenella)、南美叶甲(D.speciosaGermar)和甘薯十一星叶甲(D.u.undecimpunctata Mannerheim)。鞘翅目害虫的更多实例有丽金龟亚科(Rutelinae)的甲虫(丽金龟)，诸如异丽金龟属(Anomala)(包括A.marginata、A.lucicola、A.oblivia和A.orientalis)。其他鞘翅目昆虫有皮蠹科的地毯甲虫、来自叩甲科的金针虫(例如梳爪叩甲属(Melanotus spp.)、单叶叩甲属(Conoderusspp.)、丘胸叩甲属(Limonius spp.)、细胸叩甲属(Agriotes spp.)、金针虫属(Cteniceraspp.)、Aeolus spp.))、来自小蠹科的树皮甲虫(bark beetles)、来自拟步甲科的甲虫(例如伪金针虫属Eleodes spp)。上文列出的任何属(以及其他属)，一般而言，也可以作为包含单独的工程化的Cry6Aa杀虫多肽或其与另一杀虫剂的组合的杀虫组合物的靶标而作为本主题发明的一部分。这些属中的任何者的任何其他昆虫(作为靶标)也包括在本发明的范围内。

考虑了工程化的Cry6Aa杀虫蛋白用于控制作物植物的鞘翅目害虫的用途。在一些实施方案中，可以经济地部署Cry蛋白用于控制害虫，这些害虫包括但不限于例如，根虫如西方玉米根虫(Diabrotica virgifera virgifera LeConte)、北方玉米根虫(Diabroticabarberi Smith&Lawrence)、南方玉米根虫(Diabroticaundecimpunctata howardiBarber)、墨西哥玉米根虫(D.virgifera zeae Krysan and Smith)、黄瓜条根萤叶甲(D.balteata LeConte)、柔嫩黄瓜十一星叶甲(D.undecimpunctata tenella)、南美叶甲(D.speciosa Germar)和甘薯十一星叶甲(D.u.undecimpunctata Mannerheim)，以及蛴螬如北方圆头犀金龟(Cyclocephala borealis)、圆头无斑犀金龟(Cyclocephalaimmaculate)(南方圆头犀金龟)和日本丽金龟()(日本甲虫)的幼虫。

鳞翅目是另一组重要的农业、园艺和家庭害虫，每年造成极大量的损失。本发明提供工程化的Cry6Aa杀虫蛋白与其他杀虫剂组合用于控制该目中的害虫的用途。这一目昆虫涵盖食叶和食根的幼虫和成虫，包括例如灯蛾科(Arctiidae)、麦蛾科(Gelechiidae)、尺蛾科(Geometridae)、枯叶蛾科(Lasiocampidae)、毒蛾科(Lymantriidae)、夜蛾科(Noctuidae)、螟蛾科(Pyralidae)、透翅蛾科(Sesiidae)、天蛾科(Sphingidae)、谷蛾科(Tineidae)、和卷叶蛾科(Tortricidae)等昆虫科的成员。鳞翅目害虫包括但不限于：小蜡螟(Achoroia grisella)、东部黑头长翅卷蛾(Acleris gloverana)、黑头长翅卷蛾(Acleris variana)、棉褐带卷叶蛾(Adoxophyes orana)、小地老虎(Agrotis ipsilon)、棉叶波纹夜蛾(Alabama argillacea)、秋星尺蠖(Alsophila pometaria)、脐橙螟(Amyeloistransitella)、地中海粉螟(Anagasta kuehniella)、桃条麦蛾(Anarsia lineatella)、栎黄条大香蛾(Anisota senatoria)、柞蚕(Antheraea pernyi)、黎豆夜蛾(Anticarsiagemmatalis)、黄卷蛾属(Archips sp.)、条卷蛾属(Argyrotaenia sp.)、粗皮夜蛾(Athetismindara)、家蚕(Bombyx mori)、棉潜叶蛾(Bucculatrix thurberiella)、粉斑螟(Cadracautella)、色卷蛾属(Choristoneura sp.)、Cochylls hospes、苜蓿粉蝶(Coliaseurytheme)、米蛾(Corcyra cephalonica)、Cydia latiferreanus、苹果蠹蛾(Cydiapomonella)、核桃配片舟蛾(Datana integerrima)、落叶松毛虫(Dendrolimussibericus)、葡萄小卷叶野螟(Desmi afeneralis)、甜瓜绢野螟(Diaphania hyalinata)、黄瓜绢野螟(Diaphania nitidalis)、西南玉米杆草螟(Diatraea grandiosella)(巨座玉米螟)、小蔗螟(Diatraea saccharalis)(甘蔗螟)、白尺蠖蛾(Ennomos subsignaria)、蛀茎螟蛾(Eoreuma loftini)、烟草粉螟(Esphestia elutella)、菩提松尺蠖(Erannistilaria)、细斑灯蛾(Estigmene acrea)、Eulia salubricola、Eupocoellia ambiguella、女贞细卷蛾(Eupoecilia ambiguella)、黄毒蛾(Euproctis chrysorrhoea)、暗缘地老虎(Euxoa messoria)、蜡螟(Galleria mellonella)、梨小食心虫(Grapholita molesta)、黑拟蛉蛾(Harrisina americana)、棉花暝铃(Helicoverpa subflexa)、谷实夜蛾(Helicoverpa zea)(玉米夜蛾)、烟芽夜蛾(Heliothis virescens)(烟草夜蛾)、新墨西哥草原毛虫(Hemileuca oliviae)、向日葵斑螟(Homoeosoma electellum)、美国白蛾(Hyphantia cunea)、番前蠹蛾(Keiferial ycopersicella)、铁杉尺蠖(Lambdinafiscellaria fiscellaria)、Lambdina fiscellaria lugubrosa、雪毒蛾(Leucomasalicis)、葡萄浆果小卷蛾(Lobesia botrana)、西部豆类切根虫(Loxagrotis albicosta)(西部豆地老虎)、草地螟(Loxostege sticticalis)、舞毒蛾(Lymantria dispar)、锄须丛螟(Macalla thyrisalis)、天幕毛虫属(Malacosoma sp.)、甘蓝灯蛾(Mamestrabrassicae)、蓓带夜蛾(Mamestra configurata)、番茄天蛾(Manduca quinquemaculata)、烟草天蛾(Manduca sexta)、豆野螟(Maruca testulalis)、斑马纹夜蛾(Melanchrapicta)、秋尺蛾(Operophtera brumata)、古毒蛾属(Orgyia sp.)、欧洲玉米螟(Ostrinianubilalis)(European corn borer)、春尺蠖(Paleacrita vernata)、驻茎夜蛾(Papiapemanebris)(普通钻心虫)、))、美洲大芷凤蝶(Papilio cresphontes)、棉红铃虫(Pectinophora gossypiella)、加州懈蛾(Phryganidia californica)、斑幕潜叶蛾(Phyllonorycter blancardella)、暗脉菜粉蝶(Pieris napi)、油菜菜粉蝶(Pierisrapae)、苜蓿绿夜蛾(Plathypena scabra)、Platynota flouendana、荷兰石竹小卷蛾(Platynota stultana)、洋葱羽蛾(Platyptilia carduidactyla)、印度谷螟(Plodiainterpunctella)、菱纹背蛾(Plutella xylostella)(小菜蛾)、美国纹白蝶(Pontiaprotodice)、一点粘虫(Pseudaletia unipuncta)(粘虫)、、大豆夜蛾(Pseudoplasiaincludens)、杂食尺蠖(Sabulodes aegrotata)、红山背舟蛾(Schizura concinna)、麦蛾(Sitotroga cerealella)、苹白小卷蛾(Spilonta ocellana)、草地贪夜蛾(Spodopterafrugiperda)(秋粘虫)、甜菜夜蛾(Spodoptera exigua)(甜菜粘虫)、松异舟蛾(Thaurnstopoea pityocampa)、Ensola bisselliella、粉纹夜蛾(Trichoplusia ni)(甘蓝银纹夜蛾)、温室结网野螟(Udea rubigalis)、Xylomyges curiails和苹果巢蛾(Yponomeuta padella)。

还考虑了工程化的Cry6Aa杀虫蛋白用于控制寄生线虫动物的用途，所述寄生线虫动物包括但不限于：根瘤线虫(Meloidogyne incognita)和大豆胞囊线虫(Heteroderaglycines)。

抗毒素抗体.可使用本领域的标准方法容易地制备针对本文公开的毒素、或针对这些毒素的等价毒素或片段的抗体。此类抗体可用于纯化或检测工程化的Cry6Aa毒素的存在。

一旦分离了B.t.杀虫蛋白，则可通过本领域熟知的传统方法产生对该蛋白特异的抗体。在数周或数月的期间内重复注射至所选择的宿主将诱发免疫反应，并产生显著的抗B.t.毒素血清滴度。优选的宿主是哺乳动物物种，更优选的物种是兔、山羊、绵羊和小鼠。从此类免疫动物获取的血液可通过已建立的方法加工，以获得与B.t.杀虫毒素反应的抗血清(多克隆抗体)。然后可根据本领域已知的技术，通过吸附至该毒素从而亲和纯化该抗血清。亲和纯化的抗血清可通过使用本领域已知的方法分离该抗血清内的免疫球蛋白级分从而进一步加以纯化。得到的材料将是与B.t.杀虫毒素反应的免疫球蛋白的异质群体。

抗B.t.毒素抗体还可通过制备半合成免疫原而产生，该半合成免疫原由与免疫原性载体缀合的B.t.杀虫毒素的合成肽片段组成。可用于制备肽片段的多种方案和仪器是本领域熟知的。许多合适的免疫原性载体诸如牛血清白蛋白或钥孔戚血蓝蛋白也是本领域熟知的，用于偶联免疫原和载体蛋白的技术也是熟知的。一旦构建了半合成免疫原，则制备对B.t.杀虫毒素片段特异的抗体的方法与制备和天然B.t.毒素反应的抗体的方法是相同的。

抗B.t.毒素单克隆抗体(MAb)可容易地使用纯化的B.t.杀虫蛋白制备。用于产生MAb的方法已实践20多年，并且是本领域普通技术人员熟知的。重复腹膜内或皮下注射佐剂中的纯化B.t.杀虫蛋白将在多数动物中诱发免疫反应。从动物中取出高免疫B-淋巴细胞，并与能够无限培养的合适的融合配偶体细胞系融合。B淋巴细胞可被高免疫并可用于产生MAb的优选动物是哺乳动物。更优选的动物是大鼠和小鼠，并且最优选的是BALB/c小鼠品种。

许多哺乳动物细胞系是用于产生杂交瘤细胞的合适融合配偶体。许多此类细胞系可从美国典型培养物保藏中心(ATCC，弗吉尼亚州马纳萨斯)及商业供应商处获得。优选的融合配偶体细胞系来源于小鼠骨髓瘤，并且骨髓瘤-653细胞系(Ventrex公司，缅因州波特兰)是最优选的。一旦融合，则在选择生长培养基中培养所得杂交瘤细胞一周至两周。两个熟知的选择系统可用于从混合的杂交瘤培养物中消除未融合的骨髓瘤细胞、或骨髓瘤细胞之间的融合。选择系统的选择取决于免疫的小鼠品种以及所使用的杂交瘤融合配偶体。可使用Taggart和Samloff(1983)描述的AAT选择系统；然而，由Littlefield(1964)描述的HAT(次黄嘌呤、氨基蝶呤、胸腺嘧啶)选择系统是优选的，因为其可与上述优选的小鼠品种及融合配偶体相容。然后从用过的生长培养基中筛选免疫特异性MAb分泌。酶联免疫吸附测定(ELISA)法最适于这一目的；但适用于大体积筛选的放射性免疫测定也是可接受的。可进行被设计用于连续地减少相当数量的无关或不太期望的培养基的多重筛选。可针对与已知B.t.杀虫毒素的交叉反应性筛选分泌与B.t.杀虫毒素反应的MAb的培养物。可使用可商购获得的测定鉴定优先与优选的B.t.杀虫毒素结合的MAb的同种型。优选的MAb是IgG类，更高优选的MAb是IgG₁和IgG_2a亚同种型。

可亚克隆数次分泌优选的MAb的杂交瘤培养物，以建立单克隆性和稳定性。用于亚克隆真核、非贴壁细胞培养物的熟知方法包括有限稀释、软琼脂糖和荧光激活细胞分选技术。在每次亚克隆后，优选针对抗体分泌和亚型再次测定所得培养物，以确保建立了稳定分泌优选MAb的培养物。

抗B.t.毒素抗体可用于检测本发明要求保护的B.t.杀虫毒素及其变体或片段的各种方法中。熟知用报告基团标记的抗体可用于鉴定各种环境中抗原的存在。用放射性同位素标记的抗体已在放射性免疫测定中用了数十年，其以高精度和灵敏度鉴定各种生物流体中抗原的存在。最近，酶标记的抗体已在ELISA测定中用作放射标记的抗体的替代手段。此外，可将与本发明的B.t.杀虫毒素免疫反应的抗体结合至固定物质，诸如聚苯乙烯孔或颗粒，并用于确定测试样品中是否存在B.t.毒素的免疫测定中。

使用探针检测.用于鉴定本发明的多肽和基因的又一方法是通过使用寡核苷酸探针。这些探针是可检测的核苷酸序列。这些序列可以依靠恰当的放射性标记而使得能够被检测，或者如美国专利No.6268132中所述的制作成固有荧光性的。如本领域所熟知，如果探针分子与核酸样品通过在这两个分子间形成强碱基配对键而杂交，则可以合理地假定该探针和样品具有相当的序列同源性。优选地，通过本领域熟知的技术在严格条件下进行杂交，例如如Keller和Manak(1993)所描述的那样。探针检测提供了以已知的方式确定杂交是否已经发生的手段。此种探针分析提供了鉴定本发明的编码毒素的基因的快速方法。作为根据本发明的探针使用的核苷酸区段可以使用DNA合成仪和标准方法进行合成。这些核苷酸序列还可以用作PCR引物，以扩增本发明的基因。

杂交.如分子生物学领域的技术人员熟知的那样，两个核酸分子的相似性可通过它们的杂交倾向来表征。如本文所使用的术语“严格条件”或“严格杂交条件”旨在指代这样的条件，在这样的条件下探针将以比其他序列更高程度(例如，比背景高至少2倍)可检测地与其靶序列杂交(退火)。严格条件是序列依赖性的并且在不同的环境中将是不同的。通过控制杂交和/或洗涤条件的严格程度，可鉴定与探针100％互补的靶序列(同源探测)。或者，可调节严格条件，以允许序列中的一些错配，从而检测较低程度的相似性(异源探测)。通常，探针少于约1000个核苷酸长，优选少于500个核苷酸长。

通常，严格条件将是以下这些：其中在pH 7.0至pH 8.3下盐浓度小于约1.5M Na离子，一般为约0.01至1.0M Na离子浓度(或其他盐)，并且温度对于短探针(例如，10至50个核苷酸)而言至少约30℃，而对于长探针(例如，超过50个核苷酸)而言至少约60℃。还可以通过加入去稳定剂诸如甲酰胺来实现严格条件。示例性的低严格条件包括在30％至35％甲酰胺、1M NaCl、1％SDS(十二烷基硫酸钠)的缓冲溶液在37℃杂交，并在50℃至55℃在1X至2XSSC(20X SSC＝3.0M NaCl/0.3M柠檬酸三钠)中洗涤。示例性的中等严格条件包括在40％至45％甲酰胺、1.0M NaCl、1％SDS中在37℃杂交，并在55℃至60℃在0.5X至1X SSC中洗涤。示例性的高严格条件包括在50％甲酰胺、1M NaCl、1％SDS中在37℃杂交，并在60℃至65℃在0.1X SSC中洗涤。任选地，洗涤缓冲液可包含约0.1％至约1％的SDS。杂交的持续时间一般少于约24小时，通常为约4至约12小时。

特异性通常是杂交后洗涤的函数，关键因素是最终洗涤溶液的离子强度和温度。对于DNA/DNA杂交，热解链温度(T_m)是50％的互补靶序列与完美匹配的探针杂交的温度(在确定的离子浓度和pH下)。每1％的错配，则T_m降低约1℃；因此，可调节T_m、杂交条件和/或洗涤条件以促进具有期望同一性的序列的退火。例如，若寻找具有＞90％同一性的序列，则T_m可降低10℃。通常，严格条件选择为比特定序列及其互补序列在确定离子强度和pH下的T_m低约5℃。然而，高严格条件可利用在比T_m低1℃、2℃、3℃或4℃下的杂交和/或洗涤；中等严格条件可利用在比T_m低6℃、7℃、8℃、9℃或10℃下的杂交和/或洗涤；并且低严格条件可利用在比T_m低11℃、12℃、13℃、14℃、15℃或20℃下的杂交和/或洗涤。

T_m(单位为℃)可实验确定或可通过计算估计。对于DNA-DNA杂交，可从Meinkoth和Wahl(1984)等式估计T_m：

T_m(℃)＝81.5℃+16.6(log M)+0.41(GC％)-0.61(甲酰胺％)-500/L；

其中M是单价阳离子的摩尔浓度，GC％是DNA中鸟嘌呤和胸腺嘧啶核苷酸的百分比，甲酰胺％是杂交溶液中甲酰胺的百分比(w/v)，并且L是杂交的碱基对长度。或者，T_m由以下公式描述(Beltz等人，1983)。

T_m(℃)＝81.5℃+16.6(log[Na+])+0.41(GC％)-0.61(甲酰胺％)-600/L

其中[Na+]是钠离子的摩尔浓度，GC％是DNA中鸟嘌呤和胸腺嘧啶核苷酸的百分比，甲酰胺％是杂交溶液中甲酰胺的百分比(w:v)，并且L是杂交的碱基对长度

使用上述的等式、杂交和洗涤组合物，以及期望的T_m，本领域的一般技术人员将理解，杂交和/或洗涤溶液的严格度的变化均已被固有地规定。如果期望的错配程度导致低于45℃(水溶液)或32℃(甲酰胺溶液)的T_m，优选增加SSC浓度，使得可使用更高的温度。核酸杂交的广泛指引可在Tijssen(1993)和Ausubel等人(1995)中找到。还参见Sambrook等人(1989)。

可通过标准方法(Sambrook等人，同上)进行使用放射标记的基因特异性探针，在DNA印迹上对固定的DNA的杂交。用于标记多核苷酸探针的放射性同位素可包括32P、33P、14C或3H。将放射性同位素掺入多核苷酸探针分子可通过分子生物学领域的技术人员熟知的若干种方法的任一种完成。(参见，例如Sambrook等人，同上)。一般地，可以在允许检测与所要求保护的杀虫蛋白编码基因具有同源性的靶序列的严格条件下进行杂交和随后的洗涤。对于双链DNA基因探针，可在6X SSPE、5X Denhardt溶液、0.1％SDS、0.1mg/mL变性DNA中，在比DNA杂交体的T_m低20℃至25℃下进行杂交过夜(20X SSPE是3M NaCl、0.2M NaHPO₄和0.02M EDTA(乙二胺四乙酸钠盐)；100X Denhardt溶液是20gm/L聚乙烯吡咯烷酮、20gm/LFicoll 400型和20gm/L牛血清白蛋白(级分V))。

洗涤通常如下进行：

室温下在1X SSPE，0.1％SDS中15分钟两次(低严格洗涤)。

在0.2X SSPE，0.1％SDS中在-20℃的T_m洗涤一次15分钟(中等严格洗涤)。

对于寡核苷酸探针，在6X SSPE、5X Denhardt溶液、0.1％SDS、0.1mg/ml变性DNA中在低于杂交体的T_m 10℃-20℃下进行杂交过夜。通过以下公式确定寡核苷酸探针的T_m(Suggs等人，1981)。

T_m(℃)＝2(T/A碱基对数目)+4(G/C碱基对数目)

洗涤通常如下进行：

室温下1X SSPE，0.1％SDS中15分钟两次(低严格洗涤)。

在1X SSPE，0.1％SDS中杂交温度下洗涤一次15分钟(中等严格洗涤)。

可通过除放射性标记之外的方法使用于杂交的探针分子及探针与靶分子之间形成的杂交分子可检测。此类替代的方法包含在本发明的范围之内。

本文通过使用术语“遗传材料”，其意思是包括所有的基因、核酸、DNA和RNA。术语“dsRNA”指双链RNA。为命名多核苷酸、DNA、RNA、寡核苷酸和引物的核苷酸残基，以及命名蛋白质的氨基酸残基，在本文件中使用了标准的IUPAC缩写。核酸序列以标准的5’至3’方向呈现，以及蛋白质序列以标准的氨基(N)端至羧基(C)端方向呈现。

除非特别指明或暗示，如本文所用的术语“一个”、“一种”和“该/所述”表示“至少一个/种”。

除非另外指明，所有百分比均以重量计，所有溶剂混合物比例均以体积计。所有温度均以摄氏度计。

本文提到或引用的所有专利、专利申请、临时申请和出版物都完整引入本文，以它们没有与本说明书的明确教导不一致为限。

应当理解，本文描述的这些实施例和实施方案应该解释为仅用于阐明的目的，并且在其教导下的各种修饰和改变对于本领域技术人员而言是容易想到的，并且包括在本申请的精神和范围之内，并且包括在所附权利要求的范围之内。这些实施例不应被理解为是限制性的。

实施例1

具有转运肽的DIG-177(野生型Cry6Aa)

到玉米中的转化

设计DIG-177B.t.杀虫多肽的编码序列的植物优化版本。植物分子生物学领域的技术人员将理解可以设计多个DNA序列以编码单个氨基酸序列。增加所关注的蛋白质的编码区表达的常用手段是以这样的方式调整编码区，使得编码区的密码子组成类似于基因注定表达的宿主的总体密码子组成。关于合成基因的设计和产生的指导可见于例如WO1997013402、美国专利No.6,166,302和美国专利No.5,380,831。

设计并合成具有玉蜀黍密码子偏倚的DNA序列，以在转基因单子叶植物中产生DIG-177杀虫蛋白(SEQ ID NO:2)。根据从GenBank(www.ncbi.nlm.nih.gov)中保藏的序列获得的数百种蛋白质编码序列计算玉蜀黍(Zea mays L.)的密码子使用表。在省略少于该氨基酸总密码子使用的约10％使用的任何同义密码子后计算重新调节的玉蜀黍密码子组。

为了得到编码DIG-177蛋白质的玉蜀黍密码子优化的DNA序列，DIG-177杀虫蛋白质或其杀虫片段的变体或嵌合体是对编码杀虫蛋白的实验确定的(天然的)DIG-177DNA序列(SEQ ID NO:1)进行密码子取代的受体，使得所得DNA序列具有玉蜀黍优化的密码子偏倚表的全部密码子组成。进一步改进序列以消除不期望的限制酶识别位点、潜在的植物内含子剪接位点、A/T或C/G残基的长期运行以及可能干扰mRNA稳定性、植物细胞中编码区的转录或翻译的其他基序。进行其他变化以引入期望的限制酶识别位点并消除较长的内部开放阅读框(除+1外的框)。这些改变都是在保留玉蜀黍偏倚的、重新调整的密码子组成的限制下完成的。编码DIG-177多肽的玉蜀黍优化的DNA序列被公开为SEQ ID NO:3。

构建体设计和质粒列表.所有构建体都是针对标准核转化、核转录和细胞溶质翻译而设计的。使用玉蜀黍泛素启动子和玉蜀黍过氧化物酶5 3'非翻译区来调节DIG-177编码区。除草剂选择由甘蔗杆状病毒启动子以及玉米条纹病毒前导序列和玉米醇脱氢酶内含子的控制下的AAD-1基因(Wright等人，2010)提供；3'非翻译区来自玉米脂肪酶基因。测试了七个不同的亚细胞隔室的DIG-177积累。表2显示了所用转运肽的汇总。表2

质粒、编码区和转运肽除了质外体外，对每个隔室测试多个转运肽。针对氨基末端转运肽去除天然DIG-177起始甲硫氨酸密码子。调整转运肽以反映与DIG-177编码区相似的密码子偏倚。

表2

质粒、编码区和转运肽的汇总

(N表示氨基末端位置；C表示羧基末端位置)

T₀和T₁实验设计每个构建体二十五个转基因事件的目标设定为有个为低拷贝。通过LC/MS/MS分析所有事件的插入拷贝数和DIG-177叶片积累。将低拷贝事件留下并丢弃未表达者，并将表达者接着用于T₁种子生产。这确保了在T₀生物测定中可能丢失的部分有效事件的恢复。多拷贝表达事件在T₀阶段进行生物测定；通过的那些被保存用于T₁种子生产和进一步分析。将T₁种子种植，分析植物的叶和根组织中的蛋白质(LC/MS/MS和蛋白质印迹)，在生物测定中测试到来了五个姊妹株。

农杆菌培养启动。含有DIG-177植物转化构建体(表2)的根癌农杆菌(Agrobacterium tumefaciens)菌株从DAS重组培养物保藏中心获得。将培养物从甘油贮存液中划线接种到农杆菌(Agrobacterium)基本培养基上，并在20℃黑暗中温育3天。然后将农杆菌属培养物划线接种到YEP培养基平板上并在20℃下在黑暗中温育1天。

实验当天，以适合于实验中的构建体数目的体积制备接种培养基和乙酰丁香酮的混合物。将溶于100％二甲亚砜的1M乙酰丁香酮储备溶液以适当的体积加入含有接种培养基的烧瓶，以制成终浓度为200μM的乙酰丁香酮。

对于每个构建体，将1-2个接种环的来自YEP平板的农杆菌属悬浮于50ml无菌一次性离心管内的15ml接种培养基/乙酰丁香酮混合物中，并且在分光亮度计中测定溶液在600nm下的光学密度(O.D.₆₀₀)。然后使用另外的接种培养基/乙酰丁香酮混合物将悬浮液稀释至0.25-0.35O.D.₆₀₀。然后在使用前将农杆菌属悬浮液管水平放置于设定为约75rpm、室温下的平台摇床上1至4小时。

穗消毒和胚分离。来自玉蜀黍品种B104的穗在温室设施中生产并在授粉后10-12天收获。实验当天，将收获的穗脱壳，并通过浸没在市售漂白剂(Ultra杀菌漂白剂，6.15％次氯酸钠)和两滴肥皂的20％溶液中并振摇20分钟来进行表面消毒，随后置于层流罩内在无菌去离子水中漂洗三次。从每颗穗无菌切除未成熟的合子胚(1.8-2.2mm长)，并分配到含2.0ml农杆菌属悬液的一个或多个微量离心管中，该悬液中已加入2μl的10％S233表面活性剂。

农杆菌共培养。在完成胚胎分离活动后，封闭胚胎管并放置在摇床上5分钟。然后将管的内容物倒到共培养培养基平板上，并用无菌一次性移液管移除液体农杆菌悬液。将含有胚胎的共培养平板置于盖半开的层流罩背面30分钟；此后，使用显微镜将胚胎定向为胚子叶朝上。然后将带有胚胎的共培养平板返回到盖半开的层流罩的背面另外15分钟。然后盖上平板，用3M Micropore^TM胶带密封，然后放置在具有大约60μmol m^-2s^-1光合有效辐射(PAR)的24小时/天光照的25℃培养箱中。

选择愈伤组织和再生转基因事件。在共培养期后，将胚胎转移到静息培养基中。将不超过36个胚移到每个平板上。将平板在27℃下，使用约50μmol m^-2s^-1PAR的24小时/天光照温育7-10天。然后将愈伤组织胚胎转移到选择I培养基上。将平板在27℃下使用约50μmolm^-2s^-1PAR的24小时/天的光照温育7天。然后将愈伤组织胚胎转移至选择II培养基。将平板在27℃下使用约50μmol m^-2s^-1PAR的24小时/天的光照温育14天。

在此阶段，抗性愈伤组织被移至预再生培养基。将平板在27℃下使用约50μmol m^{- 2}s^-1PAR的24小时/天的光照温育7天。然后将再生中的愈伤组织转移到Phytatrays^TM中的再生培养基上，并在28℃下以每天16小时光照/8小时黑暗(大约150μmol m^-2s^-1PAR)温育7-14天，或直到发出芽为止。然后分离具有初生根的小芽，并转移到强健培养基上。将约6cm或更高的生根小植株移植到土壤中并移至生长室以炼苗(hardening off)。

从植物组织进行基因组DNA分离以用于PCR将等同于2个叶冲孔块(punch)的叶撕裂物组织样品收集在96孔的收集平板(凯杰公司(Qiagen)，德国希尔登)中。用配备一颗不锈钢珠的Klecko^TM组织粉碎器(Garcia制造公司(Garcia Manufacturing)，加利福尼亚州维塞利亚)在Biosprint96AP1裂解缓冲液中进行组织破碎。在组织浸渍之后，以高通量形式分离基因组DNA。在建立qPCR反应之前，将基因组DNA稀释成2:3的DNA/H₂O，以实现合适的Cp评分。

水解探针测定.通过实时PCR，借助水解探针测定来执行转基因检测。使用探针设计软件2.0在Dow AgroSciences处针对所关注的基因(GOI)Cry6Aa(IRDIG522.28)设计测定。使用玉蜀黍TIP41样作为参考，将RNA检测测定作为单一反应进行。为了扩增，制备了在10μL体积的双重反应物中的1x最终浓度的探针母液混合物(Roche Applied Science)，其含有每种引物各0.4μM，以及每种探针各0.2μM。采用荧光采集在60℃延伸40秒进行两步扩增反应。使用拟合点算法(软件发行版本1.5)和绝对定量模块(基于ΔΔCt方法)的Cp得分(荧光信号与背景阈值相交的点)用于执行实时PCR数据分析。

T₀温室.将承担选定事件的植物移植到5加仑的盆中。在穗丝露出之前将新果穗袋置于上以防止杂散花粉的交叉污染。然后覆盖第二个芽，并用于授粉。在授粉前一天将穗丝剪下来以提供均匀的刷子来接受花粉。来自近交B104的花粉用于所有授粉。在可能时执行互交。在授粉后21天将穗剥皮以增强干倒，然后在授粉后42天完成收获(从植株去除穗)。将穗置于干燥器中1周，然后进行种子处理(脱壳、计数、包装)。

