CN114317578A - 杀虫蛋白的用途 - Google Patents

杀虫蛋白的用途 Download PDF

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CN114317578A
CN114317578A CN202111515716.1A CN202111515716A CN114317578A CN 114317578 A CN114317578 A CN 114317578A CN 202111515716 A CN202111515716 A CN 202111515716A CN 114317578 A CN114317578 A CN 114317578A
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常藤玉
王秦扬
张爱红
陶青
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Beijing Dabeinong Biotechnology Co Ltd
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Abstract

本发明涉及一种杀虫蛋白的用途，所述杀虫蛋白可用于控制亚洲玉米螟害虫，所述控制亚洲玉米螟害虫的方法包括：将亚洲玉米螟害虫至少与ACe1蛋白接触。本发明通过细菌和/或植物体内产生能够杀死亚洲玉米螟的ACe1蛋白来控制亚洲玉米螟害虫；与现有技术使用的农业防治方法、化学防治方法、物理防治和生物防治方法相比，本发明对植物进行全生育期、全植株的保护以防治亚洲玉米螟害虫的侵害，且无污染、无残留，效果稳定、彻底，简单、方便、经济。

Description

杀虫蛋白的用途
技术领域
本发明涉及一种杀虫蛋白的用途，特别是涉及一种ACe1蛋白质通过在植物中表达来控制亚洲玉米螟为害植物的用途。
背景技术
亚洲玉米螟(Ostrinia furnacalis(Hubern))为鳞翅目螟蛾科全变态型昆虫。主要为害玉米、高粱、谷子等，也能为害棉花、甘蔗、大麻、向日葵、水稻、甜菜、甘蔗豆类等作物，属于世界性害虫。在中国所有玉米产区都有为害发生，是玉米生产上的第一大害虫。一般年份春玉米受害减产7-10％，大发生年减产30％以上。夏玉米由于生长季节短，因玉米螟为害造成的损失更大。玉米是中国最大粮食作物之一，年种植面积在6亿亩。按亩单产400公斤计算，每年因玉米螟为害而减产达240亿公斤，折合人民币约300亿元。
亚洲玉米螟一生经历卵、幼虫、蛹和成虫共4个虫态，一般一年可发生1-7代(根据纬度不同)。其幼虫期是为害玉米的时期，在玉米生产的黄淮和东北区，一年至少有两代幼虫可为害玉米。在北方春玉米区，第一代幼虫主要为害玉米苗期，幼虫一经孵化即潜入心叶丛蛀食心叶或在叶片间隙取食幼嫩叶片的叶肉造成针孔或窗户纸状的花叶，若3龄以上的幼虫蛀入未展开心叶，取食心叶造成排孔。幼虫取食心叶发育至3龄前后，此时玉米进入喇叭口期，幼嫩的雄穗对玉米螟比心叶更具有吸引力，所以幼虫将转移至雌穗取食。抽雄后幼虫被带出心叶，此时除少数老熟幼虫可在雄穗上化蛹外，所有幼虫开始向下转移，由于此时幼虫基本已是4-5龄，开始明显地发挥其“钻蛀”的特性。其中雌穗着生及其上下节是最易遭受袭击的部位。此时玉米雌穗已开始发育，如其附近茎节被蛀食，会明显地影响其正常发育甚至中止发育。此外由于玉米螟在植株中下部蛀茎，遇风极易造成倒折，使损失更大。第二代虫发生在玉米吐丝期，初孵幼虫大多集中潜藏到雌穗顶端地花丝基部内，取食花丝继而取食幼嫩籽粒，也有少数至叶腋处取食积存的花粉或叶腋组织。幼虫发育到3龄后，开始出现钻蛀行为，或直接从穗头蛀入穗轴，或继续取食正在灌浆的籽粒，或从雌穗顶向下转移再次蛀入雌穗、雌穗柄或茎秆。此时玉米雌穗己经充分发育到其应有的大小，并己进入乳熟期。因此穗期为害不会影响雌穗大小，但影响玉米正常灌浆，从而降低千粒重。如果雌穗柄被蛀空将会造成穗折而脱落。幼虫取食籽粒除直接造成产量损失外，常引发玉米穗、茎腐病的感染，更加重了产量损失和品质下降。
在中国，亚洲玉米螟的防治工作始于50年代。其所采用的主要防治方法有：农业防治、化学防治、物理防治、性信息素防治和生物防治等。
农业防治是把整个农田生态系统多因素的综合协调管理，调控作物、害虫、环境因素，创造一个有利于作物生长而不利于亚洲玉米螟发生的农田生态环境。如利用处理玉米螟越冬寄主、改革耕作制度、种植抗螟品种、设置诱集田和间作等措施减少玉米螟的为害。因农业防治必须服从作物布局和增产的要求，应用有一定的局限性，不能作为应急措施，在螟害爆发时就显得无能为力。
化学防治即农药防治，是利用化学杀虫剂来杀灭害虫，是玉米螟综合治理的重要组成部分，它具有速效、方便、简便和高经济效益的特点，特别是在害虫大发生的情况下，是必不可少的应急措施，它可以在害虫造成为害前将其消灭。如目前推广面积最大的颗粒剂防治方法，效果最为稳定和可靠，但高效施颗粒剂的工具仍远未过关，严重影响其作用的发挥。此外还有撒毒土、药液喷雾、敌敌畏封垛熏蒸秸秆垛内越冬代成虫等药剂防治方法。但化学防治也有其局限性，如使用不当往往会导致农作物发生药害、害虫产生抗药性，以及杀伤天敌，污染环境，使农田生态系统遭到破坏和农药残留对人、畜的安全构成威胁等不良后果。
物理防治主要根据害虫对环境条件中各种物理因素的反应，利用各种物理因素如光、电、色、温湿度等及机械设备进行诱杀、辐射不育等方法来防治害虫。目前应用最广泛的是高压汞灯诱杀，它利用越冬代幼虫集中在村屯中的玉米秸秆垛中越冬的特点，在越冬代成虫羽化期，大范围连片在村屯设置高压汞灯，对玉米螟成虫进行诱杀，防治效果明显。高压汞灯必须是保证夜间连续供电的村屯方能应用，且操作有一定难度。
性信息素防治是利用人工合成的玉米螟性信息素直接诱杀玉米螟雄虫和干扰螟虫交配，减少田间雌蛾受精率，减轻螟害。
其中的诱杀法是利用人工合成的玉米螟性信息素在玉米螟的交尾栖息场所如长势好的麦田或菜地等诱杀玉米螟雄蛾。此法的缺点是水盆诱捕器管理费工，并且必须大面积连片使用才有效。
其中的迷向法是在越冬代玉米螟成虫羽化10％时，在其交尾场所每公顷用性信息素散发器4500～6000个挂在作物上，干扰螟蛾的交尾。利用性信息素防治一代玉米螟，方法简便，效果良好，不污染环境，不伤害天敌。不足之处是诱芯的投放费工。
生物防治是利用某些有益生物或生物代谢产物来控制害虫种群数量，以达到降低或消灭害虫的目的。其特点是对人、畜安全，对环境污染少，对某些害虫可达到长期控制的目的。最常见的生物防治有赤眼蜂和白僵菌等。但赤眼蜂防治受气候因素影响大，效果常不稳定，并且不论螟虫发生轻重均需同样投资进行。
为了解决农业防治、化学防治、物理防治和生物防治在实际应用中的局限性，科学家们经过研究发现将编码杀虫蛋白的抗虫基因转入植物中，可获得一些抗虫转基因植物以防治植物虫害。