用于蛋白质印迹和LC/MS/MS分析的叶取样.在生物测定收集的当天内，在V-3至V-5阶段对植物进行取样。将两个6mm直径的叶样品贮藏在-80℃的96孔集束式管架中，直到分析当天。将两个Daisy^TM钢BB和300μl提取缓冲液(含有0.05％吐温20和5μl/ml西格玛蛋白酶抑制剂的PBS溶液)加入到每个管中。在最大设定下将样品在Kelco珠磨机中研磨3分钟。将样品以3,000x g离心5分钟；将100μl上清液转移到空样品管中。将另外100μl的提取缓冲液加入到植物样品中，珠磨另外3分钟，离心，并将100μl的此提取物与第一个100μl混合。将合并的上清液在提取的同一天混合并分析。

用于蛋白质印迹和LC/MS/MS分析的根取样收集完整的根系，用水洗涤，并在V-3至V-5阶段将所有T₁植株保存在-80℃下。每个植株在单独的塑料袋中保留作为单独的样品。每个根系在工业食品加工机中用干冰研磨，干冰在-20℃升华，从而保持样品冷冻。然后将经研磨、升华的冷冻样品冻干2周。称重冻干根系的多个部分并放置在96孔格式的单独集束式管中。如上所述制备样品以用于叶取样。

通过LC/MS/MS进行DIG-177定量.在25M碳酸氢铵+0.05％Tween 20中提取每个植株的叶样品。样品通过珠磨提取两次，然后合并。对于根样品，将经研磨和冻干的根组织称重到小瓶中并如上所述进行提取。样品在95℃下用6mM DTT变性20分钟。样品冷却后，向每个样品中加入1μg胰蛋白酶，并将样品在37℃下消化16小时。消化后，将样品用1％甲酸处理，在4℃温育30分钟，并以4,000rpm离心10分钟。通过LC/MS/MS分析消化的样品。在消化的B104基质(如上所述制备)中制备DIG-177肽标准品的标准曲线。标准曲线从0.488-31.25ng/mL是线性的。使用Acquity UPLC BEH130C18、1.7μm的2.1×50mm柱，在WatersAcquity二元泵LC和Sciex QTRAP 6500上分析样品。每个样品以20μl注入，并使用快速梯度洗脱(95％-45％的A达1分钟，45-10％的A达1分钟，保持10％达1分钟，10-95％达1分钟：缓冲液A：水+0.1％甲酸，缓冲液B：乙腈+0.1％甲酸)。跟踪每个样品中的两种肽(m/z 694.820和m/z 586.778)。定量基于最灵敏的转变，m/z 694.820至_m/_z 912.417。

蛋白质印记.常规电泳和印迹(Galla_gher等人，2008)方法与Invitrogen^TM装置和基本试剂一起使用。兔抗Cry6a抗体是用于检测叶和根中的Cry6a的一抗。用荧光检测系统和抗生物素蛋白-碱性磷酸酶检测系统检测所有蛋白质。

T₀和T₁玉米根生物测定在来自Crop Characteristics公司(明尼苏达州法明顿)的土壤中接收非滞育的玉米根叶甲(Diabrotica virgifera)(西方玉米根虫，WCR)卵并在28℃和60％RH下温育10天。在温育开始后第10天(大约预计孵化前一天)，制备得到卵用于生物测定。从卵上冲洗掉土壤。将卵以每0.5ml约100-200个卵的浓度悬浮在0.15％的琼脂水溶液中，并放入孵化平板中。对孵化平板上的活卵数目进行计数以确定悬浮液中卵的近似浓度，并因此计算实现所需侵染而需要添加到生物测定设施中的悬液体积。将平板保持在黑暗盒中的28℃培养箱中。

用于WCR生物测定的种植和制备将来自实验室的转化植物(T₀)移到环境中，将它们在该环境中保持，直到它们达到V3-V4生长阶段。将具有良好根系的V3-V4健康植物移至温室。将弱、小、或根发育差的植株留在环境中，直到健康到足以进行生物测定。在温室中一到两天后，将植物移植到装有Metro-Mix土壤的根整形容器(root trainer)中。

在预期获得用于生物测定的第一T₀植株之前至少两周开始，每周将对照植物的种子直接种植在根整形容器盆中的Metro Mix土壤中。移植T0植株约4天后，将植株与处于同一生长阶段的每种类型的五个对照植株一起用WCR卵侵染。

阴性对照植株如下：B104近交系，7SH382Herculex RW(HX-RW)近交系和101556(一种黄色荧光蛋白转基因的)。阳性对照是玉米系7SH382RW转基因Herculex RW(HX-RW)。

植株被大约200个卵/植株侵染。假设孵化率为约50％，由此预计每个植株将有大约100个幼虫孵化并以根为食。在整个生物测定过程中仔细监测浇水，以确保每个盆保持湿润。

侵染两周后，从对照植物开始，对每个植株进行根伤害评分。小心地将每个植株从根整形容器盆中取出，保持根部团块完整。从根部刮去顶部一至两英寸的土壤，并将所得到的根部暴露区域用水洗涤以便清楚地观察植物基部(节生根顶部)的进食损伤。出于温室玉米幼苗根损伤评级需要，使用0.0至1.0根损害评定量表来对进食损伤进行评分，所述评定量表来源于公布的田间玉米根部损伤评定量表0.0至3.0(J.D.Oleson等人，2005)。阴性对照(RW近交系、B104和101556)预计具有高度损伤(约0.75-1.0根系评级)，而阳性对照(转基因Herculex^TM RW(HX-RW))预计具有低损伤(0.0到0.25根评级)。在完成对照植株的评估之后，采用合格/不合格方法(0-1.00评分)并尽可能少干扰根球来对T₀植株评分。立即将合格的植株移植到具有promix-50-50土壤的5加仑盆中。在对所有植株评分后，将合格的植株用1茶匙FORCE(人工受精用七氟菊酯)的土壤施用处理，并在整个发育过程中适当护理。

T₁温室.将选出的植株移栽到5加仑的盆中。在穗丝露出之前将新果穗袋置于上以防止杂散花粉的交叉污染。然后覆盖第二个芽，并用于授粉。在授粉前一天将穗丝剪下来以提供均匀的刷子来接受花粉。使用来自植株雄穗的花粉来授粉同一植株上的穗。在可能时执行自花授粉。如果不能进行自花授粉，则将B104花粉带到转基因穗上。在授粉后21天将穗剥皮以增强干倒，然后在授粉后42天完成收获(从植株去除穗)。将穗置于干燥器中1周，然后进行种子处理(脱壳、计数、包装在预先印制的信封中)。

来自T₀事件的蛋白积累结果.获得8至28个表达DIG-177的低拷贝事件，整体转化频率为9％。通过LC/MS/MS分析来自十五个构建体中的每一个的通过分子分析筛选的所有再生事件的DIG-177叶蛋白积累。平均蛋白质积累显示于表3中。平均T₀叶蛋白质积累水平在0ng/cm²至98.9ng/cm²范围内，其中构建体pDAB117255靶向于前液泡(prevacuole)，具有最低的平均积累量，而构建体pDAB117261靶向叶绿体，具有最高的平均积累。总体而言，从将DIG-177蛋白靶向叶绿体的两种构建体pDAB117260和pDAB11261分别看到最高的平均蛋白质积累水平。这两种构建体的蛋白质积累高于先前的细胞溶胶实验中观察到的平均为23.6ng/cm²的蛋白质积累。相反，靶向液泡、前液泡和质外体的构建体导致最低的叶蛋白质积累。

表3

通过LC/MS/MS测定的平均DIG-177蛋白质叶积累。

通过蛋白质印迹分析来自十五个构建体中的每一个的选择T₀事件的叶组织。通过使用转运肽将蛋白质引导到特定的隔室。因此前体蛋白比54.2kDa的全长野生型DIG-177蛋白具有更大的分子量。取决于隔室，转运肽(TraP)可能或可能不会在易位后被切割，从而产生成熟形式的蛋白质。表4显示了蛋白质的前体和成熟形式的预测分子量。

表4

关键为在本研究中使用的转运肽修饰的DIG-177蛋白的质粒、蛋白名称、靶隔室和分子量。

非转基因阴性对照样品未显示与DIG-177抗血清反应的任何背景条带。在转基因样品中，在许多样品中检测到了与被去除的转运肽一致的明显成熟蛋白质(与DIG-177标准品共迁移)的独特条带。在一些情况下，还检测到比成熟蛋白质更大的条带，其与检测到前体蛋白质(转运肽未被去除)一致。所有构建体都显示出根据明显成熟条带54kDa至约40kDa的DIG-177材料的成片条带，其与蛋白质的部分降解一致。

T₀事件的生物测定结果.用在V3-V4发育阶段的WCR幼虫刺激来自代表15种不同背景的事件的根系达两周。然后使用如上所述的合格/不合格评定系统对针对对根系的损伤而对样品进行评分。共检测了228个事件，其中15个事件被针对T₁生物测定评分为“合格”并保存。

T₁事件的分析。分析T₁事件中叶和根组织中转基因蛋白的产生和积累。从T₀到T₁代的十五个背景中的每一个的平均叶蛋白质积累增加并且范围为从11.6到731.5ng/cm²(上表3)。在所有背景下，T₁根积累显着低于T₁叶片积累(表5)。

表5

通过LC/MS/MS测定的叶和根组织的事件平均DIG-177T₁蛋白质

积累

通过蛋白质印迹分析来自每个T₁事件的叶和根组织的蛋白质稳定性。结果与在T₀中看到的那些结果类似；明显的成熟蛋白质(与DIG-177标准品共迁移)的不同条带与被去除的转运肽一致。此外，还检测到比成熟蛋白质更大的条带，其与前体蛋白质一致。最终所有构建体都产生约54kDa至约40kDa的DIG-177材料的成片条带，其与蛋白质的部分降解一致。

T₁事件的生物测定结果.用在V3-V4发育阶段的WCR幼虫刺激来自代表15种不同背景的事件的五个植株达两周。样品如前述针对根系损伤进行分级；根评级≤0.5的植株被认为合格。所有测试的事件均未通过此生物测定；在根保护方面，没有任何事件与阴性对照显著不同。因此，使用转运肽定位至各个单独的亚细胞隔室并不能充分保护DIG-177免于蛋白酶降解。

蛋白质印记分析.图6示出了T₁玉蜀黍叶表达实验的结果。蛋白质印记分析显示在阴性对照；B104(泳道2)和pDAB115782(泳道3)或在含有YFP和PAT的转化对照(泳道12)中没有Cry6Aa蛋白表达。六条最右边的泳道是DIG-1000的蛋白质标准品(47.3kDa)。

pDAB117261(DIG-177)泳道3、8、11和14含有T₁样品作为对照以与使用DIG1000的处理进行比较。结果与在T₀中看到的那些结果类似；明显的成熟蛋白质(与DIG-177标准品共迁移)的不同条带与被去除的转运肽一致。此外，还检测到比成熟蛋白质更大的条带，其与前体蛋白质一致。这种对照构建体产生约54kDa至约40kDa的DIG-177材料的成片条带，其与蛋白质的部分降解一致。

靶向过氧化物酶体的pDAB126937(DIG-1000)的结果位于图6的泳道5和10中。存在以对应于DIG 1000标准品的正确大小产生的一个主要条带。泳道10含有略低于使用DIG177产生的40kDa成片条带的一个明显较小的条带，指示一个特定的加工位点。

靶向线粒体pDAB126938的DIG-1000位于图6的泳道4、6、9和13中。此构建体产生两个条带，一个条带具有正确大小，而另一个条带具有略高的大小，可能代表前体蛋白质。这些数据与表达可用于西方玉米根虫控制的植物细胞稳定的杀虫蛋白的DIG-1000编码序列一致。

实施例2

DIG-177二硫键的鉴定

重组蛋白质表达质粒.将DIG-177和DIG-177变体编码序列克隆到单独的假单胞菌(Pseudomonas)表达载体pDOW1169中。然后将表达载体转化到荧光假单胞菌(Pseudomonasfluorescens)中，并对所得转化菌落进行鉴定和贮藏。

重组蛋白质表达.将毫升甘油贮存液接种到2.5L Ultra Yield烧瓶(汤姆森仪器公司(Thomson Instrument Company)，加利福尼亚州欧申赛德)中的500mL Ultra^TM生产培养基中，并在30℃温育24小时，用1英寸长的摇瓶以225rpm的转速振荡。用150μL的1M IPTG诱导表达。将细胞在25℃温育24小时，此时取0.5mL样品并从摇床中取出烧瓶。通过在JA-20转子(贝克曼库尔特公司(Beckman Coulter)，加利福尼亚州布雷亚)中以19,000rpm离心10分钟或在FIBERLite^TM F12-6X500LEX转子(赛默飞世尔科技公司(Thermo Scientific))中以11,409rpm离心15分钟来收获培养物；将细胞体贮藏在-80℃下。

重组蛋白质富集.将DIG-177和下面描述的变体蛋白质作为包涵体(IB)积累在荧光假单胞菌(P.fluorescens)中。将细胞体在4℃解冻。将细胞在裂解缓冲液(50mM Tris，pH7.5，200mM NaCl，10％甘油，20mM EDTA，0.5％Triton X-100，5mM苯甲脒，以及在使用之前加入的1mM DTT)中悬浮成10％w/v，并用匀化器混匀。将浆液以12000+psi通过微流体微流化器(Microfluidics Microfluidizer)(韦斯特伍德，美国马塞诸塞州)两次。将裂解物在4°以14,000x g离心40分钟。保留上清液。将包涵体沉淀在裂解缓冲液中重悬至10％w/v，并加入溶菌酶(L-7651；西格玛奥德里奇公司(Sigma-Aldrich)，密苏里州圣路易)至终浓度为400μg/mL。将悬浮液在室温(约20℃)下温育60分钟，并离心。将包涵体沉淀用裂解缓冲液洗涤一次并用删去Triton^TM X-100的裂解缓冲液洗涤第二次。最后，将包涵体沉淀物在10mMEDTA溶液中重悬至30％w/v并在-80℃以2mL等分试样贮存。通过SDS-PAGE(NuPAGE Bis-Tris凝胶，协定公布No.MAN0007894，修订版A.0，英杰生命技术公司(Invitrogen Lifetechnologies)，马塞诸塞州沃尔瑟姆)分析样品纯度。

包涵体增溶.将储存在-80℃的IB糊在4℃下解冻过夜，然后在4℃以23,000×g离心25分钟，去除上清液。将IB颗粒在100mM CAPS(pH 11)中悬浮至20(w/v)中，并在室温下增溶2小时。如上所述离心样品并收集上清液。通过PD-10脱盐柱将缓冲液交换到10mM CAPS(pH 10)中。按照“Quick Start TM Bradford Protein Assay-Bio-Rad”(伯乐公司(Bio-Rad)，加州赫拉克勒斯)协定，通过Bradford蛋白质测定对蛋白质进行定量。标准品是BSA(来自Pierce公司，纽约州格兰德岛的牛血清白蛋白，白蛋白标准品)。

SDS-PAGE.按照来自英杰生命技术公司(协定公布No.MAN0007894，修改版A.0，Ref.2)的协NuPAGE Bis-Tris凝胶进行SDS-PAGE。简而言之，将样品与变性样品缓冲液(英杰公司)加5mM DTT(用于减少PAGE)或减去DTT(用于非还原PAGE)混合并在95℃加热5分钟。将样品加载到Invitrogen Bis/Tris凝胶上。用MOPS或MES跑样缓冲液将凝胶在200V下跑样约43分钟。将凝胶用考马斯蓝染色液(伯乐公司)染色30分钟，并用脱色液(7％乙酸和5％甲醇水溶液)脱色，直至背景色被清除。

完整的分子量分析/带电状态分布.使用加热至50℃的Michrom脱盐阱在Agilent1200HPLC/MSD TOF 1969A系统上进行完整质量分析。在10mM CAPS,pH 11缓冲液中将每种样品稀释至0.2μg/μL的浓度。还将样品在用50mM TCEP((2-羧乙基)膦盐酸盐)还原10分钟后进行分析将约1μg蛋白质注入柱上。使用梯度(10％缓冲液D达1分钟，10-60％缓冲液D达2分钟，60-98％缓冲液D达2分钟，10％缓冲液D达1分钟)洗脱样品，其中缓冲液A为0.1％的甲酸水溶液，而缓冲液D是70％异丙醇、20％乙腈、10％水+0.1％甲酸。使用Mass Hunter定性分析软件和最大熵解卷积算法计算质量。

对DIG-177在还原和非还原条件下的质谱分析证实蛋白质中存在两个二硫键。DIG-177具有位于位置88、162、402、404和451处的五个半胱氨酸残基。为了鉴定促成二硫键的残基，在选定的半胱氨酸处进行突变并表达重组蛋白质。在存在或不存在还原剂的情况下分析变体蛋白质的完整分子量。在这些条件下，当用TCEP还原时，蛋白质中存在的每个二硫键预期会导致m/z增加2。在还原野生型蛋白质(DIG-177)后，观察到m/z增加3.8，这指示蛋白质中存在两个二硫键。i DIG-614(SEQ ID NO:35)、615(SEQ ID NO:37)、618(SEQ IDNO:43)、619(SEQ ID NO:45)、983(SEQ ID NO:47)和984(SEQ ID NO:49)在还原后各自表现出约+2的质量变化，这代表单个二硫键。预计这些变体中的每一个都会导致半胱氨酸残基88与451之间的二硫键消失。基于这些数据，存在涉及半胱氨酸残基162、402或404中的两个的附加二硫键。与DIG-177一样，预期破坏参与Cys162的二硫键的突变体DIG-616(SEQ IDNO:39)和617(SEQ ID NO:41)在还原后显示约+4的变化，这指示存在两个二硫键。由于在半胱氨酸88和451之间发现了这些二硫键中的一个，推断第二个二硫键位于半胱氨酸残基402与404之间。

表6

加减使用TCEP进行还原的半胱氨酸突变体的完整MW.

实施例3

经胰蛋白酶处理的DIG-177的表征

概括地说，由一个大片段(LF；SEQ ID NO:50的氨基酸12-390)和两个小片段中的任一个(氨基酸444-475(SEQ ID NO:51)或451-475(SEQ ID NO:52))组成的经胰蛋白酶处理的DIG-177(胰蛋白酶核心，TC)称为羧基末端肽(CTP)。发现LF和CTP通过cys88与cys451之间的二硫键连接。在非还原条件下，经胰蛋白酶处理的样品具有杀虫活性；而在还原条件下，片段解离(LF和CTP)并且活性降低。LF和CTP都没有显示出测试水平的杀虫活性。这些观察结果表明，附着于LF的CTP对于维持胰蛋白酶蛋白水解后的毒素活性是必需的。

对制备、鉴定和测试的胰蛋白酶处理样品的概述.制备四种不同的样品，鉴定并进行生物测定以确定哪种肽组合具有WCR杀虫活性(参见图1)。

样品1：经胰蛋白酶处理的DIG-177，存在CTP和与LF连接的二硫键。

样品2：经胰蛋白酶处理的DIG-177，存在CTP，该样品经消化后用还原剂处理；然后通过渗析去除还原剂。

样品3：胰蛋白酶处理的DIG-177，分离掉LFT

样品4：胰蛋白酶处理的DIG-177，分离掉CTP

胰蛋白酶消化.如实施例2中所述，DIG-177蛋白质在荧光假单胞菌(Pseudomonasfluorescens)中产生并被纯化。将重组蛋白在含有胰蛋白酶的(经TPCK处理的，西格玛奥德里奇公司，密苏里州圣路易)的消化缓冲液(100mM CAPS缓冲液，pH 10.5，5mM CaCl₂，5mMMgCl₂)中用胰蛋白酶处理至最终比率为15份蛋白质比1份胰蛋白酶(W/W)。在室温下使用温和的摇动搅拌反应物。在每个时间点(5分钟、2小时和17小时)取出等分试样进行分析，通过加入含有5mM DTT的十二烷基硫酸锂(LDS)样品缓冲液并在95℃加热5分钟来终止消化。

使用AKTA蛋白质纯化器(安发玛西亚生物技术公司(Amersham PharmaciaBiotech)，新泽西州皮斯卡塔韦)通过离子交换色谱法(IEC)纯化经胰蛋白酶处理的蛋白质。用10mM CAPS(pH10)以1:1稀释样品，并通过具有0.22μM膜的真空驱动过滤器过滤。将样品以3mL/min的流速注射到预平衡的HiTrap^TM Q柱(通用电气公司(GE)，宾夕法尼亚州匹兹堡)上，用约25mL缓冲液A(50mM CAPS，pH 10)洗涤柱，将蛋白质用20％缓冲液B(缓冲液A+1MNaCl)洗脱，然后用30％、40％和50％缓冲液B洗脱。每次洗脱液约25毫升。基于紫外吸光度分别从各个洗脱液收集级分。通过SDS-PAGE分析这些级分(参见来自英杰生命技术公司，协定公布No.MAN0007894，修订版A.0的协定NuPAGE Bis-Tris凝胶)。

合并含有靶蛋白的级分并使用具有10kDa截留分子量(MWCO)膜(密理博公司(Millipore)，马塞诸塞州比勒利卡)的离心过滤装置浓缩至约3mL。通过渗析将样品缓冲液交换到10mM CAPS(pH 10)中。将浓缩的样品转移到Slide-A-Lyzer^TM渗析盒(赛默飞世尔科技公司)中。将盒置于4L的10mM CAPS(pH 10)中，并在4℃渗析过夜(约18小时)。收集通过SDS-PAGE(如实施例2中所述)评估为具有大于95％纯度的样品以分别用于WCR生物测定和/或鉴定。

按照制造商的协定“Quick Start^TM Bradford Protein Assay-Bio-Rad”(伯乐公司)，通过Bradford蛋白质测定法测量蛋白质浓度。使用BSA(牛血清白蛋白)作为标准品。

样品1：胰蛋白酶处理的DIG-177(LF加CTP)的制备和鉴定.将野生型DIG-177；SEQID NO:2)用胰蛋白酶消化16小时并在离子交换柱上跑柱以去除胰蛋白酶。在还原-变性条件下通过SDS-PAGE(如实施例2中所述)分析样品。未消化的全长物质以约54kDa迁移，在大致约108kDa处出现了少量二聚体。DIG-177可形成可检测量的高分子量聚集体。基于胰蛋白酶切割位点，经胰蛋白酶处理的样品在分级分离之前作为代表LF的约43kDa的单一条带，被预测为残基12-390。

N末端测序.LF的氨基末端通过使用基本Edman降解化学在Shimadzu蛋白质测序仪(型号PPSQ-33A)上进行的Edman降解N末端测序来确认。蛋白质样品在还原条件下用SDS-PAGE分离，然后通过液体转移将蛋白质印迹到PVDF膜上)。将靶蛋白条带切下并装载到玻璃反应腔室中。然后将腔室插入测序仪并进行Edman降解。每次运行20种PTH-氨基酸的标准混合物(岛津公司(Shimadzu)，日本京都)。来自每个Edman降解循环的氨基酸残基是基于其与标准品相比在C-18柱的保留时间确定的。顺序地重复此过程以获得该蛋白质的氨基末端序列。Edman降解确定LF的N末端从SEQ ID NO:2的残基12开始。

在没有还原的情况下进行蛋白完整分子量分析(如实施例2中所述)，产生了质量为46,844.19Da的优势物种和质量为45,995.25Da的次要物种(表7)。为46,844.19Da的去卷积完整MW平均值与针对包含二硫键键合到链444-475(CTP1)的12-390的多肽链的蛋白质预期的理论值(46,843.43Da)相匹配。MW平均值为45,995.25Da的次要物种与含有二硫键键合到链451-475(CTP2)的12-390的多肽链的蛋白质的理论值45,996.51Da相匹配。

表7

对在非还原条件下的样品1的完整分子量分析

在TCEP处理(还原条件)后测定样品的完整分子量；观察到了两种组分(表8)。42,729.71Da的主要组分匹配针对经胰蛋白酶处理的DIG-177LF(残基12-390，无CTP)的理论质量42,729.4。还观察到质量为46,844.14Da的次要组分，其匹配具有仍然附着的CTP1(不完全还原)的LF的质量。结果指示CTP1和CTP2通过用TCEP还原而大部分去除。

表8

对在还原条件下的样品1的完整分子量分析

样品2：胰蛋白酶处理的DIG-177(LF具有还原后的CTP)的制备和鉴定.如针对样品1所述在非还原条件下纯化胰蛋白酶处理的DIG-177。将DTT以5mM的终浓度加入至样品中，并在室温下温育10分钟。然后将样品用10mM CAPS(pH 10)渗析过夜以去除DTT而产生样品2。使用4-12％Bis/Tris凝胶和MOPS跑样缓冲液通过SDS-PAGE分析样品1和2以获得约40kDa的较好分辨率。在还原条件下，样品1和样品2(经DTT处理的)显示与LF(12-390)一致、在预期分子量约43kDa处的一个条带，凝胶底部的第二条带是(CTP1和CTP2在这些条件下不分解)已经通过二硫键还原释放的CTP。在非还原条件下，样品1跑样至约46kDa的预期分子量处，与连接有CTP的LF(残基12-390)一致。样品2(经DTT处理、去除的)显示两个较高分子量条带，一个在约43kDa处，与LF(残基12-390)一致；第二个为约46kDa条带，与LF加CTP一致。第三条带存在于凝胶底部，这与CTP对应。这些结果表明，在样品2中，大多数CTP不与大片段连接；样品的一部分看起来重新缔合。

没有TCEP处理的光谱显示测量的MW平均值为42,729.46Da、45,994.89Da和46,844.14Da的三个主峰。(表9)。42,729.46峰与LF(残基12-390)的理论质量(42,729.0Da)相匹配。45,994.89峰与LF加CTP2(残基12-390+451-475)的理论质量(45,995.0Da)相匹配。46,844.14组分与LF加CTP1(残基12–390+444-475)的理论MW平均值(46,843.72Da)相匹配。

表9

对在非还原条件下的样品2的完整分子量分析

使用50mM TCEP处理的光谱显示只有MW平均值为42,729.27的一种组分(表10)；在这个光谱窗口中没有捕获到CTP。这些结果证实DTT治疗释放了CTP，并且在去除DTT后，CTP的一部分与LF重新缔合。

表10

对在还原条件下的样品2的完整分子量分析

样品3：LF的制备和表征.为了分离DIG-177胰蛋白酶产生的LF和CTP，在还原条件(5mM DTT)下进行胰蛋白酶消化后的IEC纯化。通过非还原性SDS-PAGE分析样品。胰蛋白酶处理的非还原样品以约46kDa的预期分子量迁移，这与连接至CTP的LF(12-390)一致。起始材料样品(在制备过程中用DTT处理过的)迁移速度明显更快并且与CTP的损失一致。减去CTP的LF在级分3中获得。

为了验证级分3的组成，使样品经受完整的分子量分析(表11)。没有TCEP处理的光谱表现出MW平均值为42,729.76Da的优势物种和MW平均值为45,995.19Da的次要组分。42,729.76Da的主要组分与胰蛋白酶LF(残基12-390)的理论MW平均值(42,729.0Da)相匹配，这证实CTP被去除。45,995.19Da的次要组分与由LF(残基12-390)和CTP2(残基451-475)组成的第二物种的理论MW平均值45,995.0Da相匹配，这指示纯化期间的还原不完全。这些结果确认了大部分样品代表没有CTP的LF。

表11

对在还原条件下的样品3的完整分子量分析

样品4：C末端肽(CTP)的分离和鉴定。将在非还原条件下纯化的胰蛋白酶消化的DIG-177(见上文)用5mM DTT处理以释放C末端肽。将经处理的样品穿过具有10kDa MWCO(密理博公司)的离心过滤装置。C-末端肽存在于通过流中，而大片段保留在过滤单元中。收集流过和保留的样品，并通过渗析在10mM CAPS(pH 10)中交换缓冲液。

将C末端肽样品(样品4)通过MALDI-TOF/TOF MS进行分析以确定肽质量，并随后在LIFT模式下片段化以确认肽的序列。使用Bruker UltraFlextreme^TM质谱仪进行MALDI-TOFMS和MS/MS。用0.2％TFA(三氟乙酸)以1:1(v/v)稀释C末端肽样品并使用C-18Ziptip(密理博公司)脱盐。将肽用0.1％TFA中的60％乙腈(ACN)洗脱并与2,5-二羟基苯甲酸(DHB)基质(在ACN:H₂中为15mg/ml(50:50))混合。在MALDI样品平板上点出1μL混合物后，使用反射-阳性模式分析肽质量，并使用LIFT模式将肽片段化。将仪器用CM 2(校准混合物2，肽质量标准试剂盒，Applied Biosystems Sciex公司，加利福尼亚州福斯特市)校准。收集质谱并使用挠曲分析软件分析。使用BioTools软件(布鲁克公司(Bruker))和MASCOT搜索引擎(MatrixScience公司，马塞诸塞州波土顿)验证序列。MALDI MS显示了质量分别为4,114.894Da、3,266.38Da和2,154.933Da的三个主峰，以及若干个小峰(表12)。在4,114.89m/z处的峰与代表残基444-475(CTP1)的理论质量4,113.66相匹配；在3,266.38m/z处的峰与代表残基451-475(CTP2)的理论质量3,265.26相匹配。在2,154.933m/z处的峰不匹配来自C末端的任何肽并且其来源尚未确定。

表12

对在还原条件下的样品2的完整分子量分析

用MALDI MS/MS对两种肽m/z4,114.894Da和3,266.38Da进行测序。结果证实4,114.894峰具有与SEQ ID NO:51的残基444-475构成的CTP1序列匹配的序列NSNLEYKCPENNFMIYWYNNSDWYNNSDWYNN(加下划线的残基被鉴定和指定)。3,266.38峰具有与来自SEQ ID NO：52的残基#451–475的CTP2序列匹配的序列CPENNFMIYWYNNSDWYNNSDWYNN。