ACe1是一类全新杀虫蛋白，其与传统Bt蛋白完全不同。通过蛋白二级结构的分析，推测该蛋白属于β-开孔蛋白。该类蛋白的作用机制一般为酶切活化、与受体结合、形成寡聚体、在膜表面开孔。其中昆虫肠道内的酶切活化、与昆虫肠道上的受体结合以及肠道内的理化环境决定了该蛋白能否在昆虫肠道细胞膜上完成开孔。该类蛋白由菌体分泌之后，需要在作用对象体内经酶切形成活性蛋白，酶切过程主要在蛋白的氨基端或羧基端进行，将该蛋白变成活性片段。活性蛋白与昆虫肠道上皮细胞膜上的受体结合，形成寡聚体，插入肠膜，导致细胞膜出现穿孔病征，破坏细胞膜内外的渗透压变化及pH平衡等，扰乱昆虫的消化过程，最终导致其死亡。
已报道ACe1蛋白对鞘翅目玉米根虫害虫有抗虫效果。然而，至今尚无关于通过产生表达ACe1蛋白的转基因植株来控制亚洲玉米螟对植物危害的报道。
发明内容
本发明的目的是提供一种杀虫蛋白的用途，首次提供了通过产生表达ACe1蛋白来控制亚洲玉米螟的方法，且有效克服现有技术农业防治、化学防治、物理防治和生物防治等技术缺陷。
为实现上述目的，本发明提供了一种控制亚洲玉米螟害虫的方法，包括将亚洲玉米螟害虫至少与ACe1蛋白接触。
进一步地，所述ACe1蛋白存在于至少产生所述ACe1蛋白的宿主细胞中，所述亚洲玉米螟害虫通过摄食所述宿主细胞至少与所述ACe1蛋白接触。
更进一步地，所述ACe1蛋白存在于至少产生所述ACe1蛋白的细菌或转基因植物中，所述亚洲玉米螟害虫通过摄食所述细菌或所述转基因植物的组织至少与所述ACe1蛋白接触，接触后所述亚洲玉米螟害虫生长受到抑制和/或导致死亡，以实现对亚洲玉米螟危害植物的控制。
所述转基因植物可以处于任意生育期。
所述转基因植物的组织为根、叶片、茎秆、果实、雄穗、雌穗、花药或花丝。
所述对亚洲玉米螟危害植物的控制不因种植地点和/或种植时间的改变而改变。
所述植物为玉米和高粱。
所述接触步骤之前的步骤为种植含有编码所述ACe1蛋白的多核苷酸的植物。
在上述技术方案的基础上，所述ACe1蛋白为ACe1_3蛋白、ACe1_4蛋白、ACe1_5蛋白、ACe1_6蛋白、ACe1_8蛋白、ACe1_9蛋白、ACe1_10蛋白、ACe1_11蛋白、ACe1_12蛋白、ACe1_13蛋白、ACe1_14蛋白、ACe1_15蛋白、ACe1_16蛋白、ACe1_17蛋白、ACe1_18蛋白、ACe1_19蛋白、ACe1_20蛋白或ACe1_21蛋白。
优选地，所述ACe1蛋白氨基酸序列具有SEQ ID NO:1至SEQ ID NO:18所示的氨基酸序列。
在上述技术方案的基础上，所述植物还包括至少一种不同于编码所述ACe1蛋白的核苷酸的第二种核苷酸。
进一步地，所述第二种核苷酸编码Cry类杀虫蛋白质、Vip类杀虫蛋白质、蛋白酶抑制剂、凝集素、α-淀粉酶或过氧化物酶。
优选地，所述第二种核苷酸编码Cry1Ab、Cry2Ab、Cry1A.105蛋白。
更进一步地，所述Cry1Ab、Cry2Ab、Cry1A.105蛋白的氨基酸序列具有SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39所示的氨基酸序列。所述第二种核苷酸具有SEQ ID NO:40、SEQ ID NO:41、SEQ ID NO:42所示的核苷酸序列。
可选择地，所述第二种核苷酸为抑制目标昆虫害虫中重要基因的dsRNA。
为实现上述目的，本发明还提供了一种ACe1蛋白质控制亚洲玉米螟害虫的用途。
为实现上述目的，本发明还提供了一种产生控制亚洲玉米螟害虫的植物的方法，包括向所述植物的基因组中引入编码ACe1蛋白的多核苷酸序列。
为实现上述目的，本发明还提供了一种产生控制亚洲玉米螟害虫的植物种子的方法，包括将由所述方法获得的第一植株与第二植株杂交，从而产生含有编码ACe1蛋白的多核苷酸序列的种子。
为实现上述目的，本发明还提供了一种培养控制亚洲玉米螟害虫的植物的方法，包括：
种植至少一粒植物种子，所述植物种子的基因组中包括编码ACe1蛋白的多核苷酸序列；
使所述植物种子长成植株；
使所述植株在人工接种亚洲玉米螟害虫和/或亚洲玉米螟害虫自然发生危害的条件下生长，收获与其他不具有编码ACe1蛋白的多核苷酸序列的植株相比具有减弱的植物损伤和/或具有增加的植物产量的植株。
本发明中所述的“接触”，是指昆虫和/或害虫触碰、停留和/或摄食植物、植物器官、植物组织或植物细胞，所述植物、植物器官、植物组织或植物细胞既可以是其体内表达杀虫蛋白，还可以是所述植物、植物器官、植物组织或植物细胞的表面具有杀虫蛋白和/或具有产生杀虫蛋白的微生物。
本发明术语“控制”和/或“防治”是指亚洲玉米螟害虫至少与ACe1蛋白接触，接触后亚洲玉米螟害虫生长受到抑制和/或导致死亡。进一步地，亚洲玉米螟害虫通过摄食植物组织至少与ACe1蛋白接触，接触后全部或部分亚洲玉米螟害虫生长受到抑制和/或导致死亡。抑制是指亚致死，即尚未致死但能引起生长发育、行为、生理、生化和组织等方面的某种效应，如生长发育缓慢和/或停止。同时，植物在形态上应是正常的，且可在常规方法下培养以用于产物的消耗和/或生成。此外，含有编码ACe1蛋白的多核苷酸序列的控制亚洲玉米螟害虫的植物和/或植物种子，在人工接种亚洲玉米螟害虫和/或亚洲玉米螟害虫自然发生危害的条件下，与非转基因的野生型植株相比具有减弱的植物损伤，具体表现包括但不限于改善的茎秆抗性、和/或提高的籽粒重量、和/或增产等。ACe1蛋白对亚洲玉米螟的“控制”和/或“防治”作用是可以独立存在的，不因其它可“控制”和/或“防治”亚洲玉米螟害虫的物质的存在而减弱和/或消失。具体地，转基因植物(含有编码ACe1蛋白的多核苷酸序列)的任何组织同时和/或不同步地，存在和/或产生，ACe1蛋白和/或可控制亚洲玉米螟害虫的另一种物质，则所述另一种物质的存在既不影响ACe1蛋白对亚洲玉米螟的“控制”和/或“防治”作用，也不能导致所述“控制”和/或“防治”作用完全和/或部分由所述另一种物质实现，而与ACe1蛋白无关。通常情况下，在大田，亚洲玉米螟害虫摄食植物组织的过程短暂且很难用肉眼观察到，因此，在人工接种亚洲玉米螟害虫和/或亚洲玉米螟害虫自然发生危害的条件下，如转基因植物(含有编码ACe1蛋白的多核苷酸序列)的任何组织存在死亡的亚洲玉米螟、和/或在其上停留生长受到抑制的亚洲玉米螟害虫、和/或与非转基因的野生型植株相比具有减弱的植物损伤，即为实现了本发明的方法和/或用途，即通过亚洲玉米螟害虫至少与ACe1蛋白接触以实现控制亚洲玉米螟害虫的方法和/或用途。
在本发明中，ACe1蛋白在一种转基因植物中的表达可以伴随着一个或多个Cry类杀虫蛋白质和/或Vip类杀虫蛋白质的表达。这种超过一种的杀虫毒素在同一株转基因植物中共同表达可以通过遗传工程使植物包含并表达所需的基因来实现。另外，一种植物(第1亲本)可以通过遗传工程操作表达ACe1蛋白质，第二种植物(第2亲本)可以通过遗传工程操作表达Cry类杀虫蛋白质和/或Vip类杀虫蛋白质。通过第1亲本和第2亲本杂交获得表达引入第1亲本和第2亲本的所有基因的后代植物。
RNA干扰(RNA interference，RNAi)是指在进化过程中高度保守的、由双链RNA(double-stranded RNA，dsRNA)诱发的、同源mRNA高效特异性降解的现象。因此在本发明中可以使用RNAi技术特异性剔除或关闭目标昆虫害虫中特定基因的表达。