样品1至4的杀虫活性.非滞育WCR卵(Crop Characteristics公司，明尼苏达州法明顿市)在28℃下在土壤中温育10天。将这些卵用水从土壤中洗出，用10％甲醛表面灭菌并用无菌水冲洗三次。使这些卵孵化并用专有的WCR饮食饲喂。在24孔滴定板中进行叠加饮食生物测定，其中每个孔含有1.5mL人造WCR饮食。将测试样品以100μg/cm²剂量(80μL)(除非另有说明)施用到饮食表面上，并在室温下以层流干燥。将每孔的经处理的饮食表面用五只新生玉米根虫(D.virgifera)(24-48小时龄)侵染，并将测试昆虫封装在具有Breathe透气盖的生物测定平板中，将该平板密封并保持在受控的环境条件下(28℃，24小时人工黑暗期，60-80％相对湿度)。每个平行测定测试二十只昆虫。在温育五天后记录每次处理的活昆虫数量、死昆虫数量和合并的昆虫活体重量。记录死亡昆虫的数量和存活昆虫的重量。

计算每个处理的平均死亡率百分比和平均生长抑制。生长抑制(GI)如下计算：

GI＝[1–(TWIT/TNIT)/(TWIBC/TNIBC)]

其中TWIT是处理中昆虫的总重量，TNIT是处理中的昆虫总数，TWIBC是背景检查(缓冲液对照)中的昆虫总重量，并且TNIBC是背景检查(缓冲液对照)中的昆虫总数。阴性对照是20mM柠檬酸钠缓冲液(pH3.5),10mM CAPS缓冲液(pH 10)和350μg/cm²Cry1Fa。阳性对照是在柠檬酸钠缓冲液中的100μg/cm²Cry34/35Ab1和/或在CAPS缓冲液中的100μg/cm²DIG-177。以100μg/cm²(DIG-177的LC₅₀为约40μg/cm²)对来自经胰蛋白酶处理的DIG-177的样品1-4进行生物测定。生物测定结果汇总于表13中，Cry1Fa和两种缓冲液样品用作阴性对照。使用全长DIG-177作为阳性对照。

样品1，大片段+CTP，显示与全长阳性对照几乎等同的杀虫活性。样品2，释放掉CTP但存在于样品中的大片段，具有低但可测量的活性，这表明用DTT处理然后渗析显著降低了杀虫活性。分析质谱数据显示一些CTP与大片段相缔合以在这些样品中产生活性毒素复合物。样品3和4，即大片段和CTP单独测试；在所有情况下，这些样品在此单剂量时没有显示出与阴性对照显著不同的活性，这表明这两个片段都是需要的。

表13

胰蛋白酶处理的DIG-177样品1至4的抗西方玉米根虫的杀虫活性

*暴露于100μg/cm²后，一龄西部玉米根虫的平均幼虫致死率百分比和平均生长抑制百分比。阴性对照为350μg/cm²的Cry1Fa、10mM的CAPS(pH 10)和20mM柠檬酸钠(pH 3.5)缓冲液，而阳性对照为100μg/cm²的Cry34/35Ab1。

表14

TcdA、Cry34/35Ab1和DIG-1000的剂量响应

*值已经外推到测试的比率范围之外

实施例4

DIG-177的羧基末端的基因缺失

导致从DIG-177的羧基端的连续缺失的一系列五个编码区DIG-137(SEQ ID NO:53)、DIG-138(SEQ ID NO:55)、DIG-147(SEQ ID NO:57)、DIG-148(SEQ ID NO:59)和DIG-149(SEQ ID NO:61)如实施例2所述进行构建和表达。重组蛋白在假单胞菌(Pseudomonas)宿主细胞的可溶性级分中积累并如下纯化：将来自-80℃冷冻器的含重组蛋白的细胞糊转移至-20℃过夜，将其重悬于提取缓冲液(50mM Tris，5mM EDTA，pH 8.0)中。将其用匀化器充分混合，并使用具有30％占空比的平头电极(探针)的250(布兰森公司(Branson))超声波细胞粉碎仪进行5个循环的3分钟超声处理，随后每个循环后在冰上间断5分钟。细胞裂解后，将混合物在4℃以22,000x g离心25分钟。将上清液滤过0.45μm过滤器。纯化是使用离子交换和疏水相互作用树脂的两步色谱协定。首先，将上清液注入在50mM Tris，5mM EDTA，pH8.0中预平衡的Q柱(来自GE医疗集团，新泽西州皮斯卡塔韦的5mL HiTrap Q HP柱)中，并将蛋白质通过0-0.6M NaCl盐梯度洗脱。基于UV吸光度收集级分并通过SDS-PAGE进行检查。合并含有重组蛋白的级分并用缓冲液A加2M硫酸铵以1:1的比率稀释，最终的硫酸铵浓度达到1M。将样品施加到Phenyl HIC柱(来自GE医疗集团的5mL HiTrap Phenyl FF高取代柱)上，通过1-0M的(NH₄)₂SO₄梯度洗脱蛋白质。将含有靶蛋白的级分合并，并用10kDa MWCO离心过滤器浓缩，并用10mM COPS(pH10)渗析。通过Bradford和Densitometry测定蛋白质浓度。

如实施例3所述通过质谱确认蛋白质样品的组成。如实施例3所述测试材料的抗西方玉米根虫的杀虫活性。结果显示在测试浓度下，来自残基444-475(DIG-137)的区域缺失具有与DIG-177相似的活性。当用胰蛋白酶处理时DIG-137显示出降低的活性，而DIG-177如实施例3所示在未还原时保持活性。

此观察结果表明在没有胰蛋白酶处理的情况下CTP不是活性所特别需要的。更大的缺失(DIG-138、DIG-147、DIG-148和DIG-149)表现出比DIG-177对照更低的杀虫活性。对这些样品的胰蛋白酶处理使得杀虫活性降低超出未经胰蛋白酶处理的样品对WCR的杀虫活性。这些结果与Wei等人的那些结果一致，这显示Cry6Aa的直至残基382的羧基末端缺失在没有蛋白水解的情况下维持一些抗线虫动物的活性。这些结果证实了在残基390-475之间的区域中稳定化DIG-177多肽对保持有效的杀虫活性的重要性。

表14

抗WCR的羧基末端缺失的杀虫活性

*暴露于100μg/cm²的各种样品后，一龄西部玉米根虫的平均幼虫致死率百分比和平均生长抑制百分比，为两个数据组的平均值。阴性对照为350μg/cm²的Cry1Fa、10mM的CAPS(pH 10)和20mM柠檬酸钠(pH 3.5)缓冲液，而阳性对照为100μg/cm²的Cry34/35Ab1。

实施例5

对C88-C451二硫键的研究

为了研究cys88-cys451二硫键对杀虫活性的重要性，将突变体DIG-616(C163>A；(SEQ ID NO:39))和DIG-984(C88>A；451>A；(SEQ ID NO:49))如实施例2和3中所述进行鉴定。通过质谱法显示DIG-616和DIG-984蛋白具有预期的分子量和二硫键数量。

表15

在非还原和还原条件下对DIG-177突变体的完整分子量

分析。

在饮食生物测定中，DIG-616和DIG-984均在100ug/cm²时具有与DIG-177等同的抗WCR杀虫活性(表17)。当用胰蛋白酶处理时，DIG-616保持杀虫活性，而DIG-984当用胰蛋白酶处理时的杀虫活性低于DIG-616或DIG-177。质谱分析显示胰蛋白酶处理的DIG-616保留CTP而胰蛋白酶处理的DIG-984丧失了CTP，这证实了蛋白水解后抗西方玉米根虫的活性需要该肽(表16)，以及在缺乏二硫键的情况下LF和CTP的有限亲和性。

表16

在非还原和还原条件下在用胰蛋白酶消化后对DIG-177突变体的完整分子量

分析

表17

具有和不具有抗WCR的胰蛋白酶处理的DIG-177及其突变体的

杀虫活性

*暴露于100μg/cm²的各种样品后，一龄西部玉米根虫的平均幼虫致死率百分比和平均生长抑制百分比。阴性对照为350μg/cm²的Cry1Fa、10mM的CAPS(pH10)和20mM柠檬酸钠(pH 3.5)缓冲液，而阳性对照为100μg/cm²的DIG-177。

实施例6

DIG-177的蛋白酶K消化

使用蛋白酶K确定DIG-177蛋白的蛋白水解易感性。(蛋白酶K已显示具有比胰蛋白酶更宽的特异性，在脂肪族和芳香族残基的羧基侧切割，而胰蛋白酶在赖氨酸和精氨酸残基处切割)。如实施例2中所述制备DIG-177样品蛋白质。使用20℃下的轻微摇动将该样品蛋白质以浓度1mg/mL在50mM Tris(pH 7.5)、2mM CaCl₂中以与蛋白酶K(西格玛奥德里奇公司，密苏里州圣路易)40:1的质量比消化。在0、5、10、30、50和90分钟取得时间点样品，并通过加入PMSF至最终浓度5mM来终止消化。蛋白酶K加入样品后立即取0时间点。如实施例2和3所述，通过SDS-PAGE、质谱和昆虫生物测定分析样品。

在这些条件下将全长DIG-177消化至50分钟，从而产生了约43kDa的大片段。由于蛋白酶K的广泛特异性，所以肽端点在残基5-73的氨基末端和残基386-456的羧基末端处具有不均匀范围。取决于肽的端点，在某些情况下，CTP片段通过C88-C451二硫键与大片段连接。发现具有易感性的蛋白酶K区与实施例3中呈现的胰蛋白酶结果一致。

实施例7

针对杀虫活性和蛋白酶K稳定性堆DIG-177内部缺失

进行表征

对DIG-177的蛋白水解加工区域，SEQ ID NO:的残基约390-443处进行三个系列的序列缺失。2.第一个缺失组是5个氨基酸，从而产生了蛋白质(DIG-921至927(SEQ ID NO:64、SEQ ID NO:66、SEQ ID NO:68、SEQ ID NO:70、SEQ ID NO:72、SEQ ID NO:74、SEQ IDNO:76)和DIG-931(SEQ ID NO:78)，第二个缺失组是10个氨基酸(DIG-969至973(SEQ IDNO:80、SEQ ID NO:82、SEQ ID NO:84、SEQ ID NO:86、SEQ ID NO:88))，第三个缺失组(DIG-985至997(SEQ ID NO:90、SEQ ID NO:92、SEQ ID NO:94、SEQ ID NO:96、SEQ ID NO:98、SEQID NO:100、SEQ ID NO:102、SEQ ID NO:104、SEQ ID NO:106、SEQ ID NO:108、SEQ ID NO:110、SEQ ID NO:112、SEQ ID NO:114))是15、20、25和43个残基。如实施例2和3中所述，每个系列在假单胞菌属中表达、纯化，并测试其抗西方玉米根虫的杀虫活性。如上所述通过用蛋白酶K消化进行消化来测定蛋白质的稳定性。在所有情况下，突变蛋白都具有杀虫活性。这些蛋白质与DIG-177几乎一样对蛋白酶K敏感；一个明显的例外是DIG-995(SEQ ID NO:110)，其为表现出更少消化的一个43个氨基酸的缺失。

表18

DIG-177缺失的说明

实施例8

经胰蛋白酶处理的DIG-177的晶体结构测定

包涵体(IB)的制备.将表达全长DIG-177(SEQ ID NO:2)的荧光假单胞菌来源的细胞糊从-80℃贮藏转移至4℃并以20％w/v重悬于冷裂解缓冲液(50mM Tris、200mM NaCl、10％甘油、0.5％Triton X-100、20mM EDTA、1mM TCEP、pH 7.5)中，并用手持匀化器充分混合。然后使悬浮液以16,000psi穿过微流化器(M-110EH)两次，然后离心(SLC-6000转子/14,000g/40分钟/4℃)。弃去上清液。将包涵体沉淀重悬于含有0.4g/L溶菌酶(L-6876；西格玛奥德里奇公司)的10％w/v室温裂解缓冲液中并通过匀化完全重悬。将悬浮液在30℃温育30分钟，每10分钟进行一次简单的匀化。然后将包涵体离心(SLC-6000转子/14,000g/40分钟/4℃)并弃去上清液。使用匀化器将沉淀物在10％w/v冷裂解缓冲液中重悬浮最后一次并离心(SLC-6000转子/14,000g/40分钟/4℃)，然后弃去上清液。使用匀化器在不含Triton X-100的冷裂解缓冲液(50mM Tris、200mM NaCl、10％甘油，20mM EDTA，1mM TECP，pH 7.5)中以10％w/v悬浮包涵体沉淀并离心(SLC-6000转子/14,000g/40分钟/4℃)，然后弃去上清液；重复此步骤。将包涵体在10mM EDTA(pH 8.0)中重悬至30％(w/v)，或等分成1.5mL等分试样并在-80℃冷冻直至需要时。

DIG-177胰蛋白酶核心的制备.八支2ml试管的全长DIG-177的IB悬浮液解冻并在80ml最终体积(1:5稀释)的10mM CAPS(pH 11.0)中提取。pH值测得为9.1，并随后用NaOH调节至11.0。向提取物中加入50mg新鲜制备的胰蛋白酶(在(1mM HCl、5mM CaCl₂)中的25mg/ml胰蛋白酶溶液(西格玛公司(Sigma)，用T1426-1G TLCK处理过的))。将pH保持在11.0并在4℃搅拌过夜。

将该溶液以10mL/分钟的流速施加到在缓冲液A(25mM CAPS，pH 11.0)中预平衡的Source 15Q 16/10柱中，并用缓冲液A+1M NaCl梯度洗脱75分钟。截短的DIG-177作为单个大峰洗脱，将其使用四个15ml Amicon 10,000MWCO旋转浓缩器在4℃和5000x g下的JA-12转子中从100mL浓缩至10mL。将浓缩的离子交换样品施加至在25mM CAPS(pH 11.0)和50mMNaCl中预平衡的Superdex 7526/90凝胶过滤柱。将样品使用2.5mL/分钟的流速洗脱。合并包含在大DIG-177核心峰中的级分(80mL，2.05mg/mL)。

全长DIG-177的制备.将约500mg的IB在4℃解冻，并在4°以23,000x g离心25分钟。去除上清液并将IB沉淀在30mL的100mM CAPS(pH 11.0)中溶解，并在室温下温和摇动悬浮液小时以溶解DIG-177蛋白。溶解后，将混合物在4°以23,000x g离心25分钟。将上清液用25mM CAPS(pH 10.0)渗析过夜。

然后将经缓冲液交换的DIG-177样品滤过0.22微米注射器式过滤器并以5mL/min流速施加至Source 15Q 16/6离子交换柱。将柱用缓冲液A(25mM CAPS，pH 10)预平衡.用20％、30％、40％和50％的缓冲液B(缓冲液A+1M NaCl)的不连续梯度洗脱蛋白质。每次洗脱液为约50mL。基于紫外吸光度从各个洗脱液收集级分并通过SDS-PAGE进行分析。

全长DIG-177在20％B级分中，将该级分合并和使用具有10kDa分子量截留膜(密理博公司)的离心过滤装置浓缩至约30mL。

将浓缩的样品通过尺寸排阻色谱进一步纯化。对于每次跑样，将4.0mL样品以2.5mL/min的流速施加于在25mM CAPS(pH 10)、50mM NaCl缓冲液中预平衡的Superdex 7526/90凝胶过滤柱上。观察到了两个峰值；第一个峰在空隙体积处洗脱并含有DIG-177二聚体。来自峰2的级分主要含有单体。将来自峰2的级分分别合并并提交进行结晶实验。

蛋白质浓度测定.根据制造商的说明进行BCA测定(皮尔斯生命技术公司(PierceLife Technologies)，纽约州格兰德岛)，除了将1000μL工作试剂加入到50μl样品中之外。根据制造商的说明进行Bradford蛋白质测定(伯乐公司，美国加州赫拉克勒斯)，除了将1000μL工作试剂加入到20μL样品中之外。

DIG-177胰蛋白酶核心的结晶、数据收集和结构测定.将DIG-177胰蛋白酶核心浓缩至100mg/mL。初始晶体使用Rigaku Reagents公司的Wizard Classic I(华盛顿州班布里奇岛)获得。在筛选多个条件后，从20％(w/v)PEG 1000、0.1M磷酸钠/柠檬酸，pH 4.2；0.2M硫酸锂获得合适的晶体。在LS-CAT(先进质子来源(Advanced Proton Sources)，阿贡国家实验室(Argonne National Laboratory))的Mar CCD-300检测器上以100K从单晶收集数据。细胞常数为a＝112.07，b＝112.07，c＝76.6，α＝90.0，β＝90.0，γ＝120.0。家庭来源的最初数据收集建议空间群为P6₅或P6₁；因此，数据最初是在P6₅中处理的。然而，后续分析导致认识到空间群是P6₃。

DIG-177毒素的截短形式的结构通过使用PHASER(1)(CCP4包(Winn,M.D.等人，2011))的分子置换法随后进行手工重建和模型优化来解析。将由残基19-334组成的来自蜡状芽孢杆菌(Bacillus cereus)的溶血素B的晶体结构的聚丙氨酸链(蛋白质数据库条目2NRJ)用作搜索模型。通过进行由使用COOT(Emsley等人，2010)的手动模型构建组成的若干个精化循环，然后用REFMAC(Murshudov等人，1997)进行约束精化，来获得最终模型。

表19

经胰蛋白酶处理的

DIG-177的结晶数据收集和细化统计(PDB:)。

^aR_合并＝100Σ(h)Σ(i)|I(i)-<I>|/Σ(h)Σ(i)I(i)，其中I(i)是反射h的第i次强度测量值，并且<I>是来自多次观测的平均强度。

^bR_结晶＝Σ||F_obs|-|F_calc||/Σ|F_obs|。其中F_obs和F_calc分别是来自数据和模型的结构因子幅值。R_游离是具有结构因子的10％的R_结晶。

全长DIG-177的结晶、数据收集和结构测定在10mM HEPES缓冲液，pH 7.5和25mMNaCl中使用具有10kDa分子量截留(密理博公司)的Amicon离心过滤器将全长DIG-177蛋白浓缩至15mg/ml。使用市售的Classics、Classics Lite、Classics II、PEG's、PEG's II、PhClear和PACT筛选(汉普顿研究中心(Hampton Research)，加利福尼亚州亚里索维耶荷)，通过使用蚊式机器人系统(TTP LabTech公司，英国赫特福德郡)在96圆底孔结晶板(葛莱娜公司(Greiner Bio-One，GmbH)，德国)中进行沉滴方法，来执行初始结晶筛选。衍射质量DIG-177蛋白质晶体在291K下从含有3μL蛋白质样品和1.5μL贮存液(0.1M柠檬酸，pH 4.6，4％PEG 6,000)的沉滴生长。通过将晶体溶解在SDS缓冲液中获得的蛋白质样品的SDS-PAGE分析未揭示任何降解产物，并确认在用于数据收集的3s中仅存在全长DIG-177蛋白质。

为了数据收集，在贮存液中用20％(v/v)甘油收获晶体。在LS-CAT(先进质子来源(Advanced Proton Sources)，阿贡国家实验室(Argonne National Laboratory))的MarCCD-300检测器上以100K从单晶收集衍射数据。使用HKL-2000(Z.Otwinowski和W.Minor，1997)对数据进行索引和处理。晶体属于正交空间群P2₁2₁2，并且每个不对称单元含有一个全长DIG-177分子。

全长DIG-177的结构通过使用PHASER(McCoy,A.J.J.Appl.Cryst.(2007))，以DIG-177的截短形式的结构作为搜索模型进行分子置换来解析。通过进行由使用COOT(Winn,M.D.等人，2011)的手动模型构建组成的若干个循环，然后用REFMAC(Murshudov等人，1997)进行结构精化，来获得全长DIG-177毒素的最终模型。

表20

全长DIG-177的数据收集和精化统计

(PDB：Ronda以检查提交)

^aR_合并＝100Σ(h)Σ(i)|I(i)-<I>|/Σ(h)Σ(i)I(i)，其中I(i)是反射h的第i次强度测量值，并且<I>是来自多次观测的平均强度。

^bR_结晶＝Σ||F_obs|-|F_calc||/Σ|F_obs|。其中F_obs和F_calc分别是来自数据和模型的结构因子幅值。R_游离是具有结构因子的10％的R_结晶。

胰蛋白酶处理的DIG-177和全长DIG-177的分子结构.图2中示出的经胰蛋白酶处理的Cry6Aa(DIG-177)的分子结构的带状图由α螺旋束核心和折叠到该束上的α螺旋发夹结构组成。在结构上DIG-177被识别为α螺旋溶血素并且与大肠杆菌溶血素E(1QOY；Wallace2000)和来自蜡状芽孢杆菌的溶血素BL的B组分(2NRJ；Madegowda等人，2008)共享结构相似性。

全长DIG-177的分子结构与胰蛋白酶处理的结构几乎可重叠；125-128与387-451之间的残基未被解析；387-452之间的残基被模型化。这些结构示出于图2中。

实施例9

蛋白水解稳定的DIG-177变体的构建

Cry6Aa(DIG-177)已显示是来自苏云金芽孢杆菌的晶体蛋白，其在饮食生物测定中对西方玉米根虫(Diabrotica virgifera virgifera)具有杀虫活性。大约在残基390-451之间的区域中的蛋白水解导致杀昆虫活性的降低。为了限制DIG-177的蛋白水解敏感性，使用3D分子结构来设计此区域的替换接头。用接头替代的片段在残基381和457之间使用若干个端点。一些接头使用购自化学计算集团(Chemical Computing Group)(加拿大魁北克省蒙特利尔)的MOE(分子操作环境)软件的循环建模器功能来建模。

如实施例4和6中所述，新设计在假单胞菌中表达、纯化，并测试其杀虫活性和相对蛋白酶K抗性。这项工作表明，相较于亲本蛋白DIG-177，若干种变体具有杀虫活性和增加的蛋白酶K抗性。

表21

测试DIG-177变体以提供增加的蛋白酶抗性

接头序列示出在斜线标记之间。对西方玉米根虫的杀虫活性显示为致死率％，并且使用实施例6中描述的蛋白酶K测定法测定相对蛋白酶K抗性(0＝与DIG-177相似的蛋白酶K敏感性；+＝相对于DIG-177增加的蛋白酶K抗性)。--是DIG-177蛋白内的缺失。

实施例10

DIG-1000的鉴定

制备DIG-1000(SEQ ID NO116)，一种已经用7残基接头：VATITSG将残基苏氨酸387替换成脯氨酸452的Cry6Aa的新型变体。使用DIG-177胰蛋白酶核心的3D晶体结构和购自化学计算集团(加拿大魁北克省蒙特利尔)的MOE(分子操作环境)软件的循环建模器功能来选择接头区域。该接头被设计为通过用较短的较不易感的区段代替较大的易感环来限制蛋白水解(参见图3)。

如实施例3中所述，DIG-1000在假单胞菌中表达、纯化和生物测定。已显示DIG-1000对西方玉米根虫具有杀虫活性。

表22

DIG-177和DIG-1000的抗西方玉米根虫的杀虫活性

蛋白酶K抗性.如实施例3所述表达和纯化DIG-177和DIG-1000。如实施例6中那样测试蛋白质对蛋白酶K抗性的易感性(图4)。DIG-177(54.2kDa)在T＝0时间点(加入蛋白酶，样品立即取出)表现出显著的部分降解。继续降解直到在90分钟处观察到蛋白水解稳定的核心。相比之下，DIG-1000(47.3kDa)在T＝0时不显示降解，在T＝5分钟时观察到一些部分降解，在10分钟时约50％的全长蛋白质被转化。转化为蛋白水解稳定的核心在50分钟处完成。这些数据证实DIG-1000在体外对蛋白酶K具有更强的抗性。

实施例11

DIG-1000的瞬时玉米表达

使用授粉后10-12天收获的未成熟玉蜀黍(B104)胚胎的粒子轰击测试DIG-177和DIG-1000的瞬时表达(WO 2014/028295A；US 2012/0060238Al)。如实施例1中所述，重构DIG-177(SEQ ID NO:1)和DIG-1000(SEQ ID NO:115)的编码区以反映在分别产生SEQ IDNO:3和SEQ ID NO:137的玉蜀黍细胞中的瞬时测试的玉蜀黍密码子偏倚。还在DIG-177和DIG-1000上测试用以将表达的蛋白质引导到叶绿体隔室中的转运肽(TraP)。修饰的编码区分别示出于SEQ ID NO:32和SEQ ID NO:139中，以分别产生多肽SEQ ID NO:33和SEQ IDNO:140。

表达来自如表23所示含有玉蜀黍泛素启动子、所关注的编码区和玉蜀黍过氧化物酶5 3’非翻译区的基于pUC的质粒。使用黄色荧光蛋白和PAT(草丁膦乙酰转移酶)表达质粒作为表达对照。

表23

用于玉蜀黍胚胎瞬时表达的质粒

通过将50mg的0.6μm或1μm金微载体(伯乐公司，美国加州赫拉克勒斯)称入无菌2.0ml微量离心管中制备金颗粒贮存液。将颗粒用乙醇洗涤三次，随后用无菌水洗涤三次，在每次洗涤后，通过在微量离心机中以1500g旋转2分钟来收集物质。将颗粒悬浮于500μl无菌的50％甘油中并贮藏于-20℃。

使用表面消毒的B104未成熟穗来分离胚胎。从3-4个穗中将未成熟胚胎(授粉后10-12天；1.8-2.4mm)分离到含有1.75ml液体或2.2-4.3g/L MS盐(Murshige和Skoog，1962)和1ml/l改性的MS维生素溶液(1000x)或4.3LS培养基(Linsmaier和Skoog，1965)和1ml/lChu N6维生素溶液(1000x)、68.4g/l蔗糖、36g/l葡萄糖、100-700mg/l L，以及有或没有1.5mg/l的2,4-D(2,4-二氯苯氧乙酸)的2ml微量离心管中。

胚胎分离后，取出液体培养基并丢弃。然后将胚胎培养到含有用于渗透处理的半固体培养基的培养皿上。此培养基由4.3g/l MS盐、1ml/l改性的MS维生素溶液(1000x)、500mg/l MES、100mg/l肌醇，100mg/l酪蛋白酶促水解产物或120g/l蔗糖或45g/l的各种山梨糖醇和甘露糖醇、以及含或不含3.3mg/l麦草畏、15mg/l硝酸银和2.5g/l凝胶糖(结冷胶)组成。将胚胎在目标区域内以5×8的方格排列以进行粒子轰击。在进行粒子轰击之前，将所有平板在27℃下于24小时处于50μM低光下的腔室温育24小时。

在涂布金颗粒之前，将来自所关注构建体的每种DNA与包含标准基因PAT的对照构建体以1:1的比率混合。另外，将来自含有YFP基因的构建体的DNA用于转化作为视觉对照。将含有来自上面的50μl金颗粒贮存液的各个管悬浮到无菌的2.0ml管中。将以下物质添加到每个金颗粒管中：pDAB112364的测试构建体DNA(5μg)/对照构建体DNA(5μg)、50μl的2.5MCaCl₂和20μl的0.1M亚精胺。然后将管在室温下高速涡旋10-15分钟。接着用200μl的100％乙醇洗涤三次，最后将包被的金颗粒重悬于30μl的100％乙醇中，并将所有管置于冰上。

标记每个测试构建体的两个大载体，并将5μl的DNA/金混合物均匀地散布在大载体的中央，并使其干燥10分钟。将范围在650-1100psi之间的爆破膜(伯乐公司，美国加州赫拉克勒斯)用70％丙醇灭菌并在轰击前部分干燥。轰击含有每个构建体的40个胚胎的两个平板。将经轰击的胚胎保持在相同的培养基上并在27℃下在50μM低光照条件下温育24小时过夜。

轰击24小时后，观察转化的胚胎与对照构建体pDAB100286的YFP表达。此对照用于监测DNA/金涂布过程以及粒子轰击过程。在确认对照胚胎中的YFP表达后，对来自每个测试构建体的两块平板取样以进行蛋白质分析。每个样品含有20个胚胎，以允许多个技术平行测定用于蛋白质分析以及转化过程。每个构建体总共四个样品被提交用于蛋白质分析。

在轰击和温育之后，将样品贮藏在-80°的96孔集束式管架中，直到分析当天。将两个Daisy^TM钢BB和300μl提取缓冲液(含有0.05％吐温20和5μl/ml西格玛蛋白酶抑制剂的PBS溶液，产品目录号9599)加入到每个管中。在最大设定下将样品在Kelco珠磨机中研磨3分钟。将样品以3,000x g离心5分钟；将100μl上清液转移到空样品管中。

常规电泳和印迹(Gallagher,S.等人，2008)方法与Invitrogen^TM装置和基本试剂一起使用。兔抗Cry6a抗体是用于检测Cry6a的一抗。所有蛋白质用Cy3荧光检测系统检测，并使用GE Typhoon^TM成像系统(GE医疗集团，新泽西州皮斯卡塔韦)进行扫描。

表达实验的结果示出于图5中。蛋白印迹分析显示在阴性对照中没有表达Cry6Aa蛋白，所述阴性对照包括未轰击的玉蜀黍胚胎，以及用YFP和PAT质粒轰击的那些。两条最右边的泳道是DIG-177的蛋白质标准品、全长Cry6Aa(54.1kDa)和DIG-1000(47.3kDa)。DIG-177+TraP泳道含有高于标准品的模糊条带，可能代表前体蛋白。还有三条其他条带，其中一个条带在大小上接近DIG-177标准品，而其他条带显著较小，可能指示部分降解；DIG-177泳道表现出类似的模式，但没有推定的前体蛋白质。DIG-1000和DIG-1000+TraP看起来与看似用DIG-1000标准品迁移的单一条带相同(考虑到凝胶末端泳道的一些变形)。这些数据与表达可用于西方玉米根虫控制的植物细胞稳定的杀虫蛋白的DIG-1000编码序列一致。

实施例12

DIG-1000杀虫毒素的稳定双子叶植物表达

拟南芥转化.用花序浸渍法(Weigel和Glazebrook，2002)转化拟南芥Col-01。选择的农杆菌菌落用于接种1mL至15mL含有适当抗生素的YEP肉汤培养物以进行选择。将培养物在28℃下用220rpm恒速搅拌温育过夜。每个培养物用于接种两个含有适当抗生素的500mLYEP肉汤培养物以进行选择，并且将新培养物在28℃下用恒温搅拌温育过夜。将细胞在室温下以约8700x g沉淀10分钟，并弃去所得上清液。将细胞沉淀温和地重悬于500mL含有以下物质的浸润培养基中：1/2x Murashige和Skoog盐(西格玛奥德里奇公司)/Gamborg的B5维生素(Gold BioTechnology公司，密苏里州圣路易)、10％(w/v)蔗糖、0.044μM苄基氨基嘌呤(10μL/升的1mg/mLDMSO贮存液)和300μL/升的Silwet L-77。将大约1个月大的植株浸入培养基中15秒，小心确保浸没最新花序。然后将植株以它们的侧面放置并加盖(透明或不透明)24小时，用水清洗并直立放置。植株在22℃，使用16小时光照/8小时黑暗光周期来生长。浸渍大约4周后，收获种子。

拟南芥属生长和选择.使新鲜收获的T1种子在室温下在干燥剂存在下干燥至少7天。将种子悬浮于0.1％琼脂/水(西格玛奥德里奇公司)溶液中，然后在4℃下分层2天。为准备种植，将10.5英寸×21英寸发芽盘(TO Plastics公司，华盛顿州克利尔沃特)中的Sunshine Mix LP5(Sun Gro Horticulture公司，华盛顿州贝尔维尤)用精细蛭石覆盖，用Hoagland溶液(Hoagland和Arnon，1950)自吸式底部灌溉(sub-irrigated)直到潮湿，然后排水24小时。将分层的种子播种到蛭石上并用保湿盖(humidity dome)(KORD Products公司，加拿大安大略省布兰普顿)覆盖7天。使种子萌发并且在Conviron^TM生长腔室(CMP4030和CMP3244型；Controlled Environments Limited公司，加拿大马尼托巴省温尼伯)中在长光照条件(16小时光照/8小时黑暗)下以120-150μmol/m²sec的光强度在恒温(22℃)和恒湿(40-50％)下培养植物。最初用霍格兰溶液浇灌植物，然后用去离子水浇灌以保持土壤潮而不湿(moist but not wet)。

播种后5-6天去除保湿盖，并用化学选择剂喷洒植物以杀死从非转化种子发芽的植物。例如，如果由二元植物转化载体提供的植物可表达选择标记基因是pat或bar基因(Wehrmann等人，1996)，则可以通过喷雾1000X Finale溶液(5.78％草铵膦，FarnamCompanies公司，亚利桑那州菲尼克斯)来选择转化的植物。两次后续喷雾以5-7天的间隔进行。在最后一次喷雾后7-10天确定存活者(植物有活性地生长)并移植到用Sunshine MixLP5制备的盆中。将移植植物用保湿盖覆盖3-4天，并置于在上述生长条件下的Conviron^TM生长腔室中。

双子叶植物转化领域的技术人员将会理解，当使用其他植物可表达的选择标记基因(例如除草剂耐受性基因)时，其他选择转化植物的方法是可用的。

转基因拟南芥的抗虫性生物测定.表达DIG-1000杀虫毒素蛋白的转基因拟南芥品系被证实在人工饮食覆盖测定中对敏感昆虫物种有活性。通过适当的方法定量从转基因和非转基因拟南芥品系中提取的蛋白质，并调整样品体积以使蛋白质浓度归一化。如上所述在人工饮食上进行生物测定。测定中包括非转基因拟南芥和/或缓冲液和水作为背景检查处理。

实施例13

DIG-1000杀虫毒素的稳定玉蜀黍表达

农杆菌介导的玉蜀黍转化。将来自High II F1杂交的种子(Armstrong等人，1991)种植到含有95％Metro-Mix 360无土生长培养基(Sun Gro Horticulture公司，华盛顿州贝尔维尤)和5％的粘土/壤质土的混合物的5加仑盆中。植物在使用高压钠灯和金属卤化物灯的组合、具有16:8小时光照:黑暗光周期的温室中生长。为了获得未成熟的F₂胚胎以进行转化，进行受控的近缘授粉。在授粉后8-10天，当胚胎大小为约1.0至2.0mm时，分离未成熟胚胎。

感染和共培养.通过用液体肥皂擦洗、在70％乙醇中浸泡2分钟，然后将其浸入20％市售漂白剂(0.1％次氯酸钠)中30分钟，之后用无菌水冲洗，来对玉蜀黍穗进行表面消毒。通过将在含有100mg/L壮观霉素、10mg/L四环素和250mg/L链霉素的YEP固体培养基上于28℃下生长2-3天的1-2个环细菌转移到含有100μM乙酰丁香酮的5mL液体感染培养基(LS基础培养基(Linsmaier和Skoog，1965)、N6维生素(Chu等人，1975)、1.5mg/L的2,4-二氯苯氧乙酸(2,4-D)、68.5gm/L蔗糖、36.0gm/L葡萄糖、6mM L脯氨酸，pH 5.2)来制备含有超二元载体的悬浮农杆菌细胞。将溶液涡旋搅拌直至获得均匀的悬浮液，并使用具有紫色过滤器的Klett-Summerson色度计将浓度调节至200Klett单位的最终密度或在600nm测量的等效光密度(OD₆₀₀)。将未成熟的胚胎直接分离到含有2mL感染培养基的微量离心管中。去除培养基并用1mL密度为200Klett单位或等同的OD₆₀₀的农杆菌溶液替换，并将农杆菌和胚胎溶液在室温下温育5分钟，然后转移到共培养培养基(LS基础培养基、N6维生素、1.5mg/L 2,4-D、30.0gm/L蔗糖、6mM L脯氨酸、0.85mg/L AgNO₃、100μM乙酰丁香酮，3.0gm/L结冷胶(PhytoTechnology Laboratories.公司，堪萨斯州雷那克萨市)，pH 5.8)在25℃于黑暗条件下5天。

共培养后，将胚胎转移至选择培养基中，然后在约8周的时间内获得转化的分离物。为了选择用含有植物可表达的pat或bar选择标记基因的超二元质粒转化的玉蜀黍组织，将基于LS的培养基(LS基础培养基、N6维生素、1.5mg/L的2,4-D、0.5gm/L的MES(2-(N-吗啉代)乙磺酸一水合物；PhytoTechnologies Labr.)、30.0gm/L蔗糖、6mM L脯氨酸、1.0mg/LAgNO₃、250mg/L头孢噻肟、2.5gm/L结冷胶，pH 5.7)与双丙氨膦(Gold BioTechnology公司)一起使用。将胚胎转移至含有3mg/L双丙氨膦的选择培养基中，直至获得胚胎发生分离物。回收的分离物通过以2周的间隔转移到新鲜选择培养基中壮大(bulked up)以进行再生和进一步分析。

玉蜀黍转化领域的技术人员将理解，当使用其他植物可表达的选择标记基因(例如除草剂耐受性基因)时，其他选择转化植物的方法是可用的。

再生和种子生产.为了再生，将培养物转移至“28”诱导培养基(MS盐和维生素，30gm/L蔗糖、5mg/L苄氨基嘌呤、0.25mg/L 2,4-D、3mg/L双丙氨膦、250mg/L头孢噻肟、2.5gm/L结冷胶，pH 5.7)在低光照条件(14μEm^-2s^-1)下1周，然后在高光照条件(约89μEm^-2s^-1)下1周。随后将组织转移到“36”再生培养基(除了缺乏植物生长调节剂外与诱导培养基相同)。当小植株长到3-5厘米长时，将它们转移到含有SHGA培养基(Schenk和Hildebrandt(1972)盐和维生素)；PhytoTechnologies Labr.)、1.0gm/L肌醇、10gm/L蔗糖和2.0gm/L结冷胶，pH 5.8)的玻璃培养管中，以允许芽和根的进一步生长和发育。如本文前面所述将植物移植到相同的土壤混合物中，并在温室中生长至开花。进行用于种子生产的受控授粉。

实施例14

转基因玉蜀黍的生物测定

通过常规生物测定方法证明表达在植物细胞中产生的本发明的杀虫毒素的稳定转化植物的生物活性(参见例如Huang等人，2006)。技术人员能够例如通过在受控的饲喂环境下给靶标昆虫喂食来源于产生工程化的Cry6Aa杀虫毒素的植物的各种植物组织或组织片来证实效力。或者，蛋白质提取物可以从来源于产生工程化的Cry6Aa杀虫毒素的植物的各种植物组织制备，并且将提取的蛋白质掺入如本文先前描述的人工饮食生物测定中。应当理解，将这样的饲喂测定的结果与类似进行的采用来自不产生工程化的Cry6Aa杀虫毒素的宿主植物的适当对照组织、或其他对照样品的生物测定进行比较。

参考文献

Alistair J.Wallace,Timothy J.Stillman,Angela Atkins,StuartJ.Jamieson,Per A.Bullough,Jeffrey Green,and Peter J.Artymiuk(2000).“E.coliHemolysin E(HlyE,ClyA,SheA):X-Ray Crystal Structure of the Toxin andObservation of Membrane Pores by Electron Microscopy”.Cell,Vol.100:265–276,January Altschul,S.F.,Gish,W.,Miller,W.,Myers,E.W.,Lipman,D.J.(1990)Basiclocal alignment search tool.J.Mol.Biol.215:403-410.

Altschul,S.F.,Madden,T.L.,A.A.,Zhang,J.,Zhang,Z.,Miller,W.,Lipman,D.J.(1997)Gapped BLAST and PSI-BLAST:a new generation of proteindatabase search programs.Nucl.Acids Res.25:3389-3402.

An,G.,Watson,B.D.,Stachel,S.,Gordon,M.P.,Nester,E.W.(1985)New cloningvehicles for transformation of higher plants.EMBO J.4:277–284.

Armstrong,C.L.,Green,C.E.,Phillips,R.L.(1991)Development andavailability of germplasm with high TypeII culture formation response.MaizeGenet.Coop.Newslett.65:92-93.

Aronson,A.I.,Han,E.-S.,McGaughey,W.,Johnson,D.(1991)The solubility ofinclusion proteins from Bacillus thuringiensis is dependent upon protoxincomposition and is a factor in toxicity to insects.Appl.Environ.Microbiol.57:981-986.

Aronson,A.I.,Geng,C.,Wu.L.(1999)Aggregation of Bacillus thuringiensisCry1A toxins upon binding to target insect larval midgutvesicles.Appl.Environ.Microbiol.65:2503-2507.

Arvidson,H.,Dunn,P.E.,Strand,S.,Aronson,A.I.(1989)Specificity ofBacillus thuringiensis for lepidopteran larvae:factors involved in vivo andin the structure of a purified toxin.Molec.Microbiol.3:1533-1543.

Ausubel et al.,eds.(1995)Current Protocols in Molecular Biology,Chapter 2(Greene Publishing and Wiley-Interscience,New York).

Bailey,J.M.,Shenov,N.R.,Ronk,M.,and Shively,J.E.,(1992)Automatedcarboxy-terminal sequence analysis of peptides.Protein Sci.1:68–80.

Baum JA,Bogaert T,Clinton W,Heck GR,Feldmann P,Ilagan O,Johnson S,Plaetinck G,Munyikwa T,Pleau M,Vaughn T,Roberts J.,(2007)Control ofcoleopteran insect pests through RNA interference.Nat Biotechnol.Nov；25(11):1322-6.Epub 2007 Nov 4.

Beltz,G.A.,Jacobs,K.A.,Eickbush,T.H.,Cherbas,P.T.,Kafatos,F.C.(1983)Isolation of multigene families and determination of homologies by filterhybridization methods.InWu,R.,Grossman,L.,Moldave,K.(eds.)Methods ofEnzymology,Vol.100 Academic Press,New York pp.266-285.

Benchabane,M.,Goulet,C.,Rivard,D.,Faye,L.,Gomord,V.,Michaud,D.,(2008).“Preventing Unitended Proteolysis in Plant Protein Biofactories”.PlantBiotechnology Journal 6:633-648.

Bown,D.P.,Wilkinson,H.S.,Jongsma,M.A.,Gatehouse,J.A.(2004)Characterization of cysteine proteinases responsible for digestiveproteolysis in guts of larval western corn rootworm(Diabrotica virgifera)byexpression in the yeast Pichia pastoris.Insect Biochem.Molec.Biol.34,:305-320.

Bravo,A.,Gill,S.S.,Soberon,M.(2007)Mode of action of Bacillusthuringiensis Cry and Cyt toxins and their potential for insectcontrol.Toxicon49:423-435.

Caruthers,M.H.,Kierzek,R.,Tang,J.Y.(1987)Synthesis ofoligonucleotides using the phosphoramidite method.Bioactive Molecules(Biophosphates Their Analogues)3:3-21.

Cervelli,M.,Tavladoraki,P.,Agostino,S.,Angelini,R.,Rodolfo,F.,andMariottini,P.(2000).“Isolation and characterization of three polyamineoxidase genes from Zea mays”.Plant Physiol.Biochem.38:667-677.

Christeller,J.T.,Laing,W.A.,Markwick,N.P.,Burgess,E.P.J.(1992)Midgutprotease activities in 12 phytophagous lepidopteran larvae:dietary andprotease inhibitor interactions.Insect Biochem.Molec.Biol.22:735-746.

Chu,C.C.,Wand,C.C.,Sun,C.S.,Hsu,C.,Yin,K.C.,Chu,C.Y.,Bi,F.Y.(1975)Establishment of an efficient medium for anther culture of rice throughcomparative experiments on the nitrogen sources.Scientia Sinica 18:659-668.

Crameri,A.,Cwirla,S.,Stemmer,W.P.C.(1996a)Construction and evolutionof antibody-phage libraries by DNA shuffling.Nat.Med.2:100-103.

Crameri,A.,Whitehom,E.A.,Tate,E.,Stemmer,W.P.C.(1996b)Improved greenfluorescent protein by molecular evolution using DNAshuffling.Nat.Biotech.14:315-319.

Crameri,A.,Dawes,G.,Rodriguez,E.,Silver,S.,Stemmer,W.P.C.(1997)Molecular evolution of an arsenate detoxification pathway by DNAshuffling.Nat.Biotech.15:436-438.

Crickmore N.,Zeigler,D.R.,Feitelson J.,Schnepf,E.,Van Rie J.,LereclusD.,Baum J.,and Dean D.H.(1998)Revision of the Nomenclature for the Bacillusthuringiensis Pesticidal Crystal Proteins Microbiol.Mol.Biol.Reviews62:807-813.

de Maagd,R.A.,Kwa,M.S.,van der Klei,H.,Yamamoto,T.,Schipper,B.,Vlak,J.M.,Stiekema,W.J.,Bosch,D.(1996)Domain III substitution in Bacillusthuringiensis delta-endotoxin CryIA(b)results in superior toxicity forSpodoptera exigua and altered membrane proteinrecognition.Appl.Environ.Microbiol.62:1537-1543.

de Maagd,R.A.,Bravo,A.,Berry,C.,Crickmore,N.,Schnepf,E.(2003)Structure,diversity,and evolution of protein toxins from spore-formingentomopathogenic bacteria.Annu.Rev.Genet.37:409-433.

Diaz-Mendoza,M.,Farinos,G.P.,Castanera,P.,Hernandez-Crespo,P.,Ortego,F.(2007)Proteolytic processing of native Cry1Ab toxin by midgut extracts andpurified trypsins from the Mediterranean corn borer Sesamianonagrioide.J.Insect Physiol.53:428-435.

Ellis,R.T.,Stockhoff,B.A.,Stamp,L.,Schnepf,H.E.,Schwab,G.E.,Knuth,M.,Russell,J.,Cardineau,G.A.,Narva,K.E.(2002)Novel Bacillus thuringiensis binaryinsecticidal crystal proteins active on western corn rootworm,Diabroticavirgifera virgifera LeConte.Appl.Environ.Microbiol.68:1137-1145.Emsley,P.,Bernhard Lohkamp,William G.Scott,and Kevin Cowtan.(2010).“Features andDevelopment of Coot.”Acta Cryst D66:486-501.

Englemann,F.,Geraerts,W.P.M.,(1980)The proteases and the proteaseinhibitor in the midgut of Leucophaea maderae.J.Insect Physiol.261:703-710.Fallahi,M.,Crosthwait,J.,Calixte,S.,and Bonen,L.,(2005).“Fate ofMitochondrially Located S19 Ribosomal Protein Genes after Transfer of aFunctional Copy to the Nucleus in Cereals”Mol Gen Genomics 273:76–83.

Fraley,R.T.,Rogers,S.G.,Horsch,R.B.(1986)Genetic transformation inhigher plants.Crit.Rev.Plant Sci.4:1-46.

Frankenhuyzen,K.,(2009)Insecticidal activity of Bacillusthuringiensis crystal proteins.Journal of Invertebrate Pathology,Vol:101(1):1-16.

Gallagher,S.,Winston,S.,Fuller,S.,Hurrell,J.(2008)Immunoblotting andImmunodetection.Current Protocols in immunology 8.10.1–8.10.28.

Gazit,E.,La Rocca,P.,Sansom,M.S.P.,Shai,Y.(1998)The structure andorganization within the membrane of the helices composing the pore-formingdomain of Bacillus thuringiensis delta-endotoxin are consistent with an"umbrella-like"structure of the pore.Proc.Nat.Acad.Sci.USA 95:12289-12294.Ge,A.,Rivers,D.,Milne,R.,Dean,D.H.(1991)Functional domains of Bacillusthuringiensis insecticidal crystal proteins.Refinement of Heliothis virescensand Trichoplusia ni specificity domains on CryIA(c).J.Biol.Chem.266:17954-17958.

Gillikin,J.W.,Bevilacqua,S.,Graham,J.S.(1992)Partial characterizationof digestive tract proteinases from western corn rootworm larvae,Diabroticavirgifera.Arch.Insect Biochem.Physiol.19:285-298.

Gnanasambandam,A.,Anderson,D.J.,Mills,E.,and Brumbley,S.M.(2012).“Heterologous C-terminal Signals Effectively Target Fluorescent FusionProteins to Leaf Peroxisomes in Diverse Plant Species”Journal of PlantPhysiology 169:830–833.

Gomez,I.,Sanchez,J.,Miranda,R.,Bravo,A.,Soberon,M.(2002)Cadherin-likereceptor binding facilitates proteolytic cleavage of helix alpha-1 in domainI and oligomer pre-pore formation of Bacillus thuringiensis Cry1Ab toxin.FEBSLett.513:242-246.

Hahn,J.J.,Eschenlauer,A.C.,Sleyter,U.B.,Somers,D.A.and Srienc,F.,(1999)“Peroxisomes as Sites for Synthesis of Polyhydroxyalkanoates inTransgenic Plants”Biotechnol.Prog.15:1053-1057.

Haider,M.Z.,Knowles,B.H.,Ellar,D.J.(1986)Specificity of Bacillusthuringiensis var.colmeri insecticidalδ-endotoxin is determined bydifferential proteolytic processing of the protoxin by larval gutproteases.Eur.J.Biochem.156:531-540.

Heckel,D.G.,Gahan,L.J.,Baxter,S.W.,Zhao,J-Z.,Shelton,A.M.,Gould,F.,Tabashnik,B.E.(2007)The diversity of Bt resistance genes in species ofLepidoptera.J.Invert.Pathol.95:192-197.

Hepburn,A.G.,White,J.,Pearson,L.,Maunders,M.J.,Clarke,L.E.,Prescott,A.G.Blundy,K.S.(1985)The use of pNJ5000 as an intermediate vector for thegenetic manipulation of Agrobacterium Ti-plasmids.J.Gen.Microbiol.131:2961-2969.

Hoagland,D.R.,Arnon,D.I.(1950)The water-culture method of growingplants without soil.Calif.Agr.Expt.Sta.Circ.347.

Hofte,H.,de Greve,H.,Seurinck,J.,Jansens,S.,Mahillon,J.,Ampe,C.,Vandekerckhove,J.,Vanderbruggen,H.,van Montagu,M.,Zabeau,M.,Vaeck,M.(1986)Structural and functional analysis of a cloned delta endotoxin of Bacillusthuringiensis berliner 1715.Eur.J.Biochem.161:273-280.

Holwerda,B.,Padgett,H.and Rogers,J.(1992).“Proaleurain VacuolarTargeting Is Mediated by Short Contiguous Peptide lnteractions”.The PlantCell 4:307-318.

Honée,G.,Convents,D.,Van Rie,J.,Jansens,S.,Peferoen,M.,Visser,B.(1991)The C-terminal domain of the toxic fragment of a Bacillus thuringiensiscrystal protein determines receptor binding.Mol.Microbiol.5:2799-2806

Hood,E.E.,Love,R.,Lane,J.,Bray,J.,Clough,R.,Pappu,K.,Drees,C.,Hood,K.R.,Yoon,S.,Ahmad,A.,and Howard,J.A.(2007).“Subcellular Targeting is a KeyCondition for High-Level Accumulation of Cellulase Protein in TransgenicMaize Seed”.Plant Biotechnolgy Journal 5:709-719.

Horton,R.M.,Hunt,H.D.,Ho,S.N.,Pullen,J.K.,Pease,L.R.(1989)Engineeringhybrid genes without the use of restriction enzymes:gene splicing by overlapextension.Gene 77:61–68.

Huang,F.,Rogers,L.B.,Rhett,G.H.(2006)Comparative susceptibility ofEuropean corn borer,southwestern corn borer,and sugarcane borer(Lepidoptera:Crambidae)to Cry1Ab protein in a commercial Bacillus thuringiensis cornhybrid.J.Econ.Entomol.99:194-202.

Huang,K-X.,Badger,M.,Haney,K.,Evans,S.L.(2007)Large scale productionof Bacillus thuringiensis PS149B1 insecticidal proteins Cry34Ab1 andCry35Ab1from Pseudomonas fluorescens.Prot.Express.Purific.53:325-330.

Hyunjong,B.,Lee,D-S.and Hwang,I.(2006).“Dual Targeting of Xylanase toChloroplasts and Peroxisomes as a Means to Increase Protein Accumulation inPlant Cells”J.Exp.Bot.57:161-169.

Janmaat,A.F.,Myers,A.H.(2003)Rapid evolution and the cost ofresistance to Bacillus thuringiensis in greenhouse populations of cabbageloopers,Trichoplusia ni.Proc.Royal Soc.London.Ser.B,Biolog.Sci.270:2263-2270.

Janmaat,A.F.,Myers,A.H.(2005)The cost of resistance to Bacillusthuringiensis varies with the host plant of Trichoplusia ni.Proc.RoyalSoc.London.Ser.B,Biolog.Sci.272:1031-1038.

Karlin,S.,Altschul,S.F.(1990)Methods for assessing the statisticalsignificance of molecular sequence features by using general scoringschemes.Proc.Natl.Acad.Sci.USA 87:2264-2268.

Karlin,S.,Altschul,S.F.(1993)Applications and statistics for multiplehigh-scoring segments in molecular sequences.Proc.Natl.Acad.Sci.USA90:5873-5877.

Keller,G.H.,Manak,M.M.(1993)DNA Probes,Background,Applications,Procedures.Stockton Press,New York,NY.

Knight,J.S.,Broadwell,A.H.,Grant,W.N.,Shoemaker,C.B.(2004)A Strategyfor Shuffling Numerous Bacillus thuringiensis Crystal ProteinDomains.J.Econ.Entomol.97:1805-1813.

Koiwa,H.,Shade,R.E.,Zhu-Salzman,K.,D'Urzo,M.P.,Murdock,L.L.,Bressan,R.A.,Hasegawa,P.M.(2000)A plant defensive cystatin(soyacystatin)targetscathepsin L-like digestive cysteine proteinases(DvCALs)in the larval midgutof western corn rootworm Diabrotica virgifera virgifera.FEBS Letters471:67-70.

Larson,S.M.,England,J.L.,Desjarlais,J.R.,Pande,V.S.(2002)Thoroughlysampling sequence space:Large-scale protein design of structuralensembles.Protein Sci.11:2804-2813.

Lee,L.-Y.,Gelvin,S.B.(2008)T-DNA binary vectors and systems.PlantPhysiol.146:325-332.

Li,H.,Olson,M.,Lin,G.,Hey,T.,Tan,S.Y.and Narva,K.(2013).“Bacillusthuringiensis Cry34Ab1/Cry35Ab1 Interactions with Western Corn RootwormMidgut Membrane Binding Sites”.PLoS One 8:e53079.

Linsmaier,E.M.,Skoog,F.(1965)Organic growth factor requirements oftobacco tissue.Physiologia Plant 18:100-127.

Littlefield,J.W.(1964)Selection of hybrids from matings offibroblasts in vitro and their presumed recombinants.Science 145:709-710.

Mahendra Madegowda,Subramaniam Eswaramoorthy,Stephen K.Burley,andSubramanyam Swaminathan(2008).“X-ray crystal structure of the B component ofHemolysin BL from Bacillus cereus”Proteins.2008 May 1；71(2):534–540.

Mano,S.,Nakamori,C.,Hayashi,M.,Kato,A.,Kondo,M.,Nishimura,M.(2002).“Distribution and Characterization of Peroxisomes in Arabdopsis byVisualization with GFP:Dynamic Morphology and Actin-Dependent Movement”.PlantCell Physiol.43:331-341.

McCoy,A.J.,Grosse-Kunstleve,R.W.,Adams,P.D.,Winn,M.D.,Storoni,L.C.,&Read,R.J.(2007).“Phaser crystallographic software.”J.Appl.Cryst.40:658-674.

Meinkoth,J.,Wahl,G.(1984)Hybridization of nucleic acids immobilizedon solid supports.Anal.Biochem.138:267-284.

Metcalf,R.L.(1986)The ecology of insecticides and the chemicalcontrol of insects.pp.251-297.In(Marcos Kogan(ed.))Ecological theory andintegrated pest management practice.John Wiley&Sons,N.Y.362 pp.

Miao,Y.,Yan P.K.,Kim,H.,Hwang,I.,and Jiang,L.(2006).“Localization ofGreen Fluorescent Protein Fusions with the Seven Arabidopsis Vacuolar SortingReceptors to Pre-vacuolar Compartments in Tobacco BY-2 Cells”.PlantPhysiol.142:945-962.

Moellenbeck,D.J.,Peters,M.L.,Bing,J.W.,Rouse,J.R.,Higgins,L.S.,Sims,L.,Nevshemal,T.,Marshall,L.,Ellis,R.T.,Bystrak,P.G.,Lang,B.A.,Stewart,J.L.,Kouba,K.,Sondag,V.,Gustafson,V.,Nour,K.,Xu,D.,Swenson,J.,Zhang,J.,Czapla,T.,Schwab,G.,Jayne,S.,Stockhoff,B.A.,Narva,K.,Schnepf,H.E.,Stelman,S.J.,Poutre,C.,Koziel,M.,Duck,N.(2001)Insecticidal proteins from Bacillus thuringiensisprotect corn from corn rootworms.Nat.Biotech.19:668-672.

Mueller,Marcus,Ulla Grauschopf,Timm Maier,Rudi Glockshuber and NenadBan(2009).“The structure of a cytolytic a-helical toxin pore reveals itsassembly mechanism”.Nature Vol 459:4 June 2009:726.

Murashige,T.and Skoog,F.(1962).A revised medium for rapid growth andbioassays with tobacco tissue cultures.Physiol.Plant 15:473–497.

Murshudov,G.N.,A.A.Vagin and E.J.Dodson,(1997)."Refinement ofMacromolecular Structures by the Maximum-Likelihood method."Acta Cryst.D53:240-255.

Myers,E.,Miller,W.(1988)Optimal alignments in linear space.CABIOS4:11-17.

Naimov,S.,Weemen-Hendriks,M.,Dukiandjiev,S.,de Maagd,R.A.(2001)Bacillus thuringiensis delta-endotoxin Cry1 hybrid proteins with increasedactivity against the Colorado potato beetle.Appl.Environ.Microbiol.11:5328-5330.

Needleman,S.B.,Wunsch,C.D.(1970)A general method applicable to thesearch for similarities in the amino acid sequence of twoproteins.J.Mol.Biol.48:443-453.

Nora Eifler,Michael Vetsch,Marco Gregorini,Philippe Ringler,MohamedChami,Ansgar Philippsen,Andrea Fritz,Shirley A Muller,RudiGlockshuber,Andreas Engel,and Ulla Grauschopf(2006).“Cytotoxin ClyA fromEscherichia coli assembles to a 13-meric pore independent of its redox-state”The EMBO Journal25:2652–2661

Nunez-Valdez,M.-E.,Sanchez,J.,Lina,L.,Guereca,L.,Bravo,A.(2001)Structural and functional studies of alpha-helix 5 region from Bacillusthuringiensis Cry1Ab delta-endotoxin.Biochim.Biophys.Acta,Prot.Struc.Molec.Enzymol.1546:122-131.

Ochoa-Campuzano,C.,Real,M.D.,Martinez-Ramirez,A.C.,Bravo,A.,Rausell,

C.(2007)An ADAM metalloprotease is a Cry3Aa Bacillus thuringiensistoxin receptor.Biochem.Biophys.Res.Commun.362:437-442.

Oleson JD1,Park YL,Nowatzki TM,Tollefson JJ.(2005).“Node-injury scaleto evaluate root injury by corn rootworms(Coleoptera:Chrysomelidae)”J EconEntomol.2005 Feb；98(1):1-8.