本发明所述的亚洲玉米螟(Ostrinia furnacalis(Hubern))为我国玉米生产中为害最广泛也最严重的害虫，其属于鳞翅目螟蛾科全变态型昆虫。雄成虫体长13-14mm，翅展22-28mm，体背黄褐色，前翅内横线为黄褐色波浪纹，外横线暗褐色，呈锯齿状纹。雌成虫体长约14-15毫米，翅展28-34毫米，体鲜黄色，各条线纹红褐色。老熟幼虫，体长20-30毫米，圆筒形，头黑褐色，背部淡灰色或略带淡红褐色幼虫中、后胸背面各有1排4个圆形毛片，腹部1～8节背面前方有1排4个圆形毛片，后方两个，较前排稍小。
亚洲玉米螟为我国分布最广，为害最严重的玉米害虫，根据中国农技推广中心的统计，其每年为害面积可达3.5亿亩，分布于东北、黄淮以及西南等各地。普遍可造成5-10％的损失，严重地区可造成30％的损失。
本发明所述的小地老虎(Agrotis ypsilon(Rottemberg))是一种常见农业害虫，属于鳞翅目夜蛾科杂食性昆虫。主要为害玉米，大豆，棉花，油菜等。以幼虫取食作物茎基部，造成倒伏或死亡，严重时可造成缺苗断垄。
本发明所述的东方黏虫(Mythimna separate(Walker))是鳞翅目夜蛾科迁飞性杂食性害虫，为害玉米、水稻、高粱等作物。东方黏虫喜食叶，3龄后能将整片叶咬食成缺刻状，或吃光心叶，形成无心苗；5至6龄达暴食期，能将幼苗地上部分全部吃光，或将整个植株叶片吃掉只留下叶脉，造成严重减产，甚至绝收。
本发明中所述的ACe1蛋白是一类β-开孔蛋白，而昆虫肠道内的酶切活化、与昆虫肠道上的受体结合以及肠道内的理化环境是β-开孔蛋白起作用的关键点，只有能够将β-开孔蛋白进行酶切成活性片段后，并与昆虫肠道上皮细胞膜上的受体结合，才有可能使得某个β-开孔蛋白对该害虫具有抗虫效果。受体结合过程需要精确匹配，往往开孔蛋白或受体蛋白上的一个氨基酸差异就能造成与同一受体的结合发生改变。例如同属于β-开孔蛋白的气溶素蛋白(aerolysin)在R336A突变后，对CTLL-2细胞系的毒力产生了质的变化(Osusky,Teschk等，2008)。同样由于受体发生了变化也可导致同一个β-开孔蛋白的毒力发生变化。例如采用dsRNA将MDCK细胞系上的HAVCR1基因进行抑制，导致了ε-毒蛋白(epsilon-toxin)对细胞的毒力产生了百倍差异(Ivie,Fennessey等，2011)。这充分说明了β-开孔蛋白与昆虫体内酶和受体的相互作用方式是复杂且难以预料的。
本发明中所述的植物、植物组织或植物细胞的基因组，是指植物、植物组织或植物细胞内的任何遗传物质，且包括细胞核和质体和线粒体基因组。
本发明中所述的多核苷酸和/或核苷酸形成完整“基因”，在所需宿主细胞中编码蛋白质或多肽。本领域技术人员很容易认识到，可以将本发明的多核苷酸和/或核苷酸置于目的宿主中的调控序列控制下。
本领域技术人员所熟知的，DNA典型的以双链形式存在。在这种排列中，一条链与另一条链互补，反之亦然。由于DNA在植物中复制产生了DNA的其它互补链。这样，本发明包括对序列表中示例的多核苷酸及其互补链的使用。本领域常使用的“编码链”指与反义链结合的链。为了在体内表达蛋白质，典型将DNA的一条链转录为一条mRNA的互补链，它作为模板翻译出蛋白质。mRNA实际上是从DNA的“反义”链转录的。“有义”或“编码”链有一系列密码子(密码子是三个核苷酸，一次读三个可以产生特定氨基酸)，其可作为开放阅读框(ORF)阅读来形成目的蛋白质或肽。本发明还包括与示例的DNA有相当功能的RNA。
本发明中核酸分子或其片段在严格条件下与本发明ACe1基因杂交。任何常规的核酸杂交或扩增方法都可以用于鉴定本发明ACe1基因的存在。核酸分子或其片段在一定情况下能够与其他核酸分子进行特异性杂交。本发明中，如果两个核酸分子能形成反平行的双链核酸结构，就可以说这两个核酸分子彼此间能够进行特异性杂交。如果两个核酸分子显示出完全的互补性，则称其中一个核酸分子是另一个核酸分子的“互补物”。本发明中，当一个核酸分子的每一个核苷酸都与另一个核酸分子的对应核苷酸互补时，则称这两个核酸分子显示出“完全互补性”。如果两个核酸分子能够以足够的稳定性相互杂交从而使它们在至少常规的“低度严格”条件下退火且彼此结合，则称这两个核酸分子为“最低程度互补”。类似地，如果两个核酸分子能够以足够的稳定性相互杂交从而使它们在常规的“高度严格”条件下退火且彼此结合，则称这两个核酸分子具有“互补性”。从完全互补性中偏离是可以允许的，只要这种偏离不完全阻止两个分子形成双链结构。为了使一个核酸分子能够作为引物或探针，仅需保证其在序列上具有充分的互补性，以使得在所采用的特定溶剂和盐浓度下能形成稳定的双链结构。
本发明中，基本同源的序列是一段核酸分子，该核酸分子在高度严格条件下能够和相匹配的另一段核酸分子的互补链发生特异性杂交。促进DNA杂交的适合的严格条件，例如，大约在45℃条件下用6.0×氯化钠/柠檬酸钠(SSC)处理，然后在50℃条件下用2.0×SSC洗涤，这些条件对本领域技术人员是公知的。例如，在洗涤步骤中的盐浓度可以选自低度严格条件的约2.0×SSC、50℃到高度严格条件的约0.2×SSC、50℃。此外，洗涤步骤中的温度条件可以从低度严格条件的室温约22℃，升高到高度严格条件的约65℃。温度条件和盐浓度可以都发生改变，也可以其中一个保持不变而另一个变量发生改变。优选地，本发明所述严格条件可为在6×SSC、0.5％SDS溶液中，在65℃下发生特异性杂交，然后用2×SSC、0.1％SDS和1×SSC、0.1％SDS各洗膜1次。
因此，具有抗虫活性并在严格条件下与本发明SEQ ID NO:19至SEQ ID NO:36杂交的序列包括在本发明中。这些序列与本发明序列至少大约40％-50％同源，大约60％、65％或70％同源，甚至至少大约75％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更大的序列同源性。
本发明中所述的基因和蛋白质不但包括特定的示例序列，还包括保存了所述特定示例的蛋白质的杀虫活性特征的部分和/片段(包括与全长蛋白质相比在内和/或末端缺失)、变体、突变体、取代物(有替代氨基酸的蛋白质)、嵌合体和融合蛋白。所述“变体”或“变异”是指编码同一蛋白或编码有杀虫活性的等价蛋白的核苷酸序列。所述“等价蛋白”是指与权利要求的蛋白具有相同或基本相同的抗亚洲玉米螟害虫的生物活性的蛋白。
本发明中所述的DNA分子或蛋白序列的“片段”或“截短”是指涉及的原始DNA或蛋白序列(核苷酸或氨基酸)的一部分或其人工改造形式(例如适合植物表达的序列)，前述序列的长度可存在变化，但长度足以确保(编码)蛋白质为昆虫毒素。
使用标准技术可以修饰基因和容易的构建基因变异体。例如，本领域熟知制造点突变的技术。又例如美国专利号5605793描述了在随机断裂后使用DNA重装配产生其它分子多样性的方法。可以使用商业化核酸内切酶制造全长基因的片段，并且可以按照标准程序使用核酸外切酶。例如，可以使用酶诸如Bal31或定点诱变从这些基因的末端系统地切除核苷酸。还可以使用多种限制性内切酶获取编码活性片段的基因。可以使用蛋白酶直接获得这些毒素的活性片段。
本发明可以从β-开孔蛋白分离物和/或DNA文库衍生出等价蛋白和/或编码这些等价蛋白的基因。有多种方法获取本发明的杀虫蛋白。例如，可以使用本发明公开和要求保护的杀虫蛋白的抗体从蛋白质混合物鉴定和分离其它蛋白。特别地，抗体可能是由蛋白最恒定和与其它β-开孔蛋白最不同的蛋白部分引起的。然后可以通过免疫沉淀、酶联免疫吸附测定(ELISA)或western印迹方法使用这些抗体专一地鉴定有特征活性的等价蛋白。可使用本领域标准程序容易的制备本发明中公开的蛋白或等价蛋白或这类蛋白的片段的抗体。然后可以从微生物中获得编码这些蛋白的基因。
由于遗传密码子的丰余性，多种不同的DNA序列可以编码相同的氨基酸序列。产生这些编码相同或基本相同的蛋白的可替代DNA序列正在本领域技术人员的技术水平内。这些不同的DNA序列包括在本发明的范围内。所述“基本上相同的”序列是指有氨基酸取代、缺失、添加或插入但实质上不影响杀虫活性的序列，亦包括保留杀虫活性的片段。
本发明中氨基酸序列的取代、缺失或添加是本领域的常规技术，优选这种氨基酸变化为：小的特性改变，即不显著影响蛋白的折叠和/或活性的保守氨基酸取代；小的缺失，通常约1-30个氨基酸的缺失；小的氨基或羧基端延伸，例如氨基端延伸一个甲硫氨酸残基；小的连接肽，例如约20-25个残基长。
保守取代的实例是在下列氨基酸组内发生的取代：碱性氨基酸(如精氨酸、赖氨酸和组氨酸)、酸性氨基酸(如谷氨酸和天冬氨酸)、极性氨基酸(如谷氨酰胺、天冬酰胺)、疏水性氨基酸(如亮氨酸、异亮氨酸和缬氨酸)、芳香氨基酸(如苯丙氨酸、色氨酸和酪氨酸)，以及小分子氨基酸(如甘氨酸、丙氨酸、丝氨酸、苏氨酸和甲硫氨酸)。通常不改变特定活性的那些氨基酸取代在本领域内是众所周知的，并且已由，例如，N.Neurath和R.L.Hill在1979年纽约学术出版社(Academic Press)出版的《Protein》中进行了描述。最常见的互换有Ala/Ser，Val/Ile，Asp/Glu，Thu/Ser，Ala/Thr，Ser/Asn，Ala/Val，Ser/Gly，Tyr/Phe，Ala/Pro，Lys/Arg，Asp/Asn，Leu/Ile，Leu/Val，Ala/Glu和Asp/Gly，以及它们相反的互换。
对于本领域的技术人员而言显而易见地，这种取代可以在对分子功能起重要作用的区域之外发生，而且仍产生活性多肽。对于由本发明的多肽，其活性必需的并因此选择不被取代的氨基酸残基，可以根据本领域已知的方法，如定点诱变或丙氨酸扫描诱变进行鉴定(如参见，Cunningham和Wells，1989，Science 244：1081-1085)。后一技术是在分子中每一个带正电荷的残基处引入突变，检测所得突变分子的抗虫活性，从而确定对该分子活性而言重要的氨基酸残基。底物-酶相互作用位点也可以通过其三维结构的分析来测定，这种三维结构可由核磁共振分析、结晶学或光亲和标记等技术测定(参见，如de Vos等，1992，Science 255：306-312；Smith等，1992，J.Mol.Biol 224:899-904；Wlodaver等，1992，FEBSLetters 309：59-64)。
在本发明中，ACe1蛋白包括但不限于SEQ ID NO:1至SEQ ID NO:18，与SEQ ID NO:1至SEQ ID NO:18所示的氨基酸序列具有一定同源性的氨基酸序列也包括在本发明中。这些序列与本发明序列类似性/相同性典型的大于78％，优选的大于85％，更优选的大于90％，甚至更优选的大于95％，并且可以大于99％。也可以根据更特定的相同性和/或类似性范围定义本发明的优选的多核苷酸和蛋白质。例如与本发明示例的序列有78％、79％、80％、81％、82％、83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％的相同性和/或类似性。
本发明中所述调控序列包括但不限于启动子、转运肽、终止子、增强子、前导序列、内含子以及其它可操作地连接到所述ACe1蛋白的调节序列。
所述启动子为植物中可表达的启动子，所述的“植物中可表达的启动子”是指确保与其连接的编码序列在植物细胞内进行表达的启动子。植物中可表达的启动子可为组成型启动子。指导植物内组成型表达的启动子的示例包括但不限于，来源于花椰菜花叶病毒的35S启动子、玉米Ubi启动子、水稻GOS2基因的启动子等。备选地，植物中可表达的启动子可为组织特异的启动子，即该启动子在植物的一些组织内如在绿色组织中指导编码序列的表达水平高于植物的其他组织(可通过常规RNA试验进行测定)，如PEP羧化酶启动子。备选地，植物中可表达的启动子可为创伤诱导启动子。创伤诱导启动子或指导创伤诱导的表达模式的启动子是指当植物经受机械或由昆虫啃食引起的创伤时，启动子调控下的编码序列的表达较正常生长条件下有显著提高。创伤诱导启动子的示例包括但不限于，马铃薯和西红柿的蛋白酶抑制基因(pinⅠ和pinⅡ)和玉米蛋白酶抑制基因(MPI)的启动子。
所述转运肽(又称分泌信号序列或导向序列)是指导转基因产物到特定的细胞器或细胞区室，对受体蛋白质来说，所述转运肽可以是异源的，例如，利用编码叶绿体转运肽序列靶向叶绿体，或者利用‘KDEL’保留序列靶向内质网，或者利用大麦植物凝集素基因的CTPP靶向液泡。
所述前导序列包含但不限于，小RNA病毒前导序列，如EMCV前导序列(脑心肌炎病毒5’非编码区)；马铃薯Y病毒组前导序列，如MDMV(玉米矮缩花叶病毒)前导序列；人类免疫球蛋白质重链结合蛋白质(BiP)；苜蓿花叶病毒的外壳蛋白质mRNA的不翻译前导序列(AMVRNA4)；烟草花叶病毒(TMV)前导序列。
所述增强子包含但不限于，花椰菜花叶病毒(CaMV)增强子、玄参花叶病毒(FMV)增强子、康乃馨风化环病毒(CERV)增强子、木薯脉花叶病毒(CsVMV)增强子、紫茉莉花叶病毒(MMV)增强子、夜香树黄化曲叶病毒(CmYLCV)增强子、木尔坦棉花曲叶病毒(CLCuMV)、鸭跖草黄斑驳病毒(CoYMV)和花生褪绿线条花叶病毒(PCLSV)增强子。
对于单子叶植物应用而言，所述内含子包含但不限于，玉米hsp70内含子、玉米泛素内含子、Adh内含子1、蔗糖合酶内含子或水稻Act1内含子。对于双子叶植物应用而言，所述内含子包含但不限于，CAT-1内含子、pKANNIBAL内含子、PIV2内含子和“超级泛素”内含子。
所述终止子可以为在植物中起作用的适合多聚腺苷酸化信号序列，包括但不限于，来源于农杆菌(Agrobacterium tumefaciens)胭脂碱合成酶(NOS)基因的多聚腺苷酸化信号序列、来源于蛋白酶抑制剂Ⅱ(pinⅡ)基因的多聚腺苷酸化信号序列、来源于豌豆ssRUBISCO E9基因的多聚腺苷酸化信号序列和来源于α-微管蛋白(α-tubulin)基因的多聚腺苷酸化信号序列。