Otwinowski,Z.and W.Minor,"Processing of X-ray Diffraction DataCollected in Oscillation Mode",Methods in Enzymology,Volume 276:Macromolecular Crystallography,part A,p.307-326,1997,C.W.Carter,Jr.&R.M.Sweet,Eds.,Academic Press(New York).

Pigott,C.R.,Ellar,D.J.(2007)Role of receptors in Bacillusthuringiensis crystal toxin activity.Microbiol.Molec.Biol.Rev.71:255-281.

Prichard,R.K.et al.“The problem of anthelmintic resistance innematodes,”Austr.Vet.J.56:239-251；Coles,G.C.“Anthelmintic resistance insheep,”In Veterinary Clinics of North America:Food Animal Practice,Vol 2:423-432[Herd,R.P.,eds.]W.B.Saunders,New York

Rang,C.,Vachon,V.,de Maagd,R.A.,Villalon,M.,Schwartz,J.-L.,Bosch,D.,Frutos,R.,Laprade R.(1999)Interaction between functional domains of Bacillusthuringiensis insecticidal crystal proteins.Appl.Environ.Microbiol.65:2918-2925.

Rokov-Plavec,Dulic,M.,Duchene,A-M.,Weygand-Durasevic,I.(2008).“DualTargeting of Organellar Seryl-tRNA Synthetase to Maize Mitochondria andChloroplasts”Plant Cell Rep 27:1157-1168.

Sambrook,J.,Fritsch,E.F.,Maniatis,T.(1989)Molecular Cloning:ALaboratory Manual(2nd ed.,Cold Spring Harbor Laboratory Press,Plainview,N.Y.)

Schenk,R.U.,Hildebrandt,A.C.(1972)Medium and techniques for inductionand growth of monocotyledonous and dicotyledonous plant cellcultures.Can.J.Bot.50:199-204

Schnepf,H.E.,Tomczak,K.,Ortega,J.P.,Whiteley,H.R.(1990)Specificity-determining regions of a Lepidopteran-specific insecticidal protein producedby Bacillus thuringiensis.J.Biol.Chem.265:20923-20930.

Soberon,M.,Pardo-Lopez,L.,Lopez,I.,Gomez,I.,Tabashnik,B.E.,Bravo,A.(2007)Engineering modified Bt toxins to counter insect resistance.Science318:1640-1642.

Squires,C.H.,Retallack,D.M.,Chew,L.C.,Ramseier,T.M.,Schneider,J.C.,Talbot,H.W.(2004)Heterologous protein production in P.fluorescens.BioprocessIntern.2:54-59.

Stemmer,W.P.C.(1994a)DNA shuffling by random fragmentation andreassembly:in vitro recombination for molecularevolution.Proc.Natl.Acad.Sci.USA 91:10747-10751

Stemmer,W.P.C.(1994b)Rapid evolution of a protein in vitro by DNAshuffling.Nature 370:389-391.

Stemmer,W.P.C.(1995)Searching sequence space.Bio/Technology13:549-553.

Stewart,L.(2007)Gene synthesis for protein production.Encylopedia ofLife Sciences.John Wiley and Sons,Ltd.

Stewart,L.,Burgin,A.B.,(2005)Whole gene synthesis:a gene-o-maticfuture.Frontiers in Drug Design and Discovery 1:297-341.

Suggs,S.V.,Miyake,T.,Kawashime,E.H.,Johnson,M.J.,Itakura,K.,R.B.Wallace,R.B.(1981)ICN-UCLA Symposium.Dev.Biol.Using Purified Genes,

D.D.Brown(ed.),Academic Press,New York,23:683-69

Tabashnik,B.E.,Finson,N.,Groeters,F.R.,Moar,W.J.,Johnson,M.W.,Luo,K.,Adang,M.J.(1994)Reversal of resistance to Bacillus thuringiensis in Plutellaxylostella.Proc.Nat.Acad.Sci.USA 91:4120-4124.

Tabashnik,B.E.,Gassmann,A.J.,Crowder,D.W.,Carriere,T.(2008)Insectresistance to Bt crops:evidence versus theory.Nat.Biotech.26:199-202.

Taggart,R.T.,Samloff,I.M.(1983)Stable antibody-producing murinehybridomas.Science 219:1228-1230.

Terry R.Wright,Guomin Shana,Terence A.Walsh,Justin M.Lira,Cory Cui,Ping Song,Meibao Zhuang,Nicole L.Arnold,Gaofeng Lin,Kerrm Yau,Sean M.Russell,Robert M.Cicchillo,Mark A.Peterson,David M.Simpson,Ning Zhou,JayakumarPonsamuel,and Zhanyuan Zhang.(2010)“Robust crop resistance to broadleaf andgrass herbicides provided by aryloxyalkanoate dioxygenase transgenes”.20240–20245|PNAS|November 23,2010|vol.107|no.47.

Thie,N.M.R.,Houseman J.G.(1990)Identification of cathepsin B,D and Hin the larval midgut of Colorado potato beetle,Leptinotarsa decemlineata say(Coleoptera:Chrysomelidae)Insect Biochem.20:313-318.

Thompson,J.D.,Higgins,D.G.,Gibson,T.J.(1994)CLUSTAL W:improving thesensitivity of progressive multiple sequence alignment through sequenceweighting,position-specific gap penalties and weight matrix choice.Nucl.AcidsRes.22:4673-4680.

Tijssen,P.(1993)Laboratory Techniques in Biochemistry and MolecularBiology Hybridization with Nucleic Acid Probes,Part I,Chapter 2.P.C.van derVliet(ed.),(Elsevier,N.Y.)

Tzokov,S.B.,Wyborn,N.R.,Stillman,T.J.,Jamieson,S.,Czudnochowski,N.,Artymuik,P.J.,Green,J.,Bullough,P.A.(2006).“Sturcture of the Hemolysin E(HlyE,ClyA and SheA)Channel in Its Mebrane-bound Form”.J.Biol.Chem.2006,281:23042-23049.

Varagona,M.,and Raikhel,N.(1994).“The Basic Domain in the bZIPRegulatory Protein Opaque2 Serves Two Independent Functions:DNA Binding andNuclear Localization”.The Plant Journal 5:207-214.

Varshavsky,A.(1997)The N-end rule pathway of proteindegradation.Genes to Cells 2:13-28.

Vaughn,T.,Cavato,T.,Brar,G.,Coombe,T.,DeGooyer,T.,Ford,S.,Groth,M.,Howe,A.,Johnson,S.,Kolacz,K.,Pilcher,C.,Prucell,J.,Romano,C.,English,L.,Pershing,J.(2005)A method of controlling corn rootworm feeding using aBacillus thuringiensis protein expressed in transgenic maize.Crop.Sci.45:931–938.

Walters,F.S.,Slatin,S.L.,Kulesza,C.A.,English,L.H.(1993)Ion channelactivity of N-terminal fragments from CryIA(c)delta-endotoxin.Biochem.Biophys.Res.Commun.196:921-926.

Walters,F.S.,Stacy,C.M.,Lee,M.K.,Palekar,N.,Chen,J.S.(2008)Anengineered chymotrypsin/cathepsin G site in domain I renders Bacillusthuringiensis Cry3A active against western corn rootwormlarvae.Appl.Environ.Microbiol.74:367-374.

Wehrmann,A.,Van Vliet,A.,Opsomer,C.,Botterman,J.,Schulz,A.(1996)Thesimilarities of bar and pat gene products make them equally applicable forplant engineers.Nat.Biotechnol.14:1274-1278.

Weigel,D.,Glazebrook,J.(eds.)(2002)Arabidopsis:A LaboratoryManual.Cold Spring Harbor Press,Cold Spring Harbor,NY,354 pages.

Wei,J-Z.,Hale,K.,Carta,L.,Platzer,E.,Wong,C.,Fang,S-C.,Aroian,R.(2003).“Bacillus thuringiensis crystal proteins that target nematodes”.PNAS100:2760-2765.

White,J.A.,and Scandalios,J.G.(1989).“Deletion Analysis of the MaizeMitochondrial Superoxide Dismutase Transit Peptide”PNAS 86:3534-3538.

Winn,M.D.et al.(2011)."Overview of the CCP4 suite and currentdevelopments."Acta.Cryst.D67:235-242.

Witkowski,J.F.,Wedberg,J.L.,Steffey,K.L.,Sloderbeck,P.E.,Siegfried,B.D.,Rice,M.E.,Pilcher,C.D.,Onstad,D.W.,Mason,C.E.,Lewis,L.C.,Landis,D.

A.,Keaster,A.J.,Huang,F.,Higgins,R.A.,Haas,M.J.,Gray,M.E.,Giles,K.L.,Foster,J.E.,Davis,P.M.,Calvin,D.D.,Buschman,L.L.,Bolin,P.C.,Barry,B.D.,Andow,D.A.,Alstad,D.N.(2002)Bt corn and European Corn Borer(Ostlie,K.R.,Hutchison,W.D.,Hellmich,R.L.(eds)).University of Minnesota Extension Service.Publ.WW-07055.

Wolfson,J.L.,Murdock,L.L.(1990)Diversity in digestive proteinaseactivity among insects.J.Chem.Ecol.16:1089-1102.

Worley,C.K.,Ling,R.,Callis,J.(1998)Engineering in vivo instability offirefly luciferase and Escherichia coliβ-glucuronidase in higher plants usingrecognition elements from the ubiquitin pathway.Plant Molec.Biol.37:337-347.

序列表

<110> Dow AgroSciences LLC

Hey, Tim

Xu, Xiaoping

Glancy, Todd

AlAbed, Diaa

Worden, Sarah

Wang, Nick

Ausmus, Carla

Hung, Shao-Ching

<120> 工程化的CRY6A杀虫蛋白

<130> 77095

<160> 144

<170> PatentIn 3.5版

<210> 1

<211> 1428

<212> DNA

<213> 苏云金芽孢杆菌

<400> 1

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa tgtcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattaa 1428

<210> 2

<211> 475

<212> PRT

<213> 苏云金芽孢杆菌

<400> 2

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

465 470 475

<210> 3

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 3

atgatcattg actccaagac caccctgccg cgtcacagcc tgatccacac catcaagctg 60

aactccaaca agaagtacgg acctggcgac atgaccaacg gcaaccagtt catcatctcc 120

aagcaagagt gggccaccat cggagcgtac atccagaccg gcctgggcct gccggtgaac 180

gagcagcagc tgaggactca cgtgaacctg tcccaagaca tcagcatccc gtccgacttc 240

tcccagctgt acgacgtgta ttgcagcgac aagacctccg ccgagtggtg gaacaagaac 300

ctgtacccgc tgatcatcaa gagcgccaac gacatcgcct cctacggctt caaggtggcg 360

ggcgacccga gcatcaagaa ggacggctac ttcaagaagc tccaagacga gctggacaac 420

atcgtggaca acaactccga cgacgacgcc atcgccaagg cgatcaagga cttcaaggcg 480

aggtgcggca tcctgatcaa ggaggcgaag cagtacgagg aggcggcgaa gaacatcgtg 540

acctccctgg accagttcct gcacggcgac cagaagaagc tggagggcgt gatcaacatc 600

cagaagaggc tgaaggaggt gcagaccgcc ctgaatcaag cccacggcga gagcagcccg 660

gcgcacaagg agctgctgga gaaggtgaag aacctcaaga ccaccctgga gaggaccatc 720

aaggccgagc aagacctgga gaagaaggtg gagtactcct tcctgctggg tccgctgctg 780

ggcttcgtgg tgtacgagat cctggagaac accgccgtgc agcacatcaa gaatcagatc 840

gacgagatca agaagcagct ggactccgcc cagcacgacc tggaccgcga cgtgaagatc 900

atcgggatgc tgaacagcat caacaccgac atcgacaacc tgtacagcca aggccaagag 960

gccatcaagg tgttccagaa gctccaaggc atctgggcga ccatcggcgc gcagatcgag 1020

aacctgagga ccaccagcct ccaagaggtc caagacagcg acgacgccga cgagatccag 1080

atcgagctgg aggacgccag cgacgcgtgg ctggtggtgg cccaagaggc gagggacttc 1140

accctgaacg cgtacagcac caactcccgt cagaacctgc cgatcaacgt gatcagcgac 1200

agctgcaact gctccaccac caacatgacc agcaaccagt actccaaccc gaccaccaac 1260

atgaccagca accagtacat gatctcccac gagtacacca gcctgccgaa caacttcatg 1320

ctgtcccgca actctaacct ggagtacaag tgcccggaga acaacttcat gatctactgg 1380

tacaacaact ccgactggta caacaactca gactggtaca acaactga 1428

<210> 4

<400> 4

000

<210> 5

<211> 529

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 5

Met Ala Ile Gly Ser Leu Ile Ala Ser Thr Phe Ala Arg Ser Ser His

1 5 10 15

Ala Leu Pro Ala Ala Ala Ala Ser Ala Ile Ser Gln Ala Pro Arg Ser

20 25 30

Gln His Thr Ala Ser Pro Leu Leu Ser Gly Leu Gly Ala Ala Ala Arg

35 40 45

Ala Phe Ser Ser Arg Ala Leu Ile Ile Asp Ser Lys Thr Thr Leu Pro

50 55 60

Arg His Ser Leu Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr

65 70 75 80

Gly Pro Gly Asp Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln

85 90 95

Glu Trp Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro

100 105 110

Val Asn Glu Gln Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp Ile

115 120 125

Ser Ile Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp

130 135 140

Lys Thr Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile

145 150 155 160

Lys Ser Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp

165 170 175

Pro Ser Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu

180 185 190

Asp Asn Ile Val Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala

195 200 205

Ile Lys Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys

210 215 220

Gln Tyr Glu Glu Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln Phe

225 230 235 240

Leu His Gly Asp Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln Lys

245 250 255

Arg Leu Lys Glu Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu Ser

260 265 270

Ser Pro Ala His Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys Thr

275 280 285

Thr Leu Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys Val

290 295 300

Glu Tyr Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr Glu

305 310 315 320

Ile Leu Glu Asn Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp Glu

325 330 335

Ile Lys Lys Gln Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp Val

340 345 350

Lys Ile Ile Gly Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu

355 360 365

Tyr Ser Gln Gly Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln Gly

370 375 380

Ile Trp Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser

385 390 395 400

Leu Gln Glu Val Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu

405 410 415

Leu Glu Asp Ala Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala Arg

420 425 430

Asp Phe Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro

435 440 445

Ile Asn Val Ile Ser Asp Ser Cys Asn Cys Ser Thr Thr Asn Met Thr

450 455 460

Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr

465 470 475 480

Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser

485 490 495

Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile

500 505 510

Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

515 520 525

Asn

<210> 6

<211> 1518

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 6

atggccctta ggaccttagc gagcaagaag gtgctgagct tcccgttcgg cggagcgggc 60

agaccgttag cagcagcggc ctcagctagg ggcatcattg actccaagac caccctgccg 120

cgtcacagcc tgatccacac catcaagctg aactccaaca agaagtacgg acctggcgac 180

atgaccaacg gcaaccagtt catcatctcc aagcaagagt gggccaccat cggagcgtac 240

atccagaccg gcctgggcct gccggtgaac gagcagcagc tgaggactca cgtgaacctg 300

tcccaagaca tcagcatccc gtccgacttc tcccagctgt acgacgtgta ttgcagcgac 360

aagacctccg ccgagtggtg gaacaagaac ctgtacccgc tgatcatcaa gagcgccaac 420

gacatcgcct cctacggctt caaggtggcg ggcgacccga gcatcaagaa ggacggctac 480

ttcaagaagc tccaagacga gctggacaac atcgtggaca acaactccga cgacgacgcc 540

atcgccaagg cgatcaagga cttcaaggcg aggtgcggca tcctgatcaa ggaggcgaag 600

cagtacgagg aggcggcgaa gaacatcgtg acctccctgg accagttcct gcacggcgac 660

cagaagaagc tggagggcgt gatcaacatc cagaagaggc tgaaggaggt gcagaccgcc 720

ctgaatcaag cccacggcga gagcagcccg gcgcacaagg agctgctgga gaaggtgaag 780

aacctcaaga ccaccctgga gaggaccatc aaggccgagc aagacctgga gaagaaggtg 840

gagtactcct tcctgctggg tccgctgctg ggcttcgtgg tgtacgagat cctggagaac 900

accgccgtgc agcacatcaa gaatcagatc gacgagatca agaagcagct ggactccgcc 960

cagcacgacc tggaccgcga cgtgaagatc atcgggatgc tgaacagcat caacaccgac 1020

atcgacaacc tgtacagcca aggccaagag gccatcaagg tgttccagaa gctccaaggc 1080

atctgggcga ccatcggcgc gcagatcgag aacctgagga ccaccagcct ccaagaggtc 1140

caagacagcg acgacgccga cgagatccag atcgagctgg aggacgccag cgacgcgtgg 1200

ctggtggtgg cccaagaggc gagggacttc accctgaacg cgtacagcac caactcccgt 1260

cagaacctgc cgatcaacgt gatcagcgac agctgcaact gctccaccac caacatgacc 1320

agcaaccagt actccaaccc gaccaccaac atgaccagca accagtacat gatctcccac 1380

gagtacacca gcctgccgaa caacttcatg ctgtcccgca actctaacct ggagtacaag 1440

tgcccggaga acaacttcat gatctactgg tacaacaact ccgactggta caacaactca 1500

gactggtaca acaactga 1518

<210> 7

<211> 505

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 7

Met Ala Leu Arg Thr Leu Ala Ser Lys Lys Val Leu Ser Phe Pro Phe

1 5 10 15

Gly Gly Ala Gly Arg Pro Leu Ala Ala Ala Ala Ser Ala Arg Gly Ile

20 25 30

Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His Thr Ile

35 40 45

Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr Asn Gly

50 55 60

Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly Ala Tyr

65 70 75 80

Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu Arg Thr

85 90 95

His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe Ser Gln

100 105 110

Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp Trp Asn

115 120 125

Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile Ala Ser

130 135 140

Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp Gly Tyr

145 150 155 160

Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn Asn Ser

165 170 175

Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala Arg Cys

180 185 190

Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala Lys Asn

195 200 205

Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys Lys Leu

210 215 220

Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln Thr Ala

225 230 235 240

Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu Leu Leu

245 250 255

Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile Lys Ala

260 265 270

Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu Gly Pro

275 280 285

Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala Val Gln

290 295 300

His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp Ser Ala

305 310 315 320

Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu Asn Ser

325 330 335

Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu Ala Ile

340 345 350

Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly Ala Gln

355 360 365

Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp Ser Asp

370 375 380

Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp Ala Trp

385 390 395 400

Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala Tyr Ser

405 410 415

Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp Ser Cys

420 425 430

Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn Pro Thr

435 440 445

Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser

450 455 460

Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys

465 470 475 480

Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp

485 490 495

Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

500 505

<210> 8

<211> 1665

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 8

atgctgacct gcggaaggtt catcagctcc agcgcggcca ccagcaccgc ctccttcttc 60

ccgtttagga ccctgacaag gagcctggtg aggagaccgg ccccgcacct gctcagctcc 120

gcgtcagctg ccgccgccac agcggtggag ctggacacca acggagacgg ctcagcgggc 180

ggcggagccg gcgtggtgag gcctcagtgg aaggcggcaa tcgacttcaa gtggatcagg 240

atcattgact ccaagaccac cctgccgcgt cacagcctga tccacaccat caagctgaac 300

tccaacaaga agtacggacc tggcgacatg accaacggca accagttcat catctccaag 360

caagagtggg ccaccatcgg agcgtacatc cagaccggcc tgggcctgcc ggtgaacgag 420

cagcagctga ggactcacgt gaacctgtcc caagacatca gcatcccgtc cgacttctcc 480

cagctgtacg acgtgtattg cagcgacaag acctccgccg agtggtggaa caagaacctg 540

tacccgctga tcatcaagag cgccaacgac atcgcctcct acggcttcaa ggtggcgggc 600

gacccgagca tcaagaagga cggctacttc aagaagctcc aagacgagct ggacaacatc 660

gtggacaaca actccgacga cgacgccatc gccaaggcga tcaaggactt caaggcgagg 720

tgcggcatcc tgatcaagga ggcgaagcag tacgaggagg cggcgaagaa catcgtgacc 780

tccctggacc agttcctgca cggcgaccag aagaagctgg agggcgtgat caacatccag 840

aagaggctga aggaggtgca gaccgccctg aatcaagccc acggcgagag cagcccggcg 900

cacaaggagc tgctggagaa ggtgaagaac ctcaagacca ccctggagag gaccatcaag 960

gccgagcaag acctggagaa gaaggtggag tactccttcc tgctgggtcc gctgctgggc 1020

ttcgtggtgt acgagatcct ggagaacacc gccgtgcagc acatcaagaa tcagatcgac 1080

gagatcaaga agcagctgga ctccgcccag cacgacctgg accgcgacgt gaagatcatc 1140

gggatgctga acagcatcaa caccgacatc gacaacctgt acagccaagg ccaagaggcc 1200

atcaaggtgt tccagaagct ccaaggcatc tgggcgacca tcggcgcgca gatcgagaac 1260

ctgaggacca ccagcctcca agaggtccaa gacagcgacg acgccgacga gatccagatc 1320

gagctggagg acgccagcga cgcgtggctg gtggtggccc aagaggcgag ggacttcacc 1380

ctgaacgcgt acagcaccaa ctcccgtcag aacctgccga tcaacgtgat cagcgacagc 1440

tgcaactgct ccaccaccaa catgaccagc aaccagtact ccaacccgac caccaacatg 1500

accagcaacc agtacatgat ctcccacgag tacaccagcc tgccgaacaa cttcatgctg 1560

tcccgcaact ctaacctgga gtacaagtgc ccggagaaca acttcatgat ctactggtac 1620

aacaactccg actggtacaa caactcagac tggtacaaca actga 1665

<210> 9

<211> 554

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 9

Met Leu Thr Cys Gly Arg Phe Ile Ser Ser Ser Ala Ala Thr Ser Thr

1 5 10 15

Ala Ser Phe Phe Pro Phe Arg Thr Leu Thr Arg Ser Leu Val Arg Arg

20 25 30

Pro Ala Pro His Leu Leu Ser Ser Ala Ser Ala Ala Ala Ala Thr Ala

35 40 45

Val Glu Leu Asp Thr Asn Gly Asp Gly Ser Ala Gly Gly Gly Ala Gly

50 55 60

Val Val Arg Pro Gln Trp Lys Ala Ala Ile Asp Phe Lys Trp Ile Arg

65 70 75 80

Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His Thr

85 90 95

Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr Asn

100 105 110

Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly Ala

115 120 125

Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu Arg

130 135 140

Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe Ser

145 150 155 160

Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp Trp

165 170 175

Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile Ala

180 185 190

Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp Gly

195 200 205

Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn Asn

210 215 220

Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala Arg

225 230 235 240

Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala Lys

245 250 255

Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys Lys

260 265 270

Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln Thr

275 280 285

Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu Leu

290 295 300

Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile Lys

305 310 315 320

Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu Gly

325 330 335

Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala Val

340 345 350

Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp Ser

355 360 365

Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu Asn

370 375 380

Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu Ala

385 390 395 400

Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly Ala

405 410 415

Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp Ser

420 425 430

Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp Ala

435 440 445

Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala Tyr

450 455 460

Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp Ser

465 470 475 480

Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn Pro

485 490 495

Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr

500 505 510

Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr

515 520 525

Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp

530 535 540

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

545 550

<210> 10

<211> 1446

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 10

atgatcattg actccaagac caccctgccg cgtcacagcc tgatccacac catcaagctg 60

aactccaaca agaagtacgg acctggcgac atgaccaacg gcaaccagtt catcatctcc 120

aagcaagagt gggccaccat cggagcgtac atccagaccg gcctgggcct gccggtgaac 180

gagcagcagc tgaggactca cgtgaacctg tcccaagaca tcagcatccc gtccgacttc 240

tcccagctgt acgacgtgta ttgcagcgac aagacctccg ccgagtggtg gaacaagaac 300

ctgtacccgc tgatcatcaa gagcgccaac gacatcgcct cctacggctt caaggtggcg 360

ggcgacccga gcatcaagaa ggacggctac ttcaagaagc tccaagacga gctggacaac 420

atcgtggaca acaactccga cgacgacgcc atcgccaagg cgatcaagga cttcaaggcg 480

aggtgcggca tcctgatcaa ggaggcgaag cagtacgagg aggcggcgaa gaacatcgtg 540

acctccctgg accagttcct gcacggcgac cagaagaagc tggagggcgt gatcaacatc 600

cagaagaggc tgaaggaggt gcagaccgcc ctgaatcaag cccacggcga gagcagcccg 660

gcgcacaagg agctgctgga gaaggtgaag aacctcaaga ccaccctgga gaggaccatc 720

aaggccgagc aagacctgga gaagaaggtg gagtactcct tcctgctggg tccgctgctg 780

ggcttcgtgg tgtacgagat cctggagaac accgccgtgc agcacatcaa gaatcagatc 840

gacgagatca agaagcagct ggactccgcc cagcacgacc tggaccgcga cgtgaagatc 900

atcgggatgc tgaacagcat caacaccgac atcgacaacc tgtacagcca aggccaagag 960

gccatcaagg tgttccagaa gctccaaggc atctgggcga ccatcggcgc gcagatcgag 1020

aacctgagga ccaccagcct ccaagaggtc caagacagcg acgacgccga cgagatccag 1080

atcgagctgg aggacgccag cgacgcgtgg ctggtggtgg cccaagaggc gagggacttc 1140

accctgaacg cgtacagcac caactcccgt cagaacctgc cgatcaacgt gatcagcgac 1200

agctgcaact gctccaccac caacatgacc agcaaccagt actccaaccc gaccaccaac 1260

atgaccagca accagtacat gatctcccac gagtacacca gcctgccgaa caacttcatg 1320

ctgtcccgca actctaacct ggagtacaag tgcccggaga acaacttcat gatctactgg 1380

tacaacaact ccgactggta caacaactca gactggtaca acaacagggc agtggcgagg 1440

ctgtga 1446

<210> 11

<211> 481

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 11

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Arg Ala Val Ala Arg

465 470 475 480

Leu

<210> 12

<211> 1464

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 12

atgatcattg actccaagac caccctgccg cgtcacagcc tgatccacac catcaagctg 60

aactccaaca agaagtacgg acctggcgac atgaccaacg gcaaccagtt catcatctcc 120

aagcaagagt gggccaccat cggagcgtac atccagaccg gcctgggcct gccggtgaac 180

gagcagcagc tgaggactca cgtgaacctg tcccaagaca tcagcatccc gtccgacttc 240

tcccagctgt acgacgtgta ttgcagcgac aagacctccg ccgagtggtg gaacaagaac 300

ctgtacccgc tgatcatcaa gagcgccaac gacatcgcct cctacggctt caaggtggcg 360

ggcgacccga gcatcaagaa ggacggctac ttcaagaagc tccaagacga gctggacaac 420

atcgtggaca acaactccga cgacgacgcc atcgccaagg cgatcaagga cttcaaggcg 480

aggtgcggca tcctgatcaa ggaggcgaag cagtacgagg aggcggcgaa gaacatcgtg 540

acctccctgg accagttcct gcacggcgac cagaagaagc tggagggcgt gatcaacatc 600

cagaagaggc tgaaggaggt gcagaccgcc ctgaatcaag cccacggcga gagcagcccg 660

gcgcacaagg agctgctgga gaaggtgaag aacctcaaga ccaccctgga gaggaccatc 720

aaggccgagc aagacctgga gaagaaggtg gagtactcct tcctgctggg tccgctgctg 780

ggcttcgtgg tgtacgagat cctggagaac accgccgtgc agcacatcaa gaatcagatc 840

gacgagatca agaagcagct ggactccgcc cagcacgacc tggaccgcga cgtgaagatc 900

atcgggatgc tgaacagcat caacaccgac atcgacaacc tgtacagcca aggccaagag 960

gccatcaagg tgttccagaa gctccaaggc atctgggcga ccatcggcgc gcagatcgag 1020

aacctgagga ccaccagcct ccaagaggtc caagacagcg acgacgccga cgagatccag 1080

atcgagctgg aggacgccag cgacgcgtgg ctggtggtgg cccaagaggc gagggacttc 1140

accctgaacg cgtacagcac caactcccgt cagaacctgc cgatcaacgt gatcagcgac 1200

agctgcaact gctccaccac caacatgacc agcaaccagt actccaaccc gaccaccaac 1260

atgaccagca accagtacat gatctcccac gagtacacca gcctgccgaa caacttcatg 1320

ctgtcccgca actctaacct ggagtacaag tgcccggaga acaacttcat gatctactgg 1380

tacaacaact ccgactggta caacaactca gactggtaca acaacgacct gaaggctctg 1440

gagctgccag tgagcaagct gtga 1464

<210> 13

<211> 487

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 13

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Asp Leu Lys Ala Leu

465 470 475 480

Glu Leu Pro Val Ser Lys Leu

485

<210> 14

<211> 1575

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 14

atgccgaccg acatggagct gtccccgtcc aacgtggcga ggcacagact ggcagttctg 60

gcggcgcacc tgtcagccgc gagcctggag cctccggtga tggcgagcag cctggaggcc 120

cactgcgtta gcgctcagac aatgggcggc atcattgact ccaagaccac cctgccgcgt 180

cacagcctga tccacaccat caagctgaac tccaacaaga agtacggacc tggcgacatg 240

accaacggca accagttcat catctccaag caagagtggg ccaccatcgg agcgtacatc 300

cagaccggcc tgggcctgcc ggtgaacgag cagcagctga ggactcacgt gaacctgtcc 360

caagacatca gcatcccgtc cgacttctcc cagctgtacg acgtgtattg cagcgacaag 420

acctccgccg agtggtggaa caagaacctg tacccgctga tcatcaagag cgccaacgac 480

atcgcctcct acggcttcaa ggtggcgggc gacccgagca tcaagaagga cggctacttc 540

aagaagctcc aagacgagct ggacaacatc gtggacaaca actccgacga cgacgccatc 600

gccaaggcga tcaaggactt caaggcgagg tgcggcatcc tgatcaagga ggcgaagcag 660

tacgaggagg cggcgaagaa catcgtgacc tccctggacc agttcctgca cggcgaccag 720

aagaagctgg agggcgtgat caacatccag aagaggctga aggaggtgca gaccgccctg 780

aatcaagccc acggcgagag cagcccggcg cacaaggagc tgctggagaa ggtgaagaac 840

ctcaagacca ccctggagag gaccatcaag gccgagcaag acctggagaa gaaggtggag 900

tactccttcc tgctgggtcc gctgctgggc ttcgtggtgt acgagatcct ggagaacacc 960

gccgtgcagc acatcaagaa tcagatcgac gagatcaaga agcagctgga ctccgcccag 1020

cacgacctgg accgcgacgt gaagatcatc gggatgctga acagcatcaa caccgacatc 1080

gacaacctgt acagccaagg ccaagaggcc atcaaggtgt tccagaagct ccaaggcatc 1140

tgggcgacca tcggcgcgca gatcgagaac ctgaggacca ccagcctcca agaggtccaa 1200

gacagcgacg acgccgacga gatccagatc gagctggagg acgccagcga cgcgtggctg 1260

gtggtggccc aagaggcgag ggacttcacc ctgaacgcgt acagcaccaa ctcccgtcag 1320

aacctgccga tcaacgtgat cagcgacagc tgcaactgct ccaccaccaa catgaccagc 1380

aaccagtact ccaacccgac caccaacatg accagcaacc agtacatgat ctcccacgag 1440

tacaccagcc tgccgaacaa cttcatgctg tcccgcaact ctaacctgga gtacaagtgc 1500

ccggagaaca acttcatgat ctactggtac aacaactccg actggtacaa caactcagac 1560

tggtacaaca actga 1575

<210> 15

<211> 524

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 15

Met Pro Thr Asp Met Glu Leu Ser Pro Ser Asn Val Ala Arg His Arg

1 5 10 15

Leu Ala Val Leu Ala Ala His Leu Ser Ala Ala Ser Leu Glu Pro Pro

20 25 30

Val Met Ala Ser Ser Leu Glu Ala His Cys Val Ser Ala Gln Thr Met

35 40 45

Gly Gly Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile

50 55 60

His Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met

65 70 75 80

Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile

85 90 95

Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln

100 105 110

Leu Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp

115 120 125

Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu

130 135 140

Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp

145 150 155 160

Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys

165 170 175

Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp

180 185 190

Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys

195 200 205

Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala

210 215 220

Ala Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln

225 230 235 240

Lys Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val

245 250 255

Gln Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys

260 265 270

Glu Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr

275 280 285

Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu

290 295 300

Leu Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr

305 310 315 320

Ala Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu

325 330 335

Asp Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met

340 345 350

Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln

355 360 365

Glu Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile

370 375 380

Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln

385 390 395 400

Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser

405 410 415

Asp Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn

420 425 430

Ala Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser

435 440 445

Asp Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser

450 455 460

Asn Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu

465 470 475 480

Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu

485 490 495

Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn

500 505 510

Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

515 520

<210> 16

<211> 1554

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 16

atggctcacg ctagggtgct gttgcttgcg ctggcggtgc tggcaacagc agccgtggcg 60

gtggcgtcaa gctccagctt cgcggacagc aacccgatta ggcctgtgac cgacagggcc 120

gcctcaacca tcattgactc caagaccacc ctgccgcgtc acagcctgat ccacaccatc 180

aagctgaact ccaacaagaa gtacggacct ggcgacatga ccaacggcaa ccagttcatc 240

atctccaagc aagagtgggc caccatcgga gcgtacatcc agaccggcct gggcctgccg 300

gtgaacgagc agcagctgag gactcacgtg aacctgtccc aagacatcag catcccgtcc 360

gacttctccc agctgtacga cgtgtattgc agcgacaaga cctccgccga gtggtggaac 420

aagaacctgt acccgctgat catcaagagc gccaacgaca tcgcctccta cggcttcaag 480

gtggcgggcg acccgagcat caagaaggac ggctacttca agaagctcca agacgagctg 540

gacaacatcg tggacaacaa ctccgacgac gacgccatcg ccaaggcgat caaggacttc 600

aaggcgaggt gcggcatcct gatcaaggag gcgaagcagt acgaggaggc ggcgaagaac 660

atcgtgacct ccctggacca gttcctgcac ggcgaccaga agaagctgga gggcgtgatc 720

aacatccaga agaggctgaa ggaggtgcag accgccctga atcaagccca cggcgagagc 780

agcccggcgc acaaggagct gctggagaag gtgaagaacc tcaagaccac cctggagagg 840

accatcaagg ccgagcaaga cctggagaag aaggtggagt actccttcct gctgggtccg 900

ctgctgggct tcgtggtgta cgagatcctg gagaacaccg ccgtgcagca catcaagaat 960

cagatcgacg agatcaagaa gcagctggac tccgcccagc acgacctgga ccgcgacgtg 1020

aagatcatcg ggatgctgaa cagcatcaac accgacatcg acaacctgta cagccaaggc 1080

caagaggcca tcaaggtgtt ccagaagctc caaggcatct gggcgaccat cggcgcgcag 1140

atcgagaacc tgaggaccac cagcctccaa gaggtccaag acagcgacga cgccgacgag 1200

atccagatcg agctggagga cgccagcgac gcgtggctgg tggtggccca agaggcgagg 1260

gacttcaccc tgaacgcgta cagcaccaac tcccgtcaga acctgccgat caacgtgatc 1320

agcgacagct gcaactgctc caccaccaac atgaccagca accagtactc caacccgacc 1380

accaacatga ccagcaacca gtacatgatc tcccacgagt acaccagcct gccgaacaac 1440

ttcatgctgt cccgcaactc taacctggag tacaagtgcc cggagaacaa cttcatgatc 1500

tactggtaca acaactccga ctggtacaac aactcagact ggtacaacaa ctga 1554

<210> 17

<211> 517

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 17

Met Ala His Ala Arg Val Leu Leu Leu Ala Leu Ala Val Leu Ala Thr

1 5 10 15

Ala Ala Val Ala Val Ala Ser Ser Ser Ser Phe Ala Asp Ser Asn Pro

20 25 30

Ile Arg Pro Val Thr Asp Arg Ala Ala Ser Thr Ile Ile Asp Ser Lys

35 40 45

Thr Thr Leu Pro Arg His Ser Leu Ile His Thr Ile Lys Leu Asn Ser

50 55 60

Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr Asn Gly Asn Gln Phe Ile

65 70 75 80

Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly

85 90 95

Leu Gly Leu Pro Val Asn Glu Gln Gln Leu Arg Thr His Val Asn Leu

100 105 110

Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val

115 120 125

Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr

130 135 140

Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys

145 150 155 160

Val Ala Gly Asp Pro Ser Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu

165 170 175

Gln Asp Glu Leu Asp Asn Ile Val Asp Asn Asn Ser Asp Asp Asp Ala

180 185 190

Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile

195 200 205

Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala Lys Asn Ile Val Thr Ser

210 215 220

Leu Asp Gln Phe Leu His Gly Asp Gln Lys Lys Leu Glu Gly Val Ile

225 230 235 240

Asn Ile Gln Lys Arg Leu Lys Glu Val Gln Thr Ala Leu Asn Gln Ala

245 250 255

His Gly Glu Ser Ser Pro Ala His Lys Glu Leu Leu Glu Lys Val Lys

260 265 270

Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu

275 280 285

Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe

290 295 300

Val Val Tyr Glu Ile Leu Glu Asn Thr Ala Val Gln His Ile Lys Asn

305 310 315 320

Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp Ser Ala Gln His Asp Leu

325 330 335

Asp Arg Asp Val Lys Ile Ile Gly Met Leu Asn Ser Ile Asn Thr Asp

340 345 350

Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu Ala Ile Lys Val Phe Gln

355 360 365

Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu

370 375 380

Arg Thr Thr Ser Leu Gln Glu Val Gln Asp Ser Asp Asp Ala Asp Glu

385 390 395 400

Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp Ala Trp Leu Val Val Ala

405 410 415

Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg

420 425 430

Gln Asn Leu Pro Ile Asn Val Ile Ser Asp Ser Cys Asn Cys Ser Thr

435 440 445

Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr

450 455 460

Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn

465 470 475 480

Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn

485 490 495

Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser

500 505 510

Asp Trp Tyr Asn Asn

515

<210> 18

<211> 1524

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 18

atgagtagct caccgtcctt tggcctgctg gcggttgcgg ccctgcttct ggcgcttagc 60

ctggcgcaac acggcatcat tgactccaag accaccctgc cgcgtcacag cctgatccac 120

accatcaagc tgaactccaa caagaagtac ggacctggcg acatgaccaa cggcaaccag 180

ttcatcatct ccaagcaaga gtgggccacc atcggagcgt acatccagac cggcctgggc 240

ctgccggtga acgagcagca gctgaggact cacgtgaacc tgtcccaaga catcagcatc 300

ccgtccgact tctcccagct gtacgacgtg tattgcagcg acaagacctc cgccgagtgg 360

tggaacaaga acctgtaccc gctgatcatc aagagcgcca acgacatcgc ctcctacggc 420

ttcaaggtgg cgggcgaccc gagcatcaag aaggacggct acttcaagaa gctccaagac 480

gagctggaca acatcgtgga caacaactcc gacgacgacg ccatcgccaa ggcgatcaag 540

gacttcaagg cgaggtgcgg catcctgatc aaggaggcga agcagtacga ggaggcggcg 600

aagaacatcg tgacctccct ggaccagttc ctgcacggcg accagaagaa gctggagggc 660

gtgatcaaca tccagaagag gctgaaggag gtgcagaccg ccctgaatca agcccacggc 720

gagagcagcc cggcgcacaa ggagctgctg gagaaggtga agaacctcaa gaccaccctg 780

gagaggacca tcaaggccga gcaagacctg gagaagaagg tggagtactc cttcctgctg 840

ggtccgctgc tgggcttcgt ggtgtacgag atcctggaga acaccgccgt gcagcacatc 900

aagaatcaga tcgacgagat caagaagcag ctggactccg cccagcacga cctggaccgc 960

gacgtgaaga tcatcgggat gctgaacagc atcaacaccg acatcgacaa cctgtacagc 1020

caaggccaag aggccatcaa ggtgttccag aagctccaag gcatctgggc gaccatcggc 1080

gcgcagatcg agaacctgag gaccaccagc ctccaagagg tccaagacag cgacgacgcc 1140

gacgagatcc agatcgagct ggaggacgcc agcgacgcgt ggctggtggt ggcccaagag 1200

gcgagggact tcaccctgaa cgcgtacagc accaactccc gtcagaacct gccgatcaac 1260

gtgatcagcg acagctgcaa ctgctccacc accaacatga ccagcaacca gtactccaac 1320

ccgaccacca acatgaccag caaccagtac atgatctccc acgagtacac cagcctgccg 1380

aacaacttca tgctgtcccg caactctaac ctggagtaca agtgcccgga gaacaacttc 1440

atgatctact ggtacaacaa ctccgactgg tacaacaact cagactggta caacaacgac 1500

gagctgaagg ctgaggccaa gtga 1524

<210> 19

<211> 507

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 19

Met Ser Ser Ser Pro Ser Phe Gly Leu Leu Ala Val Ala Ala Leu Leu

1 5 10 15

Leu Ala Leu Ser Leu Ala Gln His Gly Ile Ile Asp Ser Lys Thr Thr

20 25 30

Leu Pro Arg His Ser Leu Ile His Thr Ile Lys Leu Asn Ser Asn Lys

35 40 45

Lys Tyr Gly Pro Gly Asp Met Thr Asn Gly Asn Gln Phe Ile Ile Ser

50 55 60

Lys Gln Glu Trp Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly

65 70 75 80

Leu Pro Val Asn Glu Gln Gln Leu Arg Thr His Val Asn Leu Ser Gln

85 90 95

Asp Ile Ser Ile Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys

100 105 110

Ser Asp Lys Thr Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu

115 120 125

Ile Ile Lys Ser Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala

130 135 140

Gly Asp Pro Ser Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp

145 150 155 160

Glu Leu Asp Asn Ile Val Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala

165 170 175

Lys Ala Ile Lys Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu

180 185 190

Ala Lys Gln Tyr Glu Glu Ala Ala Lys Asn Ile Val Thr Ser Leu Asp

195 200 205

Gln Phe Leu His Gly Asp Gln Lys Lys Leu Glu Gly Val Ile Asn Ile

210 215 220

Gln Lys Arg Leu Lys Glu Val Gln Thr Ala Leu Asn Gln Ala His Gly

225 230 235 240

Glu Ser Ser Pro Ala His Lys Glu Leu Leu Glu Lys Val Lys Asn Leu

245 250 255

Lys Thr Thr Leu Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys

260 265 270

Lys Val Glu Tyr Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe Val Val

275 280 285

Tyr Glu Ile Leu Glu Asn Thr Ala Val Gln His Ile Lys Asn Gln Ile

290 295 300

Asp Glu Ile Lys Lys Gln Leu Asp Ser Ala Gln His Asp Leu Asp Arg

305 310 315 320

Asp Val Lys Ile Ile Gly Met Leu Asn Ser Ile Asn Thr Asp Ile Asp

325 330 335

Asn Leu Tyr Ser Gln Gly Gln Glu Ala Ile Lys Val Phe Gln Lys Leu

340 345 350

Gln Gly Ile Trp Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr

355 360 365

Thr Ser Leu Gln Glu Val Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln

370 375 380

Ile Glu Leu Glu Asp Ala Ser Asp Ala Trp Leu Val Val Ala Gln Glu

385 390 395 400

Ala Arg Asp Phe Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg Gln Asn

405 410 415

Leu Pro Ile Asn Val Ile Ser Asp Ser Cys Asn Cys Ser Thr Thr Asn

420 425 430

Met Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn

435 440 445

Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met

450 455 460

Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe

465 470 475 480

Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp

485 490 495

Tyr Asn Asn Asp Glu Leu Lys Ala Glu Ala Lys

500 505

<210> 20

<211> 1707

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 20

atggcgaaca agcacctgag cctgtccctg ttcctggtgc tgcttggcct gagcgcatcc 60

ctggcatccg gaatcattga ctccaagacc accctgccgc gtcacagcct gatccacacc 120

atcaagctga actccaacaa gaagtacgga cctggcgaca tgaccaacgg caaccagttc 180

atcatctcca agcaagagtg ggccaccatc ggagcgtaca tccagaccgg cctgggcctg 240

ccggtgaacg agcagcagct gaggactcac gtgaacctgt cccaagacat cagcatcccg 300

tccgacttct cccagctgta cgacgtgtat tgcagcgaca agacctccgc cgagtggtgg 360

aacaagaacc tgtacccgct gatcatcaag agcgccaacg acatcgcctc ctacggcttc 420

aaggtggcgg gcgacccgag catcaagaag gacggctact tcaagaagct ccaagacgag 480

ctggacaaca tcgtggacaa caactccgac gacgacgcca tcgccaaggc gatcaaggac 540

ttcaaggcga ggtgcggcat cctgatcaag gaggcgaagc agtacgagga ggcggcgaag 600

aacatcgtga cctccctgga ccagttcctg cacggcgacc agaagaagct ggagggcgtg 660

atcaacatcc agaagaggct gaaggaggtg cagaccgccc tgaatcaagc ccacggcgag 720

agcagcccgg cgcacaagga gctgctggag aaggtgaaga acctcaagac caccctggag 780

aggaccatca aggccgagca agacctggag aagaaggtgg agtactcctt cctgctgggt 840

ccgctgctgg gcttcgtggt gtacgagatc ctggagaaca ccgccgtgca gcacatcaag 900

aatcagatcg acgagatcaa gaagcagctg gactccgccc agcacgacct ggaccgcgac 960

gtgaagatca tcgggatgct gaacagcatc aacaccgaca tcgacaacct gtacagccaa 1020

ggccaagagg ccatcaaggt gttccagaag ctccaaggca tctgggcgac catcggcgcg 1080

cagatcgaga acctgaggac caccagcctc caagaggtcc aagacagcga cgacgccgac 1140

gagatccaga tcgagctgga ggacgccagc gacgcgtggc tggtggtggc ccaagaggcg 1200

agggacttca ccctgaacgc gtacagcacc aactcccgtc agaacctgcc gatcaacgtg 1260

atcagcgaca gctgcaactg ctccaccacc aacatgacca gcaaccagta ctccaacccg 1320

accaccaaca tgaccagcaa ccagtacatg atctcccacg agtacaccag cctgccgaac 1380

aacttcatgc tgtcccgcaa ctctaacctg gagtacaagt gcccggagaa caacttcatg 1440

atctactggt acaacaactc cgactggtac aacaactcag actggtacaa caactctaag 1500

accgcagtgc aagccaaggc agcgtgggct gccgtgtggg gaatcctgat cgtggtggca 1560

gtggtggctg ctggcagcta cgtggtgtac aagtataggc tgaggagcta catggactca 1620

gagatcagag cgatcatggc acagtacatg cctctggaca accaaggtga ggtgccgaac 1680

cacacccacg acgaggaccg gagctga 1707

<210> 21

<211> 568

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 21

Met Ala Asn Lys His Leu Ser Leu Ser Leu Phe Leu Val Leu Leu Gly

1 5 10 15

Leu Ser Ala Ser Leu Ala Ser Gly Ile Ile Asp Ser Lys Thr Thr Leu

20 25 30

Pro Arg His Ser Leu Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys

35 40 45

Tyr Gly Pro Gly Asp Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys

50 55 60

Gln Glu Trp Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu

65 70 75 80

Pro Val Asn Glu Gln Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp

85 90 95

Ile Ser Ile Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser

100 105 110

Asp Lys Thr Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile

115 120 125

Ile Lys Ser Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly

130 135 140

Asp Pro Ser Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu

145 150 155 160

Leu Asp Asn Ile Val Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys

165 170 175

Ala Ile Lys Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala

180 185 190

Lys Gln Tyr Glu Glu Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln

195 200 205

Phe Leu His Gly Asp Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln

210 215 220

Lys Arg Leu Lys Glu Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu

225 230 235 240

Ser Ser Pro Ala His Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys

245 250 255

Thr Thr Leu Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys

260 265 270

Val Glu Tyr Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr

275 280 285

Glu Ile Leu Glu Asn Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp

290 295 300

Glu Ile Lys Lys Gln Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp

305 310 315 320

Val Lys Ile Ile Gly Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn

325 330 335

Leu Tyr Ser Gln Gly Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln

340 345 350

Gly Ile Trp Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr

355 360 365

Ser Leu Gln Glu Val Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile

370 375 380

Glu Leu Glu Asp Ala Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala

385 390 395 400

Arg Asp Phe Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg Gln Asn Leu

405 410 415

Pro Ile Asn Val Ile Ser Asp Ser Cys Asn Cys Ser Thr Thr Asn Met

420 425 430

Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln

435 440 445

Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu

450 455 460

Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met

465 470 475 480

Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr

485 490 495

Asn Asn Ser Lys Thr Ala Val Gln Ala Lys Ala Ala Trp Ala Ala Val

500 505 510

Trp Gly Ile Leu Ile Val Val Ala Val Val Ala Ala Gly Ser Tyr Val

515 520 525

Val Tyr Lys Tyr Arg Leu Arg Ser Tyr Met Asp Ser Glu Ile Arg Ala

530 535 540

Ile Met Ala Gln Tyr Met Pro Leu Asp Asn Gln Gly Glu Val Pro Asn

545 550 555 560

His Thr His Asp Glu Asp Arg Ser

565

<210> 22

<211> 1497

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 22

atggcgaaca agcacctgag cctgtccctg ttcctggtgc tgcttggcct gagcgcatcc 60

ctggcatccg gaatcattga ctccaagacc accctgccgc gtcacagcct gatccacacc 120

atcaagctga actccaacaa gaagtacgga cctggcgaca tgaccaacgg caaccagttc 180

atcatctcca agcaagagtg ggccaccatc ggagcgtaca tccagaccgg cctgggcctg 240

ccggtgaacg agcagcagct gaggactcac gtgaacctgt cccaagacat cagcatcccg 300

tccgacttct cccagctgta cgacgtgtat tgcagcgaca agacctccgc cgagtggtgg 360

aacaagaacc tgtacccgct gatcatcaag agcgccaacg acatcgcctc ctacggcttc 420

aaggtggcgg gcgacccgag catcaagaag gacggctact tcaagaagct ccaagacgag 480

ctggacaaca tcgtggacaa caactccgac gacgacgcca tcgccaaggc gatcaaggac 540

ttcaaggcga ggtgcggcat cctgatcaag gaggcgaagc agtacgagga ggcggcgaag 600

aacatcgtga cctccctgga ccagttcctg cacggcgacc agaagaagct ggagggcgtg 660

atcaacatcc agaagaggct gaaggaggtg cagaccgccc tgaatcaagc ccacggcgag 720

agcagcccgg cgcacaagga gctgctggag aaggtgaaga acctcaagac caccctggag 780

aggaccatca aggccgagca agacctggag aagaaggtgg agtactcctt cctgctgggt 840

ccgctgctgg gcttcgtggt gtacgagatc ctggagaaca ccgccgtgca gcacatcaag 900

aatcagatcg acgagatcaa gaagcagctg gactccgccc agcacgacct ggaccgcgac 960

gtgaagatca tcgggatgct gaacagcatc aacaccgaca tcgacaacct gtacagccaa 1020

ggccaagagg ccatcaaggt gttccagaag ctccaaggca tctgggcgac catcggcgcg 1080

cagatcgaga acctgaggac caccagcctc caagaggtcc aagacagcga cgacgccgac 1140

gagatccaga tcgagctgga ggacgccagc gacgcgtggc tggtggtggc ccaagaggcg 1200

agggacttca ccctgaacgc gtacagcacc aactcccgtc agaacctgcc gatcaacgtg 1260

atcagcgaca gctgcaactg ctccaccacc aacatgacca gcaaccagta ctccaacccg 1320

accaccaaca tgaccagcaa ccagtacatg atctcccacg agtacaccag cctgccgaac 1380

aacttcatgc tgtcccgcaa ctctaacctg gagtacaagt gcccggagaa caacttcatg 1440

atctactggt acaacaactc cgactggtac aacaactcag actggtacaa caactga 1497

<210> 23

<211> 498

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 23

Met Ala Asn Lys His Leu Ser Leu Ser Leu Phe Leu Val Leu Leu Gly

1 5 10 15

Leu Ser Ala Ser Leu Ala Ser Gly Ile Ile Asp Ser Lys Thr Thr Leu

20 25 30

Pro Arg His Ser Leu Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys

35 40 45

Tyr Gly Pro Gly Asp Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys

50 55 60

Gln Glu Trp Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu

65 70 75 80

Pro Val Asn Glu Gln Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp

85 90 95

Ile Ser Ile Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser

100 105 110

Asp Lys Thr Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile

115 120 125

Ile Lys Ser Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly

130 135 140

Asp Pro Ser Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu

145 150 155 160

Leu Asp Asn Ile Val Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys

165 170 175

Ala Ile Lys Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala

180 185 190

Lys Gln Tyr Glu Glu Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln

195 200 205

Phe Leu His Gly Asp Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln

210 215 220

Lys Arg Leu Lys Glu Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu

225 230 235 240

Ser Ser Pro Ala His Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys

245 250 255

Thr Thr Leu Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys

260 265 270

Val Glu Tyr Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr

275 280 285

Glu Ile Leu Glu Asn Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp

290 295 300

Glu Ile Lys Lys Gln Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp

305 310 315 320

Val Lys Ile Ile Gly Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn

325 330 335

Leu Tyr Ser Gln Gly Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln

340 345 350

Gly Ile Trp Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr

355 360 365

Ser Leu Gln Glu Val Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile

370 375 380

Glu Leu Glu Asp Ala Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala

385 390 395 400

Arg Asp Phe Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg Gln Asn Leu

405 410 415

Pro Ile Asn Val Ile Ser Asp Ser Cys Asn Cys Ser Thr Thr Asn Met

420 425 430

Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln

435 440 445

Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu

450 455 460

Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met

465 470 475 480

Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr

485 490 495

Asn Asn

<210> 24

<211> 1515

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 24

atggcgaaca agcacctgag cctgtccctg ttcctggtgc tgcttggcct gagcgcatcc 60

ctggcatccg gaatcattga ctccaagacc accctgccgc gtcacagcct gatccacacc 120

atcaagctga actccaacaa gaagtacgga cctggcgaca tgaccaacgg caaccagttc 180

atcatctcca agcaagagtg ggccaccatc ggagcgtaca tccagaccgg cctgggcctg 240

ccggtgaacg agcagcagct gaggactcac gtgaacctgt cccaagacat cagcatcccg 300

tccgacttct cccagctgta cgacgtgtat tgcagcgaca agacctccgc cgagtggtgg 360

aacaagaacc tgtacccgct gatcatcaag agcgccaacg acatcgcctc ctacggcttc 420

aaggtggcgg gcgacccgag catcaagaag gacggctact tcaagaagct ccaagacgag 480