本发明中所述“有效连接”表示核酸序列的联结，所述联结使得一条序列可提供对相连序列来说需要的功能。在本发明中所述“有效连接”可以为将启动子与感兴趣的序列相连，使得该感兴趣的序列的转录受到该启动子控制和调控。当感兴趣的序列编码蛋白并且想要获得该蛋白的表达时“有效连接”表示：启动子与所述序列相连，相连的方式使得得到的转录物高效翻译。如果启动子与编码序列的连接是转录物融合并且想要实现编码的蛋白的表达时，制造这样的连接，使得得到的转录物中第一翻译起始密码子是编码序列的起始密码子。备选地，如果启动子与编码序列的连接是翻译融合并且想要实现编码的蛋白的表达时，制造这样的连接，使得5’非翻译序列中含有的第一翻译起始密码子与启动子相连结，并且连接方式使得得到的翻译产物与编码想要的蛋白的翻译开放读码框的关系是符合读码框的。可以“有效连接”的核酸序列包括但不限于：提供基因表达功能的序列(即基因表达元件，例如启动子、5’非翻译区域、内含子、蛋白编码区域、3’非翻译区域、聚腺苷化位点和/或转录终止子)、提供DNA转移和/或整合功能的序列(即T-DNA边界序列、位点特异性重组酶识别位点、整合酶识别位点)、提供选择性功能的序列(即抗生素抗性标记物、生物合成基因)、提供可计分标记物功能的序列、体外或体内协助序列操作的序列(即多接头序列、位点特异性重组序列)和提供复制功能的序列(即细菌的复制起点、自主复制序列、着丝粒序列)。
本发明中所述的“杀虫”或“抗虫”是指对农作物害虫是有毒的，从而实现“控制”和/或“防治”农作物害虫。优选地，所述“杀虫”或“抗虫”是指杀死农作物害虫。更具体地，目标昆虫是亚洲玉米螟害虫。
本发明中ACe1蛋白对亚洲玉米螟害虫具有毒性。本发明中的植物，特别是玉米，在其基因组中含有外源DNA，所述外源DNA包含编码ACe1蛋白的核苷酸序列，亚洲玉米螟害虫通过摄食植物组织与该蛋白接触，接触后亚洲玉米螟害虫生长受到抑制和/或导致死亡。抑制是指致死或亚致死。同时，植物在形态上应是正常的，且可在常规方法下培养以用于产物的消耗和/或生成。此外，该植物可基本消除对化学或生物杀虫剂的需要(所述化学或生物杀虫剂为针对ACe1蛋白所靶向的亚洲玉米螟害虫的杀虫剂)。
植物材料中杀虫蛋白的表达水平可通过本领域内所描述的多种方法进行检测，例如通过应用特异引物对组织内产生的编码杀虫蛋白质的mRNA进行定量，或直接特异性检测产生的杀虫蛋白质的量。
可以应用不同的试验测定植物中杀虫蛋白的杀虫效果。本发明中目标昆虫主要为亚洲玉米螟。
本发明中，所述ACe1蛋白可以具有序列表中SEQ ID NO:1至SEQ ID NO:18所示的氨基酸序列。除了包含ACe1蛋白的编码区外，也可包含其他元件，例如编码选择性标记的蛋白质。
此外，包含编码本发明ACe1蛋白的核苷酸序列的表达盒在植物中还可以与至少一种编码除草剂抗性基因的蛋白质一起表达，所述除草剂抗性基因包括但不限于，草胺膦抗性基因(如bar基因、pat基因)、苯敌草抗性基因(如pmph基因)、草甘膦抗性基因(如EPSPS基因)、溴苯腈(bromoxynil)抗性基因、磺酰脲抗性基因、对除草剂茅草枯的抗性基因、对氨腈的抗性基因或谷氨酰胺合成酶抑制剂(如PPT)的抗性基因，从而获得既具有高杀虫活性、又具有除草剂抗性的转基因植物。
本发明中，将外源DNA导入植物，如将编码所述ACe1蛋白的基因或表达盒或重组载体导入植物细胞，常规的转化方法包括但不限于，农杆菌介导的转化、微量发射轰击、直接将DNA摄入原生质体、电穿孔或晶须硅介导的DNA导入。
本发明提供了一种杀虫蛋白的用途，具有以下优点：
1、内因防治。现有技术主要是通过外部作用即外因来控制亚洲玉米螟害虫的危害，如农业防治、化学防治、物理防治和生物防治；而本发明是通过植物体内产生能够杀死亚洲玉米螟的ACe1蛋白来控制亚洲玉米螟害虫的，即通过内因来防治。
2、无污染、无残留。现有技术使用的化学防治方法虽然对控制亚洲玉米螟害虫的危害起到了一定作用，但同时也对人、畜和农田生态系统带来了污染、破坏和残留；使用本发明控制亚洲玉米螟害虫的方法，可以消除上述不良后果。
3、全生育期防治。现有技术使用的控制亚洲玉米螟害虫的方法都是阶段性的，而本发明是对植物进行全生育期的保护，转基因植物(ACe1蛋白)从发芽、生长，一直到开花、结果，都可以避免遭受亚洲玉米螟的侵害。
4、全植株防治。现有技术使用的控制亚洲玉米螟害虫的方法大多是局部性的，如叶面喷施；而本发明是对整个植株进行保护，如转基因植物(ACe1蛋白)的根、叶片、茎秆、果实、雄穗、雌穗、花药或花丝等都是可以抵抗亚洲玉米螟侵害的。
5、效果稳定。现有技术使用的无论是农业防治方法还是物理防治方法都需要利用环境条件对害虫进行防治，可变因素较多；本发明是使所述ACe1蛋白在植物体内进行表达，有效地克服了环境条件不稳定的缺陷，且本发明转基因植物(ACe1蛋白)的防治效果在不同地点、不同时间、不同遗传背景也都是稳定一致的。
6、简单、方便、经济。本发明只需种植能够表达ACe1蛋白的转基因植物即可，而不需要采用其它措施，从而节省了大量人力、物力和财力。
7、效果彻底。现有技术使用的控制亚洲玉米螟害虫的方法，其效果是不彻底的，只起到减轻作用；而本发明转基因植物(ACe1蛋白)可以造成初孵亚洲玉米螟幼虫的大量死亡。
下面通过附图和实施例，对本发明的技术方案做进一步的详细描述。
附图说明
图1为本发明杀虫蛋白的用途的含有ACe1核苷酸序列的重组表达载体DBN01-P构建流程图。
具体实施方式
下面通过具体实施例进一步说明本发明杀虫蛋白的用途的技术方案。
第一实施例、基因的获得和合成
1、获得核苷酸序列
ACe1杀虫蛋白的氨基酸序列，如表1所示；编码相应于所述ACe1杀虫蛋白的氨基酸序列的ACe1核苷酸序列，如表1所示。
表1、ACe1蛋白及其对应的氨基酸及核苷酸序列
杀虫蛋白名称 氨基酸序列号 细菌核苷酸序列号 植物核苷酸序列号
ACe1_3 SEQ ID NO:1 SEQ ID NO:19 SEQ ID NO:43
ACe1_4 SEQ ID NO:2 SEQ ID NO:20 SEQ ID NO:44
ACe1_5 SEQ ID NO:3 SEQ ID NO:21 SEQ ID NO:45
ACe1_6 SEQ ID NO:4 SEQ ID NO:22 SEQ ID NO:46
ACe1_8 SEQ ID NO:5 SEQ ID NO:23 SEQ ID NO:47
ACe1_9 SEQ ID NO:6 SEQ ID NO:24 SEQ ID NO:48
ACe1_10 SEQ ID NO:7 SEQ ID NO:25 SEQ ID NO:49
ACe1_11 SEQ ID NO:8 SEQ ID NO:26 SEQ ID NO:50
ACe1_12 SEQ ID NO:9 SEQ ID NO:27 SEQ ID NO:51
ACe1_13 SEQ ID NO:10 SEQ ID NO:28 SEQ ID NO:52
ACe1_14 SEQ ID NO:11 SEQ ID NO:29 SEQ ID NO:53
ACe1_15 SEQ ID NO:12 SEQ ID NO:30 SEQ ID NO:54
ACe1_16 SEQ ID NO:13 SEQ ID NO:31 SEQ ID NO:55
ACe1_17 SEQ ID NO:14 SEQ ID NO:32 SEQ ID NO:56