ctggacaaca tcgtggacaa caactccgac gacgacgcca tcgccaaggc gatcaaggac 540

ttcaaggcga ggtgcggcat cctgatcaag gaggcgaagc agtacgagga ggcggcgaag 600

aacatcgtga cctccctgga ccagttcctg cacggcgacc agaagaagct ggagggcgtg 660

atcaacatcc agaagaggct gaaggaggtg cagaccgccc tgaatcaagc ccacggcgag 720

agcagcccgg cgcacaagga gctgctggag aaggtgaaga acctcaagac caccctggag 780

aggaccatca aggccgagca agacctggag aagaaggtgg agtactcctt cctgctgggt 840

ccgctgctgg gcttcgtggt gtacgagatc ctggagaaca ccgccgtgca gcacatcaag 900

aatcagatcg acgagatcaa gaagcagctg gactccgccc agcacgacct ggaccgcgac 960

gtgaagatca tcgggatgct gaacagcatc aacaccgaca tcgacaacct gtacagccaa 1020

ggccaagagg ccatcaaggt gttccagaag ctccaaggca tctgggcgac catcggcgcg 1080

cagatcgaga acctgaggac caccagcctc caagaggtcc aagacagcga cgacgccgac 1140

gagatccaga tcgagctgga ggacgccagc gacgcgtggc tggtggtggc ccaagaggcg 1200

agggacttca ccctgaacgc gtacagcacc aactcccgtc agaacctgcc gatcaacgtg 1260

atcagcgaca gctgcaactg ctccaccacc aacatgacca gcaaccagta ctccaacccg 1320

accaccaaca tgaccagcaa ccagtacatg atctcccacg agtacaccag cctgccgaac 1380

aacttcatgc tgtcccgcaa ctctaacctg gagtacaagt gcccggagaa caacttcatg 1440

atctactggt acaacaactc cgactggtac aacaactcag actggtacaa caacagcgag 1500

aaggacgagc tgtga 1515

<210> 25

<211> 504

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 25

Met Ala Asn Lys His Leu Ser Leu Ser Leu Phe Leu Val Leu Leu Gly

1 5 10 15

Leu Ser Ala Ser Leu Ala Ser Gly Ile Ile Asp Ser Lys Thr Thr Leu

20 25 30

Pro Arg His Ser Leu Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys

35 40 45

Tyr Gly Pro Gly Asp Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys

50 55 60

Gln Glu Trp Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu

65 70 75 80

Pro Val Asn Glu Gln Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp

85 90 95

Ile Ser Ile Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser

100 105 110

Asp Lys Thr Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile

115 120 125

Ile Lys Ser Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly

130 135 140

Asp Pro Ser Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu

145 150 155 160

Leu Asp Asn Ile Val Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys

165 170 175

Ala Ile Lys Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala

180 185 190

Lys Gln Tyr Glu Glu Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln

195 200 205

Phe Leu His Gly Asp Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln

210 215 220

Lys Arg Leu Lys Glu Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu

225 230 235 240

Ser Ser Pro Ala His Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys

245 250 255

Thr Thr Leu Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys

260 265 270

Val Glu Tyr Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr

275 280 285

Glu Ile Leu Glu Asn Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp

290 295 300

Glu Ile Lys Lys Gln Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp

305 310 315 320

Val Lys Ile Ile Gly Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn

325 330 335

Leu Tyr Ser Gln Gly Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln

340 345 350

Gly Ile Trp Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr

355 360 365

Ser Leu Gln Glu Val Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile

370 375 380

Glu Leu Glu Asp Ala Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala

385 390 395 400

Arg Asp Phe Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg Gln Asn Leu

405 410 415

Pro Ile Asn Val Ile Ser Asp Ser Cys Asn Cys Ser Thr Thr Asn Met

420 425 430

Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln

435 440 445

Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu

450 455 460

Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met

465 470 475 480

Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr

485 490 495

Asn Asn Ser Glu Lys Asp Glu Leu

500

<210> 26

<211> 1488

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 26

atggcaccga gaaagaggaa ggagagcaat agggagtcag cgaggaggag cagatacaga 60

aagatcattg actccaagac caccctgccg cgtcacagcc tgatccacac catcaagctg 120

aactccaaca agaagtacgg acctggcgac atgaccaacg gcaaccagtt catcatctcc 180

aagcaagagt gggccaccat cggagcgtac atccagaccg gcctgggcct gccggtgaac 240

gagcagcagc tgaggactca cgtgaacctg tcccaagaca tcagcatccc gtccgacttc 300

tcccagctgt acgacgtgta ttgcagcgac aagacctccg ccgagtggtg gaacaagaac 360

ctgtacccgc tgatcatcaa gagcgccaac gacatcgcct cctacggctt caaggtggcg 420

ggcgacccga gcatcaagaa ggacggctac ttcaagaagc tccaagacga gctggacaac 480

atcgtggaca acaactccga cgacgacgcc atcgccaagg cgatcaagga cttcaaggcg 540

aggtgcggca tcctgatcaa ggaggcgaag cagtacgagg aggcggcgaa gaacatcgtg 600

acctccctgg accagttcct gcacggcgac cagaagaagc tggagggcgt gatcaacatc 660

cagaagaggc tgaaggaggt gcagaccgcc ctgaatcaag cccacggcga gagcagcccg 720

gcgcacaagg agctgctgga gaaggtgaag aacctcaaga ccaccctgga gaggaccatc 780

aaggccgagc aagacctgga gaagaaggtg gagtactcct tcctgctggg tccgctgctg 840

ggcttcgtgg tgtacgagat cctggagaac accgccgtgc agcacatcaa gaatcagatc 900

gacgagatca agaagcagct ggactccgcc cagcacgacc tggaccgcga cgtgaagatc 960

atcgggatgc tgaacagcat caacaccgac atcgacaacc tgtacagcca aggccaagag 1020

gccatcaagg tgttccagaa gctccaaggc atctgggcga ccatcggcgc gcagatcgag 1080

aacctgagga ccaccagcct ccaagaggtc caagacagcg acgacgccga cgagatccag 1140

atcgagctgg aggacgccag cgacgcgtgg ctggtggtgg cccaagaggc gagggacttc 1200

accctgaacg cgtacagcac caactcccgt cagaacctgc cgatcaacgt gatcagcgac 1260

agctgcaact gctccaccac caacatgacc agcaaccagt actccaaccc gaccaccaac 1320

atgaccagca accagtacat gatctcccac gagtacacca gcctgccgaa caacttcatg 1380

ctgtcccgca actctaacct ggagtacaag tgcccggaga acaacttcat gatctactgg 1440

tacaacaact ccgactggta caacaactca gactggtaca acaactga 1488

<210> 27

<211> 495

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 27

Met Ala Pro Arg Lys Arg Lys Glu Ser Asn Arg Glu Ser Ala Arg Arg

1 5 10 15

Ser Arg Tyr Arg Lys Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His

20 25 30

Ser Leu Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro

35 40 45

Gly Asp Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp

50 55 60

Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn

65 70 75 80

Glu Gln Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile

85 90 95

Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr

100 105 110

Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser

115 120 125

Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser

130 135 140

Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn

145 150 155 160

Ile Val Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys

165 170 175

Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr

180 185 190

Glu Glu Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His

195 200 205

Gly Asp Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu

210 215 220

Lys Glu Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro

225 230 235 240

Ala His Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu

245 250 255

Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr

260 265 270

Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu

275 280 285

Glu Asn Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys

290 295 300

Lys Gln Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile

305 310 315 320

Ile Gly Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser

325 330 335

Gln Gly Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp

340 345 350

Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln

355 360 365

Glu Val Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu

370 375 380

Asp Ala Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe

385 390 395 400

Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn

405 410 415

Val Ile Ser Asp Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn

420 425 430

Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile

435 440 445

Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn

450 455 460

Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp

465 470 475 480

Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

485 490 495

<210> 28

<211> 1482

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 28

atgatcattg actccaagac caccctgccg cgtcacagcc tgatccacac catcaagctg 60

aactccaaca agaagtacgg acctggcgac atgaccaacg gcaaccagtt catcatctcc 120

aagcaagagt gggccaccat cggagcgtac atccagaccg gcctgggcct gccggtgaac 180

gagcagcagc tgaggactca cgtgaacctg tcccaagaca tcagcatccc gtccgacttc 240

tcccagctgt acgacgtgta ttgcagcgac aagacctccg ccgagtggtg gaacaagaac 300

ctgtacccgc tgatcatcaa gagcgccaac gacatcgcct cctacggctt caaggtggcg 360

ggcgacccga gcatcaagaa ggacggctac ttcaagaagc tccaagacga gctggacaac 420

atcgtggaca acaactccga cgacgacgcc atcgccaagg cgatcaagga cttcaaggcg 480

aggtgcggca tcctgatcaa ggaggcgaag cagtacgagg aggcggcgaa gaacatcgtg 540

acctccctgg accagttcct gcacggcgac cagaagaagc tggagggcgt gatcaacatc 600

cagaagaggc tgaaggaggt gcagaccgcc ctgaatcaag cccacggcga gagcagcccg 660

gcgcacaagg agctgctgga gaaggtgaag aacctcaaga ccaccctgga gaggaccatc 720

aaggccgagc aagacctgga gaagaaggtg gagtactcct tcctgctggg tccgctgctg 780

ggcttcgtgg tgtacgagat cctggagaac accgccgtgc agcacatcaa gaatcagatc 840

gacgagatca agaagcagct ggactccgcc cagcacgacc tggaccgcga cgtgaagatc 900

atcgggatgc tgaacagcat caacaccgac atcgacaacc tgtacagcca aggccaagag 960

gccatcaagg tgttccagaa gctccaaggc atctgggcga ccatcggcgc gcagatcgag 1020

aacctgagga ccaccagcct ccaagaggtc caagacagcg acgacgccga cgagatccag 1080

atcgagctgg aggacgccag cgacgcgtgg ctggtggtgg cccaagaggc gagggacttc 1140

accctgaacg cgtacagcac caactcccgt cagaacctgc cgatcaacgt gatcagcgac 1200

agctgcaact gctccaccac caacatgacc agcaaccagt actccaaccc gaccaccaac 1260

atgaccagca accagtacat gatctcccac gagtacacca gcctgccgaa caacttcatg 1320

ctgtcccgca actctaacct ggagtacaag tgcccggaga acaacttcat gatctactgg 1380

tacaacaact ccgactggta caacaactca gactggtaca acaacagaaa gaggaaggag 1440

agcaataggg agtcagcgag gaggagcaga tacagaaagt ga 1482

<210> 29

<211> 493

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 29

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Arg Lys Arg Lys Glu

465 470 475 480

Ser Asn Arg Glu Ser Ala Arg Arg Ser Arg Tyr Arg Lys

485 490

<210> 30

<211> 1641

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 30

atgttggctc gccaaggagg atcactgaga gcctctcagt gtaacgctgg cctcgcgaga 60

cgcgtggagg tgggagcgtt ggttgttccg agacccataa gcgtcaacga cgtggttccc 120

catgtctatt cggctcctct gagcgtcgcg aggaggtcgt gctccaagtc atccatccgc 180

tcgactcgca gacttcagac aaccgtctgc tccatgatca ttgactccaa gaccaccctg 240

ccgcgtcaca gcctgatcca caccatcaag ctgaactcca acaagaagta cggacctggc 300

gacatgacca acggcaacca gttcatcatc tccaagcaag agtgggccac catcggagcg 360

tacatccaga ccggcctggg cctgccggtg aacgagcagc agctgaggac tcacgtgaac 420

ctgtcccaag acatcagcat cccgtccgac ttctcccagc tgtacgacgt gtattgcagc 480

gacaagacct ccgccgagtg gtggaacaag aacctgtacc cgctgatcat caagagcgcc 540

aacgacatcg cctcctacgg cttcaaggtg gcgggcgacc cgagcatcaa gaaggacggc 600

tacttcaaga agctccaaga cgagctggac aacatcgtgg acaacaactc cgacgacgac 660

gccatcgcca aggcgatcaa ggacttcaag gcgaggtgcg gcatcctgat caaggaggcg 720

aagcagtacg aggaggcggc gaagaacatc gtgacctccc tggaccagtt cctgcacggc 780

gaccagaaga agctggaggg cgtgatcaac atccagaaga ggctgaagga ggtgcagacc 840

gccctgaatc aagcccacgg cgagagcagc ccggcgcaca aggagctgct ggagaaggtg 900

aagaacctca agaccaccct ggagaggacc atcaaggccg agcaagacct ggagaagaag 960

gtggagtact ccttcctgct gggtccgctg ctgggcttcg tggtgtacga gatcctggag 1020

aacaccgccg tgcagcacat caagaatcag atcgacgaga tcaagaagca gctggactcc 1080

gcccagcacg acctggaccg cgacgtgaag atcatcggga tgctgaacag catcaacacc 1140

gacatcgaca acctgtacag ccaaggccaa gaggccatca aggtgttcca gaagctccaa 1200

ggcatctggg cgaccatcgg cgcgcagatc gagaacctga ggaccaccag cctccaagag 1260

gtccaagaca gcgacgacgc cgacgagatc cagatcgagc tggaggacgc cagcgacgcg 1320

tggctggtgg tggcccaaga ggcgagggac ttcaccctga acgcgtacag caccaactcc 1380

cgtcagaacc tgccgatcaa cgtgatcagc gacagctgca actgctccac caccaacatg 1440

accagcaacc agtactccaa cccgaccacc aacatgacca gcaaccagta catgatctcc 1500

cacgagtaca ccagcctgcc gaacaacttc atgctgtccc gcaactctaa cctggagtac 1560

aagtgcccgg agaacaactt catgatctac tggtacaaca actccgactg gtacaacaac 1620

tcagactggt acaacaactg a 1641

<210> 31

<211> 546

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 31

Met Leu Ala Arg Gln Gly Gly Ser Leu Arg Ala Ser Gln Cys Asn Ala

1 5 10 15

Gly Leu Ala Arg Arg Val Glu Val Gly Ala Leu Val Val Pro Arg Pro

20 25 30

Ile Ser Val Asn Asp Val Val Pro His Val Tyr Ser Ala Pro Leu Ser

35 40 45

Val Ala Arg Arg Ser Cys Ser Lys Ser Ser Ile Arg Ser Thr Arg Arg

50 55 60

Leu Gln Thr Thr Val Cys Ser Met Ile Ile Asp Ser Lys Thr Thr Leu

65 70 75 80

Pro Arg His Ser Leu Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys

85 90 95

Tyr Gly Pro Gly Asp Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys

100 105 110

Gln Glu Trp Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu

115 120 125

Pro Val Asn Glu Gln Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp

130 135 140

Ile Ser Ile Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser

145 150 155 160

Asp Lys Thr Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile

165 170 175

Ile Lys Ser Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly

180 185 190

Asp Pro Ser Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu

195 200 205

Leu Asp Asn Ile Val Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys

210 215 220

Ala Ile Lys Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala

225 230 235 240

Lys Gln Tyr Glu Glu Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln

245 250 255

Phe Leu His Gly Asp Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln

260 265 270

Lys Arg Leu Lys Glu Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu

275 280 285

Ser Ser Pro Ala His Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys

290 295 300

Thr Thr Leu Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys

305 310 315 320

Val Glu Tyr Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr

325 330 335

Glu Ile Leu Glu Asn Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp

340 345 350

Glu Ile Lys Lys Gln Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp

355 360 365

Val Lys Ile Ile Gly Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn

370 375 380

Leu Tyr Ser Gln Gly Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln

385 390 395 400

Gly Ile Trp Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr

405 410 415

Ser Leu Gln Glu Val Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile

420 425 430

Glu Leu Glu Asp Ala Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala

435 440 445

Arg Asp Phe Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg Gln Asn Leu

450 455 460

Pro Ile Asn Val Ile Ser Asp Ser Cys Asn Cys Ser Thr Thr Asn Met

465 470 475 480

Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln

485 490 495

Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu

500 505 510

Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met

515 520 525

Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr

530 535 540

Asn Asn

545

<210> 32

<211> 1626

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 32

atggctcaat ctagcagaat ctgccacggt gtgcagaacc catgtgtgat catttccaat 60

ctctccaaat ccaaccagaa caaatctcct ttctcagtca gcctcaagac tcaccagcag 120

cagcgtcgtg cttaccagat atctagctgg ggattgaaga agtcaaacaa cgggtccgtg 180

attcgtccgg ttaaggcaat gatcattgac tccaagacca ccctgccgcg tcacagcctg 240

atccacacca tcaagctgaa ctccaacaag aagtacggac ctggcgacat gaccaacggc 300

aaccagttca tcatctccaa gcaagagtgg gccaccatcg gagcgtacat ccagaccggc 360

ctgggcctgc cggtgaacga gcagcagctg aggactcacg tgaacctgtc ccaagacatc 420

agcatcccgt ccgacttctc ccagctgtac gacgtgtatt gcagcgacaa gacctccgcc 480

gagtggtgga acaagaacct gtacccgctg atcatcaaga gcgccaacga catcgcctcc 540

tacggcttca aggtggcggg cgacccgagc atcaagaagg acggctactt caagaagctc 600

caagacgagc tggacaacat cgtggacaac aactccgacg acgacgccat cgccaaggcg 660

atcaaggact tcaaggcgag gtgcggcatc ctgatcaagg aggcgaagca gtacgaggag 720

gcggcgaaga acatcgtgac ctccctggac cagttcctgc acggcgacca gaagaagctg 780

gagggcgtga tcaacatcca gaagaggctg aaggaggtgc agaccgccct gaatcaagcc 840

cacggcgaga gcagcccggc gcacaaggag ctgctggaga aggtgaagaa cctcaagacc 900

accctggaga ggaccatcaa ggccgagcaa gacctggaga agaaggtgga gtactccttc 960

ctgctgggtc cgctgctggg cttcgtggtg tacgagatcc tggagaacac cgccgtgcag 1020

cacatcaaga atcagatcga cgagatcaag aagcagctgg actccgccca gcacgacctg 1080

gaccgcgacg tgaagatcat cgggatgctg aacagcatca acaccgacat cgacaacctg 1140

tacagccaag gccaagaggc catcaaggtg ttccagaagc tccaaggcat ctgggcgacc 1200

atcggcgcgc agatcgagaa cctgaggacc accagcctcc aagaggtcca agacagcgac 1260

gacgccgacg agatccagat cgagctggag gacgccagcg acgcgtggct ggtggtggcc 1320

caagaggcga gggacttcac cctgaacgcg tacagcacca actcccgtca gaacctgccg 1380

atcaacgtga tcagcgacag ctgcaactgc tccaccacca acatgaccag caaccagtac 1440

tccaacccga ccaccaacat gaccagcaac cagtacatga tctcccacga gtacaccagc 1500

ctgccgaaca acttcatgct gtcccgcaac tctaacctgg agtacaagtg cccggagaac 1560

aacttcatga tctactggta caacaactcc gactggtaca acaactcaga ctggtacaac 1620

aactga 1626

<210> 33

<211> 541

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 33

Met Ala Gln Ser Ser Arg Ile Cys His Gly Val Gln Asn Pro Cys Val

1 5 10 15

Ile Ile Ser Asn Leu Ser Lys Ser Asn Gln Asn Lys Ser Pro Phe Ser

20 25 30

Val Ser Leu Lys Thr His Gln Gln Gln Arg Arg Ala Tyr Gln Ile Ser

35 40 45

Ser Trp Gly Leu Lys Lys Ser Asn Asn Gly Ser Val Ile Arg Pro Val

50 55 60

Lys Ala Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu

65 70 75 80

Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp

85 90 95

Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr

100 105 110

Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln

115 120 125

Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser

130 135 140

Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala

145 150 155 160

Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn

165 170 175

Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys

180 185 190

Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val

195 200 205

Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe

210 215 220

Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu

225 230 235 240

Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp

245 250 255

Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu

260 265 270

Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His

275 280 285

Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg

290 295 300

Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe

305 310 315 320

Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn

325 330 335

Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln

340 345 350

Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly

355 360 365

Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly

370 375 380

Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr

385 390 395 400

Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val

405 410 415

Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala

420 425 430

Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu

435 440 445

Asn Ala Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile

450 455 460

Ser Asp Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr

465 470 475 480

Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His

485 490 495

Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn

500 505 510

Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn

515 520 525

Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

530 535 540

<210> 34

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 34

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta tgcttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa tgtcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattga 1428

<210> 35

<400> 35

000

<210> 36

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 36

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttcttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa tgtcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattga 1428

<210> 37

<211> 475

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 37

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Ser Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

465 470 475

<210> 38

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 38

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgagctggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa tgtcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattga 1428

<210> 39

<211> 475

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 39

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Ala Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

465 470 475

<210> 40

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 40

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatctggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa tgtcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattga 1428

<210> 41

<211> 475

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 41

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Ser Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

465 470 475

<210> 42

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 42

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa gctcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattga 1428

<210> 43

<211> 475

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 43

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Ala Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

465 470 475

<210> 44

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 44

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa tctcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattga 1428

<210> 45

<211> 475

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 45

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Ser Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

465 470 475

<210> 46

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 46

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta tagttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa agtcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattga 1428

<210> 47

<211> 475

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 47

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Ser Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Ser Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

465 470 475

<210> 48

<211> 1428

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 48

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta tgcttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagaa atagtaattt agaatataaa gctcctgaaa ataattttat gatatattgg 1380

tataataatt cggattggta taataattcg gattggtata ataattga 1428

<210> 49

<211> 475

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 49

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Ala Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu

435 440 445

Tyr Lys Ala Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser

450 455 460

Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

465 470 475

<210> 50

<211> 379

<212> PRT

<213> 人工序列

<220>

<223> 蛋白酶产生的片段

<400> 50

His Ser Leu Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly

1 5 10 15

Pro Gly Asp Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu

20 25 30

Trp Ala Thr Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val

35 40 45

Asn Glu Gln Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser

50 55 60

Ile Pro Ser Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys

65 70 75 80

Thr Ser Ala Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys

85 90 95

Ser Ala Asn Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro

100 105 110

Ser Ile Lys Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp

115 120 125

Asn Ile Val Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile

130 135 140

Lys Asp Phe Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln

145 150 155 160

Tyr Glu Glu Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu

165 170 175

His Gly Asp Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg

180 185 190

Leu Lys Glu Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser

195 200 205

Pro Ala His Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr

210 215 220

Leu Glu Arg Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu

225 230 235 240

Tyr Ser Phe Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile

245 250 255

Leu Glu Asn Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile

260 265 270

Lys Lys Gln Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys

275 280 285

Ile Ile Gly Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr

290 295 300

Ser Gln Gly Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile

305 310 315 320

Trp Ala Thr Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu

325 330 335

Gln Glu Val Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu

340 345 350

Glu Asp Ala Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp

355 360 365

Phe Thr Leu Asn Ala Tyr Ser Thr Asn Ser Arg

370 375

<210> 51

<211> 32

<212> PRT

<213> 人工序列

<220>

<223> 蛋白酶产生的片段

<400> 51

Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr

1 5 10 15

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

20 25 30

<210> 52

<211> 25

<212> PRT

<213> 人工序列

<220>

<223> 蛋白酶产生的片段

<400> 52

Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp

1 5 10 15

Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

20 25

<210> 53

<211> 1332

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 53

atgattattg atagtaaaac gactttacct agacattcac ttattcatac aattaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctacgat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

ttatcaagat ag 1332

<210> 54

<211> 443

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 54

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg

435 440

<210> 55

<211> 1296

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 55

atgattattg atagtaaaac gactttacct agacattcac ttattcatac aattaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctacgat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatag 1296

<210> 56

<211> 431

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 56

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu

420 425 430

<210> 57

<211> 1236

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 57

atgattattg atagtaaaac gactttacct agacattcac ttattcatac aattaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctacgat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcatag 1236

<210> 58

<211> 411

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 58

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser

405 410

<210> 59

<211> 1203

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 59

atgattattg atagtaaaac gactttacct agacattcac ttattcatac aattaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctacgat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tag 1203

<210> 60

<211> 400

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 60

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

<210> 61

<211> 1173

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 61

atgattattg atagtaaaac gactttacct agacattcac ttattcatac aattaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctacgat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga tag 1173

<210> 62

<211> 390

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 62

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg

385 390

<210> 63

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 63

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga aatgttatat cagattcatg taattgttca 1200

acaacaaata tgacatcaaa tcaatacagt aatccaacaa caaatatgac atcaaatcaa 1260

tatatgattt cacatgaata tacaagttta ccaaataatt ttatgttatc aagaaatagt 1320

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1380

tggtataata attcggattg gtataataat tga 1413

<210> 64

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 64

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Asn Val Ile Ser Asp Ser Cys Asn Cys Ser

385 390 395 400

Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met

405 410 415

Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn

420 425 430

Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu

435 440 445

Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

Ser Asp Trp Tyr Asn Asn

465 470

<210> 65

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 65

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgatttcatg taattgttca 1200

acaacaaata tgacatcaaa tcaatacagt aatccaacaa caaatatgac atcaaatcaa 1260

tatatgattt cacatgaata tacaagttta ccaaataatt ttatgttatc aagaaatagt 1320

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1380

tggtataata attcggattg gtataataat tga 1413

<210> 66

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 66

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Ser Cys Asn Cys Ser

385 390 395 400

Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met

405 410 415

Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn

420 425 430

Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu

435 440 445

Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

Ser Asp Trp Tyr Asn Asn

465 470

<210> 67

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 67

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

acaacaaata tgacatcaaa tcaatacagt aatccaacaa caaatatgac atcaaatcaa 1260

tatatgattt cacatgaata tacaagttta ccaaataatt ttatgttatc aagaaatagt 1320

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1380

tggtataata attcggattg gtataataat tga 1413

<210> 68

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 68

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met

405 410 415

Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn

420 425 430

Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu

435 440 445

Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

Ser Asp Trp Tyr Asn Asn

465 470

<210> 69

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 69

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcatcaaa tcaatacagt aatccaacaa caaatatgac atcaaatcaa 1260

tatatgattt cacatgaata tacaagttta ccaaataatt ttatgttatc aagaaatagt 1320

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1380

tggtataata attcggattg gtataataat tga 1413

<210> 70

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 70

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met

405 410 415

Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn

420 425 430

Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu

435 440 445

Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

Ser Asp Trp Tyr Asn Asn

465 470

<210> 71

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 71

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca aatccaacaa caaatatgac atcaaatcaa 1260

tatatgattt cacatgaata tacaagttta ccaaataatt ttatgttatc aagaaatagt 1320

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1380

tggtataata attcggattg gtataataat tga 1413

<210> 72

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 72

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Asn Pro Thr Thr Asn Met

405 410 415

Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn

420 425 430

Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu

435 440 445

Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

Ser Asp Trp Tyr Asn Asn

465 470

<210> 73

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 73

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtatgac atcaaatcaa 1260

tatatgattt cacatgaata tacaagttta ccaaataatt ttatgttatc aagaaatagt 1320

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1380

tggtataata attcggattg gtataataat tga 1413

<210> 74

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 74

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Met

405 410 415

Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn

420 425 430

Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu

435 440 445

Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

Ser Asp Trp Tyr Asn Asn

465 470

<210> 75

<211> 1340

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 75

aatatggtcc tggtgatatg actaatggaa atcaatttat tatttcaaaa caagaatggg 60

ctactattgg agcatatatt cagactggat taggtttacc agtaaatgaa caacaattaa 120

gaacacatgt taatttaagt caggatatat caatacctag tgatttttct caattatatg 180

atgtttattg ttctgataaa acttcagcag aatggtggaa taaaaattta tatcctttaa 240

ttattaaatc tgctaatgat attgcttcat atggttttaa agttgctggt gatccttcta 300

ttaagaaaga tggatatttt aaaaaattgc aagatgaatt agataatatt gttgataata 360

attccgatga tgatgcaata gctaaagcta ttaaagattt taaagcgcga tgtggtattt 420

taattaaaga agctaaacaa tatgaagaag ctgcaaaaaa tattgtaaca tctttagatc 480

aatttttaca tggtgatcag aaaaaattag aaggtgttat caatattcaa aaacgtttaa 540

aagaagttca aacagctctt aatcaagccc atggggaaag tagtccagct cataaagagt 600

tattagaaaa agtaaaaaat ttaaaaacaa cattagaaag gactattaaa gctgaacaag 660

atttagagaa aaaagtagaa tatagttttc tattaggacc attgttagga tttgttgttt 720

atgaaattct tgaaaatact gctgttcagc atataaaaaa tcaaattgat gagataaaga 780

aacaattaga ttctgctcag catgatttgg atagagatgt taaaattata ggaatgttaa 840

atagtattaa tacagatatt gataatttat atagtcaagg acaagaagca attaaagttt 900

tccaaaagtt acaaggtatt tgggctacta ttggagctca aatagaaaat cttagaacaa 960

cgtcgttaca agaagttcaa gattctgatg atgctgatga gatacaaatt gaacttgagg 1020

acgcttctga tgcttggtta gttgtggctc aagaagctcg tgattttaca ctaaatgctt 1080

attcaactaa tagtagacaa aatttaccga ttaatgttat atcagattca tgtaattgtt 1140

caacaacaaa tatgacatca aatcaataca gtaatccaac aacaaattat atgatttcac 1200

atgaatatac aagtttacca aataatttta tgttatcaag aaatagtaat ttagaatata 1260

aatgtcctga aaataatttt atgatatatt ggtataataa ttcggattgg tataataatt 1320

cggattggta taataattga 1340

<210> 76

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 76

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn

420 425 430

Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu

435 440 445

Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

Ser Asp Trp Tyr Asn Asn

465 470

<210> 77

<211> 1413

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 77

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1320

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1380

tggtataata attcggattg gtataataat tga 1413

<210> 78

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 78

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr

420 425 430

Thr Ser Leu Pro Asn Asn Phe Met Asn Leu Glu Tyr Lys Cys Pro Glu

435 440 445

Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

Ser Asp Trp Tyr Asn Asn

465 470

<210> 79

<211> 1398

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 79

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga tcatgtaatt gttcaacaac aaatatgaca 1200

tcaaatcaat acagtaatcc aacaacaaat atgacatcaa atcaatatat gatttcacat 1260

gaatatacaa gtttaccaaa taattttatg ttatcaagaa atagtaattt agaatataaa 1320

tgtcctgaaa ataattttat gatatattgg tataataatt cggattggta taataattcg 1380

gattggtata ataattga 1398

<210> 80

<211> 465

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 80

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Ser Cys Asn Cys Ser Thr Thr Asn Met Thr

385 390 395 400

Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr

405 410 415

Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser

420 425 430

Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile

435 440 445

Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

450 455 460

Asn

465

<210> 81

<211> 1398

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 81

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcaaatcaat acagtaatcc aacaacaaat atgacatcaa atcaatatat gatttcacat 1260

gaatatacaa gtttaccaaa taattttatg ttatcaagaa atagtaattt agaatataaa 1320

tgtcctgaaa ataattttat gatatattgg tataataatt cggattggta taataattcg 1380

gattggtata ataattga 1398

<210> 82

<211> 465

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 82

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr

405 410 415

Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser

420 425 430

Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile

435 440 445

Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

450 455 460

Asn

465

<210> 83

<211> 1398

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 83

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca atgacatcaa atcaatatat gatttcacat 1260

gaatatacaa gtttaccaaa taattttatg ttatcaagaa atagtaattt agaatataaa 1320

tgtcctgaaa ataattttat gatatattgg tataataatt cggattggta taataattcg 1380

gattggtata ataattga 1398

<210> 84

<211> 465

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 84

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Met Thr Ser Asn Gln Tyr

405 410 415

Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser

420 425 430

Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile

435 440 445

Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

450 455 460

Asn

465

<210> 85

<211> 1398

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 85

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

gaatatacaa gtttaccaaa taattttatg ttatcaagaa atagtaattt agaatataaa 1320

tgtcctgaaa ataattttat gatatattgg tataataatt cggattggta taataattcg 1380

gattggtata ataattga 1398

<210> 86

<211> 465

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 86

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser

420 425 430

Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile

435 440 445

Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

450 455 460

Asn

465

<210> 87

<211> 1398

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 87

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gatttcacat ttatcaagaa atagtaattt agaatataaa 1320

tgtcctgaaa ataattttat gatatattgg tataataatt cggattggta taataattcg 1380

gattggtata ataattga 1398

<210> 88

<211> 465

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 88

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Leu Ser

420 425 430

Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile

435 440 445

Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

450 455 460

Asn

465

<210> 89

<211> 1383

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 89

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga acaacaaata tgacatcaaa tcaatacagt 1200

aatccaacaa caaatatgac atcaaatcaa tatatgattt cacatgaata tacaagttta 1260

ccaaataatt ttatgttatc aagaaatagt aatttagaat ataaatgtcc tgaaaataat 1320

tttatgatat attggtataa taattcggat tggtataata attcggattg gtataataat 1380

tga 1383

<210> 90

<211> 460

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 90

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser

385 390 395 400

Asn Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu

405 410 415

Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu

420 425 430

Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn

435 440 445

Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

<210> 91

<211> 1383

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 91

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaatgac atcaaatcaa tatatgattt cacatgaata tacaagttta 1260

ccaaataatt ttatgttatc aagaaatagt aatttagaat ataaatgtcc tgaaaataat 1320

tttatgatat attggtataa taattcggat tggtataata attcggattg gtataataat 1380

tga 1383

<210> 92

<211> 460

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 92

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu

405 410 415

Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu

420 425 430

Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn

435 440 445

Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

<210> 93

<211> 1383

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 93

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

ccaaataatt ttatgttatc aagaaatagt aatttagaat ataaatgtcc tgaaaataat 1320

tttatgatat attggtataa taattcggat tggtataata attcggattg gtataataat 1380

tga 1383

<210> 94

<211> 460

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 94

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu

420 425 430

Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn

435 440 445

Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

<210> 95

<211> 1383

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 95

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atcaatatat gattaatagt aatttagaat ataaatgtcc tgaaaataat 1320

tttatgatat attggtataa taattcggat tggtataata attcggattg gtataataat 1380

tga 1383

<210> 96

<211> 460

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 96

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Asn Ser Asn Leu

420 425 430

Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn

435 440 445

Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

450 455 460

<210> 97

<211> 1368

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 97

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga tcaaatcaat acagtaatcc aacaacaaat 1200

atgacatcaa atcaatatat gatttcacat gaatatacaa gtttaccaaa taattttatg 1260

ttatcaagaa atagtaattt agaatataaa tgtcctgaaa ataattttat gatatattgg 1320

tataataatt cggattggta taataattcg gattggtata ataattga 1368

<210> 98

<211> 455

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 98

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Ser Asn Gln Tyr Ser Asn Pro Thr Thr Asn