ACe1_18 SEQ ID NO:15 SEQ ID NO:33 SEQ ID NO:57
ACe1_19 SEQ ID NO:16 SEQ ID NO:34 SEQ ID NO:58
ACe1_20 SEQ ID NO:17 SEQ ID NO:35 SEQ ID NO:59
ACe1_21 SEQ ID NO:18 SEQ ID NO:36 SEQ ID NO:60
2、合成上述核苷酸序列
上述18条ACe1蛋白的核苷酸序列(如序列表SEQ ID NO:19至SEQ ID NO:36)由南京金斯瑞生物科技有限公司分别合成。
第二实施例、重组表达载体的构建和重组表达载体转化大肠杆菌获得ACe1蛋白
1、构建含有ACe1基因的重组表达载体
将第一实施例合成的ACe1蛋白(ACe1_3至ACe1_6、ACe1_8至ACe1_21)的核苷酸序列连入蛋白表达载体pET28a(Novagen，USA，CAT：69864-3)上，操作步骤按Novagen公司产品pET28a载体说明书进行，得到重组表达载体DBN01-P至DBN18-P，其构建流程如图1所示(其中，Kan表示卡纳霉素抗性基因；f1 ori表示噬菌体f1的复制起点；Lacl为Lacl起始密码子；ACe1_3为ACe1_3核苷酸序列(SEQ ID NO:19)；MCS为多克隆位点)。
ACe1蛋白及其对应的重组表达载体名称，如表2所示：
表2、ACe1蛋白及其对应的重组表达载体名称
杀虫蛋白名称 重组表达载体
ACe1_3 DBN01-P
ACe1_4 DBN02-P
ACe1_5 DBN03-P
ACe1_6 DBN04-P
ACe1_8 DBN05-P
ACe1_9 DBN06-P
ACe1_10 DBN07-P
ACe1_11 DBN08-P
ACe1_12 DBN09-P
ACe1_13 DBN10-P
ACe1_14 DBN11-P
ACe1_15 DBN12-P
ACe1_16 DBN13-P
ACe1_17 DBN14-P
ACe1_18 DBN15-P
ACe1_19 DBN16-P
ACe1_20 DBN17-P
ACe1_21 DBN18-P
2、重组表达载体转化大肠杆菌获得ACe1蛋白
然后将重组表达载体DBN01-P至DBN18-P用热激方法转化大肠杆菌BL21(DE3)感受态细胞(Transgen，China，CAT：CD501)，挑取阳性克隆于LB液体培养基(胰蛋白胨10g/L、酵母提取物5g/L、NaCl 10g/L、氨苄青霉素100mg/L，用NaOH调pH至7.5)中于温度37℃，转速200r/min条件下培养16h。再将培养液按照1：10的比例转接到YT培养基中，置于温度37℃，转速200r/min条件下培养。当培养液的OD＝600值达到0.6-0.8时，加入IPTG至终浓度为0.5mM进行诱导表达6h，离心培养液收集菌体，弃上清，加入PBS重悬后超声波破碎，用SDS-PAGE对表达蛋白进行检测，估算蛋白浓度，于温度-20℃保存备用。
第三实施例、饲喂ACe1蛋白鉴定对亚洲玉米螟的抗虫效果
将实施例二中所述的2获得的ACe1系列(ACe1_3至ACe1_6、ACe1_8至ACe1_21)蛋白分别对玉米螟、小地老虎、东方黏虫进行抗虫效果检测。每一个虫子共设置18组处理，分别为：ACe1_3至ACe1_6、ACe1_8至ACe1_21，以及1组阴性对照处理：GFP。将ACe1_3至ACe1_6、ACe1_8至ACe1_21、GFP的蛋白液分别混合到饲料中，终浓度为50μg/g。每组处理进行3次重复。
表3、ACe1蛋白饲喂玉米螟、小地老虎、东方黏虫的抗虫结果
Figure BDA0003406773390000151
“+”代表有致死效果；“-”代表无抗虫效果；“NT”代表未测试；“S”代表发育抑制
表3的结果表明：ACe1_3至ACe1_6、ACe1_8至ACe1_11、ACe1_13、ACe1_14、ACe1_16至ACe1_18蛋白均对玉米螟表现出良好的抗虫效果，而对小地老虎和东方黏虫仅表现出抑制发育的效果。
由此证明ACe1蛋白(ACe1_3至ACe1_6、ACe1_8至ACe1_11、ACe1_13、ACe1_14、ACe1_16至ACe1_18)都显示出对玉米螟的抗性活性，这种活性足以对玉米螟的生长产生不良效应从而使其在田间得以控制。同时通过控制玉米螟为害，也有可能降低转ACe1基因植株上病害的发生，极大的提高转ACe1基因植株的产量及品质。
综上所述，本发明杀虫蛋白的用途通过植物体内产生能够杀死玉米螟的ACe1蛋白来控制玉米螟害虫；与现有技术使用的农业防治方法、化学防治方法、物理防治方法和生物防治方法相比，本发明对植物进行全生育期、全植株的保护以防治玉米螟害虫的侵害，且无污染、无残留，效果稳定、彻底，简单、方便、经济。
最后所应说明的是，以上实施例仅用以说明本发明的技术方案而非限制，尽管参照较佳实施例对本发明进行了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的精神和范围。
序列表
<110> 北京大北农生物技术有限公司
<120> 杀虫蛋白的用途
<130> DBNBC161
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<170> SIPOSequenceListing 1.0
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<213> 人工序列-ACe1_3氨基酸序列(Artificial Sequence)
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Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Thr Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 2
<211> 295
<212> PRT
<213> 人工序列-ACe1_4氨基酸序列(Artificial Sequence)
<400> 2
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Ile Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 3
<211> 295
<212> PRT
<213> 人工序列-ACe1_5氨基酸序列(Artificial Sequence)
<400> 3
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Ser Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Tyr Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Ile Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 4
<211> 295
<212> PRT
<213> 人工序列-ACe1_6氨基酸序列(Artificial Sequence)
<400> 4
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Val Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Ile Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 5
<211> 295
<212> PRT
<213> 人工序列-ACe1_8氨基酸序列(Artificial Sequence)
<400> 5
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Val Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Tyr Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Ile Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 6
<211> 295
<212> PRT
<213> 人工序列-ACe1_9氨基酸序列(Artificial Sequence)
<400> 6
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Val Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Ser Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Tyr Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Ile Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 7
<211> 295
<212> PRT
<213> 人工序列-ACe1_10氨基酸序列(Artificial Sequence)
<400> 7
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp His Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Ile Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Glu Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 8
<211> 295
<212> PRT
<213> 人工序列-ACe1_11氨基酸序列(Artificial Sequence)
<400> 8
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Met
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Ser Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Thr Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 9
<211> 295
<212> PRT
<213> 人工序列-ACe1_12氨基酸序列(Artificial Sequence)
<400> 9
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asp Glu Ala Gly Thr Phe Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Val Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Ser Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asp Ser Ser Thr Asp Thr Thr Thr
145 150 155 160
Lys Thr Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Glu Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 10
<211> 295
<212> PRT
<213> 人工序列-ACe1_13氨基酸序列(Artificial Sequence)
<400> 10
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Val Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Ser Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Ile Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 11
<211> 295
<212> PRT
<213> 人工序列-ACe1_14氨基酸序列(Artificial Sequence)
<400> 11
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Met
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Ser Asp Gln Glu Leu Leu Thr Pro Glu Phe Thr Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Thr Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 12
<211> 295
<212> PRT
<213> 人工序列-ACe1_15氨基酸序列(Artificial Sequence)
<400> 12
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Ser Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asp Ser Ser Thr Asp Thr Thr Thr
145 150 155 160
Lys Thr Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 13
<211> 295
<212> PRT
<213> 人工序列-ACe1_16氨基酸序列(Artificial Sequence)
<400> 13