385 390 395 400

Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro

405 410 415

Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro

420 425 430

Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

435 440 445

Asn Ser Asp Trp Tyr Asn Asn

450 455

<210> 99

<211> 1368

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 99

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca gaatatacaa gtttaccaaa taattttatg 1260

ttatcaagaa atagtaattt agaatataaa tgtcctgaaa ataattttat gatatattgg 1320

tataataatt cggattggta taataattcg gattggtata ataattga 1368

<210> 100

<211> 455

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 100

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Glu Tyr Thr Ser Leu Pro

405 410 415

Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro

420 425 430

Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

435 440 445

Asn Ser Asp Trp Tyr Asn Asn

450 455

<210> 101

<211> 1368

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 101

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaacaaat 1260

atgacatcaa atagtaattt agaatataaa tgtcctgaaa ataattttat gatatattgg 1320

tataataatt cggattggta taataattcg gattggtata ataattga 1368

<210> 102

<211> 455

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 102

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Thr Asn Met Thr Ser Asn Ser Asn Leu Glu Tyr Lys Cys Pro

420 425 430

Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn

435 440 445

Asn Ser Asp Trp Tyr Asn Asn

450 455

<210> 103

<211> 1353

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 103

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga aatccaacaa caaatatgac atcaaatcaa 1200

tatatgattt cacatgaata tacaagttta ccaaataatt ttatgttatc aagaaatagt 1260

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1320

tggtataata attcggattg gtataataat tga 1353

<210> 104

<211> 450

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 104

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Asn Pro Thr Thr Asn Met Thr Ser Asn Gln

385 390 395 400

Tyr Met Ile Ser His Glu Tyr Thr Ser Leu Pro Asn Asn Phe Met Leu

405 410 415

Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met

420 425 430

Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr

435 440 445

Asn Asn

450

<210> 105

<211> 1353

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 105

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtttatc aagaaatagt 1260

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1320

tggtataata attcggattg gtataataat tga 1353

<210> 106

<211> 450

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 106

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Leu

405 410 415

Ser Arg Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met

420 425 430

Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr

435 440 445

Asn Asn

450

<210> 107

<211> 1353

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 107

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

tcatgtaatt gttcaacaac aaatatgaca tcaaatcaat acagtaatcc aacaaatagt 1260

aatttagaat ataaatgtcc tgaaaataat tttatgatat attggtataa taattcggat 1320

tggtataata attcggattg gtataataat tga 1353

<210> 108

<211> 450

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 108

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn

405 410 415

Pro Thr Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met

420 425 430

Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr

435 440 445

Asn Asn

450

<210> 109

<211> 1299

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 109

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga caaaatttac cgattaatgt tatatcagat 1200

aatagtaatt tagaatataa atgtcctgaa aataatttta tgatatattg gtataataat 1260

tcggattggt ataataattc ggattggtat aataattga 1299

<210> 110

<211> 432

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 110

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp

385 390 395 400

Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr

405 410 415

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

420 425 430

<210> 111

<211> 1299

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 111

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga agtttaccaa ataattttat gttatcaaga 1200

aatagtaatt tagaatataa atgtcctgaa aataatttta tgatatattg gtataataat 1260

tcggattggt ataataattc ggattggtat aataattga 1299

<210> 112

<211> 432

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 112

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Ser Leu Pro Asn Asn Phe Met Leu Ser Arg

385 390 395 400

Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr

405 410 415

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

420 425 430

<210> 113

<211> 1299

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 113

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcaac taatagtaga agtaatccaa caacaaatat gacatcaaat 1200

aatagtaatt tagaatataa atgtcctgaa aataatttta tgatatattg gtataataat 1260

tcggattggt ataataattc ggattggtat aataattga 1299

<210> 114

<211> 432

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 114

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Thr Asn Ser Arg Ser Asn Pro Thr Thr Asn Met Thr Ser Asn

385 390 395 400

Asn Ser Asn Leu Glu Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr

405 410 415

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

420 425 430

<210> 115

<211> 1251

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 115

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcagt tgctacaatt acatctggtg aaaataattt tatgatatat 1200

tggtataata attcggattg gtataataat tcggattggt ataataattg a 1251

<210> 116

<211> 416

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 116

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Val Ala Thr Ile Thr Ser Gly Glu Asn Asn Phe Met Ile Tyr

385 390 395 400

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410 415

<210> 117

<211> 1245

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 117

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcagt tgctacaatt acatctggtg aatttatgat atattggtat 1200

aataattcgg attggtataa taattcggat tggtataata attga 1245

<210> 118

<211> 414

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 118

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Val Ala Thr Ile Thr Ser Gly Glu Phe Met Ile Tyr Trp Tyr

385 390 395 400

Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410

<210> 119

<211> 1245

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 119

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttatgctac aattacatct ggtgaaaata attttatgat atattggtat 1200

aataattcgg attggtataa taattcggat tggtataata attga 1245

<210> 120

<211> 414

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 120

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ala Thr Ile Thr Ser Gly Glu Asn Asn Phe Met Ile Tyr Trp Tyr

385 390 395 400

Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410

<210> 121

<211> 1233

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 121

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatt gggttatata taatgaattt gttatgatat attggtataa taattcggat 1200

tggtataata attcggattg gtataataat tga 1233

<210> 122

<211> 410

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 122

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Trp

370 375 380

Val Ile Tyr Asn Glu Phe Val Met Ile Tyr Trp Tyr Asn Asn Ser Asp

385 390 395 400

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410

<210> 123

<211> 1251

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 123

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttattcatg ggttatatat aatgaatttg ttaataattt tatgatatat 1200

tggtataata attcggattg gtataataat tcggattggt ataataattg a 1251

<210> 124

<211> 416

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 124

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Trp Val Ile Tyr Asn Glu Phe Val Asn Asn Phe Met Ile Tyr

385 390 395 400

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410 415

<210> 125

<211> 1239

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 125

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg gatgggttat atataatgaa tttgttggta tgatatattg gtataataat 1200

tcggattggt ataataattc ggattggtat aataattga 1239

<210> 126

<211> 412

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 126

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Gly

370 375 380

Trp Val Ile Tyr Asn Glu Phe Val Gly Met Ile Tyr Trp Tyr Asn Asn

385 390 395 400

Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410

<210> 127

<211> 1251

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 127

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttatggatg ggttatatat aatgaatttg ttggtaattt tatgatatat 1200

tggtataata attcggattg gtataataat tcggattggt ataataattg a 1251

<210> 128

<211> 416

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 128

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Gly Trp Val Ile Tyr Asn Glu Phe Val Gly Asn Phe Met Ile Tyr

385 390 395 400

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410 415

<210> 129

<211> 1239

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 129

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg gagattctag tattaaaaaa gatggtttta tgatatattg gtataataat 1200

tcggattggt ataataattc ggattggtat aataattga 1239

<210> 130

<211> 412

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 130

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Gly

370 375 380

Asp Ser Ser Ile Lys Lys Asp Gly Phe Met Ile Tyr Trp Tyr Asn Asn

385 390 395 400

Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410

<210> 131

<211> 1251

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 131

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttatggaga ttctagtatt aaaaaagatg gtaataattt tatgatatat 1200

tggtataata attcggattg gtataataat tcggattggt ataataattg a 1251

<210> 132

<211> 416

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 132

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Gly Asp Ser Ser Ile Lys Lys Asp Gly Asn Asn Phe Met Ile Tyr

385 390 395 400

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410 415

<210> 133

<211> 1239

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 133

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg gagatcctag tattaaaaaa gatggtttta tgatatattg gtataataat 1200

tcggattggt ataataattc ggattggtat aataattga 1239

<210> 134

<211> 412

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 134

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Gly

370 375 380

Asp Pro Ser Ile Lys Lys Asp Gly Phe Met Ile Tyr Trp Tyr Asn Asn

385 390 395 400

Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410

<210> 135

<211> 1251

<212> DNA

<213> 人工序列

<220>

<223> 合成编码区

<400> 135

atgattattg attctaaaac aacattacca agacattctt taattcatac tataaaatta 60

aattctaata agaaatatgg tcctggtgat atgactaatg gaaatcaatt tattatttca 120

aaacaagaat gggctactat tggagcatat attcagactg gattaggttt accagtaaat 180

gaacaacaat taagaacaca tgttaattta agtcaggata tatcaatacc tagtgatttt 240

tctcaattat atgatgttta ttgttctgat aaaacttcag cagaatggtg gaataaaaat 300

ttatatcctt taattattaa atctgctaat gatattgctt catatggttt taaagttgct 360

ggtgatcctt ctattaagaa agatggatat tttaaaaaat tgcaagatga attagataat 420

attgttgata ataattccga tgatgatgca atagctaaag ctattaaaga ttttaaagcg 480

cgatgtggta ttttaattaa agaagctaaa caatatgaag aagctgcaaa aaatattgta 540

acatctttag atcaattttt acatggtgat cagaaaaaat tagaaggtgt tatcaatatt 600

caaaaacgtt taaaagaagt tcaaacagct cttaatcaag cccatgggga aagtagtcca 660

gctcataaag agttattaga aaaagtaaaa aatttaaaaa caacattaga aaggactatt 720

aaagctgaac aagatttaga gaaaaaagta gaatatagtt ttctattagg accattgtta 780

ggatttgttg tttatgaaat tcttgaaaat actgctgttc agcatataaa aaatcaaatt 840

gatgagataa agaaacaatt agattctgct cagcatgatt tggatagaga tgttaaaatt 900

ataggaatgt taaatagtat taatacagat attgataatt tatatagtca aggacaagaa 960

gcaattaaag ttttccaaaa gttacaaggt atttgggcta ctattggagc tcaaatagaa 1020

aatcttagaa caacgtcgtt acaagaagtt caagattctg atgatgctga tgagatacaa 1080

attgaacttg aggacgcttc tgatgcttgg ttagttgtgg ctcaagaagc tcgtgatttt 1140

acactaaatg cttatggaga tcctagtatt aaaaaagatg gtaataattt tatgatatat 1200

tggtataata attcggattg gtataataat tcggattggt ataataattg a 1251

<210> 136

<211> 416

<212> PRT

<213> 人工序列

<220>

<223> 从合成编码区翻译

<400> 136

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Gly Asp Pro Ser Ile Lys Lys Asp Gly Asn Asn Phe Met Ile Tyr

385 390 395 400

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410 415

<210> 137

<211> 1251

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 137

atgatcattg actccaagac caccctgccg cgtcacagcc tgatccacac catcaagctg 60

aactccaaca agaagtacgg acctggcgac atgaccaacg gcaaccagtt catcatctcc 120

aagcaagagt gggccaccat cggagcgtac atccagaccg gcctgggcct gccggtgaac 180

gagcagcagc tgaggactca cgtgaacctg tcccaagaca tcagcatccc gtccgacttc 240

tcccagctgt acgacgtgta ttgcagcgac aagacctccg ccgagtggtg gaacaagaac 300

ctgtacccgc tgatcatcaa gagcgccaac gacatcgcct cctacggctt caaggtggcg 360

ggcgacccga gcatcaagaa ggacggctac ttcaagaagc tccaagacga gctggacaac 420

atcgtggaca acaactccga cgacgacgcc atcgccaagg cgatcaagga cttcaaggcg 480

aggtgcggca tcctgatcaa ggaggcgaag cagtacgagg aggcggcgaa gaacatcgtg 540

acctccctgg accagttcct gcacggcgac cagaagaagc tggagggcgt gatcaacatc 600

cagaagaggc tgaaggaggt gcagaccgcc ctgaatcaag cccacggcga gagcagcccg 660

gcgcacaagg agctgctgga gaaggtgaag aacctcaaga ccaccctgga gaggaccatc 720

aaggccgagc aagacctgga gaagaaggtg gagtactcct tcctgctggg tccgctgctg 780

ggcttcgtgg tgtacgagat cctggagaac accgccgtgc agcacatcaa gaatcagatc 840

gacgagatca agaagcagct ggactccgcc cagcacgacc tggaccgcga cgtgaagatc 900

atcgggatgc tgaacagcat caacaccgac atcgacaacc tgtacagcca aggccaagag 960

gccatcaagg tgttccagaa gctccaaggc atctgggcga ccatcggcgc gcagatcgag 1020

aacctgagga ccaccagcct ccaagaggtc caagacagcg acgacgccga cgagatccag 1080

atcgagctgg aggacgccag cgacgcgtgg ctggtggtgg ctcaagaggc gagggacttc 1140

accctgaacg cgtacagcgt ggccaccatc accagcggcg agaacaactt catgatctac 1200

tggtacaaca actccgactg gtacaacaac tcagactggt acaacaactg a 1251

<210> 138

<211> 416

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 138

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Val Ala Thr Ile Thr Ser Gly Glu Asn Asn Phe Met Ile Tyr

385 390 395 400

Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn

405 410 415

<210> 139

<211> 1449

<212> DNA

<213> 人工序列

<220>

<223> 合成的玉蜀黍优化的编码区

<400> 139

atggctcaat ctagcagaat ctgccacggt gtgcagaacc catgtgtgat catttccaat 60

ctctccaaat ccaaccagaa caaatctcct ttctcagtca gcctcaagac tcaccagcag 120

cagcgtcgtg cttaccagat atctagctgg ggattgaaga agtcaaacaa cgggtccgtg 180

attcgtccgg ttaaggcaat gatcattgac tccaagacca ccctgccgcg tcacagcctg 240

atccacacca tcaagctgaa ctccaacaag aagtacggac ctggcgacat gaccaacggc 300

aaccagttca tcatctccaa gcaagagtgg gccaccatcg gagcgtacat ccagaccggc 360

ctgggcctgc cggtgaacga gcagcagctg aggactcacg tgaacctgtc ccaagacatc 420

agcatcccgt ccgacttctc ccagctgtac gacgtgtatt gcagcgacaa gacctccgcc 480

gagtggtgga acaagaacct gtacccgctg atcatcaaga gcgccaacga catcgcctcc 540

tacggcttca aggtggcggg cgacccgagc atcaagaagg acggctactt caagaagctc 600

caagacgagc tggacaacat cgtggacaac aactccgacg acgacgccat cgccaaggcg 660

atcaaggact tcaaggcgag gtgcggcatc ctgatcaagg aggcgaagca gtacgaggag 720

gcggcgaaga acatcgtgac ctccctggac cagttcctgc acggcgacca gaagaagctg 780

gagggcgtga tcaacatcca gaagaggctg aaggaggtgc agaccgccct gaatcaagcc 840

cacggcgaga gcagcccggc gcacaaggag ctgctggaga aggtgaagaa cctcaagacc 900

accctggaga ggaccatcaa ggccgagcaa gacctggaga agaaggtgga gtactccttc 960

ctgctgggtc cgctgctggg cttcgtggtg tacgagatcc tggagaacac cgccgtgcag 1020

cacatcaaga atcagatcga cgagatcaag aagcagctgg actccgccca gcacgacctg 1080

gaccgcgacg tgaagatcat cgggatgctg aacagcatca acaccgacat cgacaacctg 1140

tacagccaag gccaagaggc catcaaggtg ttccagaagc tccaaggcat ctgggcgacc 1200

atcggcgcgc agatcgagaa cctgaggacc accagcctcc aagaggtcca agacagcgac 1260

gacgccgacg agatccagat cgagctggag gacgccagcg acgcgtggct ggtggtggct 1320

caagaggcga gggacttcac cctgaacgcg tacagcgtgg ccaccatcac cagcggcgag 1380

aacaacttca tgatctactg gtacaacaac tccgactggt acaacaactc agactggtac 1440

aacaactga 1449

<210> 140

<211> 482

<212> PRT

<213> 人工序列

<220>

<223> 从合成的玉蜀黍优化的编码区翻译

<400> 140

Met Ala Gln Ser Ser Arg Ile Cys His Gly Val Gln Asn Pro Cys Val

1 5 10 15

Ile Ile Ser Asn Leu Ser Lys Ser Asn Gln Asn Lys Ser Pro Phe Ser

20 25 30

Val Ser Leu Lys Thr His Gln Gln Gln Arg Arg Ala Tyr Gln Ile Ser

35 40 45

Ser Trp Gly Leu Lys Lys Ser Asn Asn Gly Ser Val Ile Arg Pro Val

50 55 60

Lys Ala Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu

65 70 75 80

Ile His Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp

85 90 95

Met Thr Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr

100 105 110

Ile Gly Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln

115 120 125

Gln Leu Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser

130 135 140

Asp Phe Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala

145 150 155 160

Glu Trp Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn

165 170 175

Asp Ile Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys

180 185 190

Lys Asp Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val

195 200 205

Asp Asn Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe

210 215 220

Lys Ala Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu

225 230 235 240

Ala Ala Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp

245 250 255

Gln Lys Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu

260 265 270

Val Gln Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His

275 280 285

Lys Glu Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg

290 295 300

Thr Ile Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe

305 310 315 320

Leu Leu Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn

325 330 335

Thr Ala Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln

340 345 350

Leu Asp Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly

355 360 365

Met Leu Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly

370 375 380

Gln Glu Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr

385 390 395 400

Ile Gly Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val

405 410 415

Gln Asp Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala

420 425 430

Ser Asp Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu

435 440 445

Asn Ala Tyr Ser Val Ala Thr Ile Thr Ser Gly Glu Asn Asn Phe Met

450 455 460

Ile Tyr Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn Ser Asp Trp Tyr

465 470 475 480

Asn Asn

<210> 141

<211> 1251

<212> DNA

<213> 人工序列

<220>

<223> 玉蜀黍密码子偏倚版本DIG-1036

<400> 141

atgatcattg actccaagac caccctgccg cgtcacagcc tgatccacac catcaagctg 60

aactccaaca agaagtacgg acctggcgac atgaccaacg gcaaccagtt catcatctcc 120

aagcaagagt gggccaccat cggagcgtac atccagaccg gcctgggcct gccggtgaac 180

gagcagcagc tgaggactca cgtgcagctg tcccaagaca tcagcatccc gtccgacttc 240

tcccagctgt acgacgtgta ttgcagcgac aagacctccg ccgagtggtg gaacaagaac 300

ctgtacccgc tgatcatcaa gagcgccaac gacatcgcct cctacggctt caaggtggcg 360

ggcgacccga gcatcaagaa ggacggctac ttcaagaagc tccaagacga gctggacaac 420

atcgtggacc agaactccga cgacgacgcc atcgccaagg cgatcaagga cttcaaggcg 480

aggtgcggca tcctgatcaa ggaggcgaag cagtacgagg aggcggcgaa gaacatcgtg 540

acctccctgg accagttcct gcacggcgac cagaagaagc tggagggcgt gatcaacatc 600

cagaagaggc tgaaggaggt gcagaccgcc ctgaatcaag cccacggcga gagcagcccg 660

gcgcacaagg agctgctgga gaaggtgaag aacctcaaga ccaccctgga gaggaccatc 720

aaggccgagc aagacctgga gaagaaggtg gagtactcct tcctgctggg tccgctgctg 780

ggcttcgtgg tgtacgagat cctggagaac accgccgtgc agcacatcaa gaatcagatc 840

gacgagatca agaagcagct ggactccgcc cagcacgacc tggaccgcga cgtgaagatc 900

atcgggatgc tgaacagcat caacaccgac atcgacaacc tgtacagcca aggccaagag 960

gccatcaagg tgttccagaa gctccaaggc atctgggcga ccatcggcgc gcagatcgag 1020

aacctgagga ccaccagcct ccaagaggtc caagacagcg acgacgccga cgagatccag 1080

atcgagctgg aggacgccag cgacgcgtgg ctggtggtgg cccaagaggc gagggacttc 1140

accctgaacg cctacagcgt ggccaccatc accagcggcg agaacaactt catgatctac 1200

tggtatcaga actccgactg gtatcagaac tcagactggt acaacaactg a 1251

<210> 142

<211> 1338

<212> DNA

<213> 人工序列

<220>

<223> 最高GC+具有ER的DIG-1036的ERLS玉蜀黍最高GC版本

定位序列

<400> 142

atggcgaaca agcacctgag cctgtccctg ttcctggtgc tgcttggcct gagcgcatcc 60

ctggcatccg gaatcattga ctccaagacc accctgccgc gtcacagcct gatccacacc 120

atcaagctga actccaacaa gaagtacgga cctggcgaca tgaccaacgg caaccagttc 180

atcatctcca agcaagagtg ggccaccatc ggagcgtaca tccagaccgg cctgggcctg 240

ccggtgaacg agcagcagct gaggactcac gtgcagctgt cccaagacat cagcatcccg 300

tccgacttct cccagctgta cgacgtgtat tgcagcgaca agacctccgc cgagtggtgg 360

aacaagaacc tgtacccgct gatcatcaag agcgccaacg acatcgcctc ctacggcttc 420

aaggtggcgg gcgacccgag catcaagaag gacggctact tcaagaagct ccaagacgag 480

ctggacaaca tcgtggacca gaactccgac gacgacgcca tcgccaaggc gatcaaggac 540

ttcaaggcga ggtgcggcat cctgatcaag gaggcgaagc agtacgagga ggcggcgaag 600

aacatcgtga cctccctgga ccagttcctg cacggcgacc agaagaagct ggagggcgtg 660

atcaacatcc agaagaggct gaaggaggtg cagaccgccc tgaatcaagc ccacggcgag 720

agcagcccgg cgcacaagga gctgctggag aaggtgaaga acctcaagac caccctggag 780

aggaccatca aggccgagca agacctggag aagaaggtgg agtactcctt cctgctgggt 840

ccgctgctgg gcttcgtggt gtacgagatc ctggagaaca ccgccgtgca gcacatcaag 900

aatcagatcg acgagatcaa gaagcagctg gactccgccc agcacgacct ggaccgcgac 960

gtgaagatca tcgggatgct gaacagcatc aacaccgaca tcgacaacct gtacagccaa 1020

ggccaagagg ccatcaaggt gttccagaag ctccaaggca tctgggcgac catcggcgcg 1080

cagatcgaga acctgaggac caccagcctc caagaggtcc aagacagcga cgacgccgac 1140

gagatccaga tcgagctgga ggacgccagc gacgcgtggc tggtggtggc ccaagaggcg 1200

agggacttca ccctgaacgc ctacagcgtg gccaccatca ccagcggcga gaacaacttc 1260

atgatctact ggtatcagaa ctccgactgg tatcagaact cagactggta caacaacagc 1320

gagaaggacg agctgtga 1338

<210> 143

<211> 1347

<212> DNA

<213> 人工序列

<220>

<223> 最高GC+具有液泡的DIG-1036的VLS玉蜀黍最高GC版本

定位序列

<400> 143

atgagtagct caccgtcctt tggcctgctg gcggttgcgg ccctgcttct ggcgcttagc 60

ctggcgcaac acggcatcat tgactccaag accaccctgc cgcgtcacag cctgatccac 120

accatcaagc tgaactccaa caagaagtac ggacctggcg acatgaccaa cggcaaccag 180

ttcatcatct ccaagcaaga gtgggccacc atcggagcgt acatccagac cggcctgggc 240

ctgccggtga acgagcagca gctgaggact cacgtgcagc tgtcccaaga catcagcatc 300

ccgtccgact tctcccagct gtacgacgtg tattgcagcg acaagacctc cgccgagtgg 360

tggaacaaga acctgtaccc gctgatcatc aagagcgcca acgacatcgc ctcctacggc 420

ttcaaggtgg cgggcgaccc gagcatcaag aaggacggct acttcaagaa gctccaagac 480

gagctggaca acatcgtgga ccagaactcc gacgacgacg ccatcgccaa ggcgatcaag 540

gacttcaagg cgaggtgcgg catcctgatc aaggaggcga agcagtacga ggaggcggcg 600

aagaacatcg tgacctccct ggaccagttc ctgcacggcg accagaagaa gctggagggc 660

gtgatcaaca tccagaagag gctgaaggag gtgcagaccg ccctgaatca agcccacggc 720

gagagcagcc cggcgcacaa ggagctgctg gagaaggtga agaacctcaa gaccaccctg 780

gagaggacca tcaaggccga gcaagacctg gagaagaagg tggagtactc cttcctgctg 840

ggtccgctgc tgggcttcgt ggtgtacgag atcctggaga acaccgccgt gcagcacatc 900

aagaatcaga tcgacgagat caagaagcag ctggactccg cccagcacga cctggaccgc 960

gacgtgaaga tcatcgggat gctgaacagc atcaacaccg acatcgacaa cctgtacagc 1020

caaggccaag aggccatcaa ggtgttccag aagctccaag gcatctgggc gaccatcggc 1080

gcgcagatcg agaacctgag gaccaccagc ctccaagagg tccaagacag cgacgacgcc 1140

gacgagatcc agatcgagct ggaggacgcc agcgacgcgt ggctggtggt ggcccaagag 1200

gcgagggact tcaccctgaa cgcctacagc gtggccacca tcaccagcgg cgagaacaac 1260

ttcatgatct actggtatca gaactccgac tggtatcaga actcagactg gtacaacaac 1320

gacgagctga aggctgaggc caagtga 1347

<210> 144

<211> 416

<212> PRT

<213> 人工序列

<220>

<223>

由DIG-1036编码的多肽（DIG-1000减去糖基化

位点：N69>Q；N144>Q；N403>Q；N409>Q）

<400> 144

Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His

1 5 10 15

Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr

20 25 30

Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly

35 40 45

Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu

50 55 60

Arg Thr His Val Gln Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe

65 70 75 80

Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp

85 90 95

Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile

100 105 110

Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp

115 120 125

Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Gln

130 135 140

Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala

145 150 155 160

Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala

165 170 175

Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys

180 185 190

Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln

195 200 205

Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu

210 215 220

Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile

225 230 235 240

Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu

245 250 255

Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala

260 265 270

Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp

275 280 285

Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu

290 295 300

Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu

305 310 315 320

Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly

325 330 335

Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp

340 345 350

Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp

355 360 365

Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala

370 375 380

Tyr Ser Val Ala Thr Ile Thr Ser Gly Glu Asn Asn Phe Met Ile Tyr

385 390 395 400

Trp Tyr Gln Asn Ser Asp Trp Tyr Gln Asn Ser Asp Trp Tyr Asn Asn

405 410 415

Claims (20)

1.一种经修饰的Cry6Aa杀虫蛋白，所述经修饰的Cry6Aa杀虫蛋白包含选自由以下项组成的组的修饰：经修饰的蛋白水解易感区域，羧基末端肽(CTP)对核心蛋白的亲和力增加，以及亚细胞转运肽的添加。

2.根据权利要求1所述的经修饰的Cry6Aa杀虫蛋白，所述经修饰的Cry6Aa杀虫蛋白选自由以下项组成的组：SEQ ID NO:35、SEQ ID NO:37、SEQ ID NO:39、SEQ ID NO:41、SEQ IDNO:43、SEQ ID NO:45、SEQ ID NO:47、SEQ ID NO:49、SEQ ID NO:64、SEQ ID NO:66、SEQ IDNO:68、SEQ ID NO:70、SEQ ID NO:72、SEQ ID NO:74、SEQ ID NO:76、SEQ ID NO:78、SEQ IDNO:80、SEQ ID NO:82、SEQ ID NO:84、SEQ ID NO:86、SEQ ID NO:88、SEQ ID NO:90、SEQ IDNO:92、SEQ ID NO:94、SEQ ID NO:96、SEQ ID NO:98、SEQ ID NO:100、SEQ ID NO:102、SEQID NO:104、SEQ ID NO:106、SEQ ID NO:108、SEQ ID NO:110、SEQ ID NO:112、SEQ ID NO:114、SEQ ID NO:116、SEQ ID NO:118、SEQ ID NO:120、SEQ ID NO:132、SEQ ID NO:136、SEQID NO:138、SEQ ID NO:140和SEQ ID NO:144。

3.根据权利要求1所述的经修饰的Cry6Aa杀虫蛋白，所述经修饰的Cry6Aa杀虫蛋白选自由以下项组成的组：SEQ ID NO:110、SEQ ID NO:112、SEQ ID NO:114、SEQ ID NO:116、SEQ ID NO:118、SEQ ID NO:120、SEQ ID NO:128、SEQ ID NO:132、SEQ ID NO:140和SEQ IDNO:144。

4.根据权利要求1所述的经修饰的Cry6Aa杀虫蛋白，所述经修饰的Cry6Aa杀虫蛋白选自由以下项组成的组：SEQ ID NO:116、SEQ ID NO:118、SEQ ID NO:120、SEQ ID NO:140、SEQ ID NO:144。

5.根据权利要求1所述的经修饰的Cry6Aa杀虫蛋白，所述经修饰的Cry6Aa杀虫蛋白是SEQ ID NO:116。

6.一种编码根据权利要求1所述的经修饰的Cry6Aa杀虫蛋白的核酸序列。

7.一种编码根据权利要求2所述的经修饰的Cry6Aa杀虫蛋白的核酸序列。

8.一种编码根据权利要求3所述的经修饰的Cry6Aa杀虫蛋白的核酸序列。

9.一种编码根据权利要求4所述的经修饰的Cry6Aa杀虫蛋白的核酸序列。

10.一种编码根据权利要求5所述的经修饰的Cry6Aa杀虫蛋白的核酸序列。

11.一种包含根据权利要求6所述的核酸序列的DNA构建体，所述核酸序列与能够驱动植物中表达的启动子可操作地连接。

12.一种包含根据权利要求6所述的核酸序列的转基因植物或植物部分。

13.一种包含根据权利要求8所述的核酸序列的转基因植物或植物部分。

14.一种控制植物的害虫损伤的方法，所述方法包括传递杀虫有效量的根据权利要求1所述的杀虫蛋白质，使得害虫摄取所述杀虫蛋白质。

15.根据权利要求14所述的方法，其中所述害虫损伤是由鞘翅目昆虫引起的。

16.根据权利要求15所述的方法，其中所述害虫损伤由西方玉米根虫即玉米根萤叶甲引起。

17.一种控制对植物的害虫损伤的方法，所述方法包括制备根据权利要求12的转基因植物并将所述植物提供给害虫种群。

18.一种控制对植物的害虫损伤的方法，所述方法包括制备根据权利要求13的转基因植物并将所述植物提供给害虫种群。

19.根据权利要求18所述的方法，其中所述害虫损伤是由鞘翅目昆虫引起的。

20.根据权利要求18所述的方法，其中所述害虫损伤由西方玉米根虫即玉米根萤叶甲引起。