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Val Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Phe Ile Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser His Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Arg Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Ile Asp Gln Glu Met Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Gly Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Val Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Thr Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 14
<211> 295
<212> PRT
<213> 人工序列-ACe1_17氨基酸序列(Artificial Sequence)
<400> 14
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asp Glu Ala Gly Thr Phe Asn Glu Ala Trp Asn Ala Asn Phe Lys Pro
20 25 30
Ser Asp Glu Pro Gln Phe Ser Asn Ser Pro Thr Gln Gly Ile Val Ile
35 40 45
Leu Ala Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Arg Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Thr Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Arg Val
195 200 205
Ser Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asp
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Ser Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 15
<211> 295
<212> PRT
<213> 人工序列-ACe1_18氨基酸序列(Artificial Sequence)
<400> 15
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Val Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Phe Ile Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser His Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Arg Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Ile Asp Gln Glu Met Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Gly Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asn Ser Ser Thr Asp Thr Lys Thr
145 150 155 160
Lys Gln Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Val Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asp Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Thr Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 16
<211> 295
<212> PRT
<213> 人工序列-ACe1_19氨基酸序列(Artificial Sequence)
<400> 16
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Ala Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Val Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asp Ser Ser Thr Asp Thr Thr Thr
145 150 155 160
Lys Thr Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 17
<211> 295
<212> PRT
<213> 人工序列-ACe1_20氨基酸序列(Artificial Sequence)
<400> 17
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Val Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Leu Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asp Ser Ser Thr Asp Thr Thr Thr
145 150 155 160
Lys Thr Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Thr Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 18
<211> 295
<212> PRT
<213> 人工序列-ACe1_21氨基酸序列(Artificial Sequence)
<400> 18
Met Ser Ser Thr Asp Val Gln Glu Arg Leu Arg Asp Leu Ala Arg Glu
1 5 10 15
Asn Glu Ala Gly Thr Leu Asn Glu Ala Trp Asn Thr Asn Phe Lys Pro
20 25 30
Ser Asp Glu Gln Gln Phe Ser Tyr Ser Pro Thr Glu Gly Ile Val Phe
35 40 45
Leu Thr Pro Pro Lys Asn Val Ile Gly Glu Arg Arg Ile Ser Gln Tyr
50 55 60
Lys Val Asn Asn Ala Trp Ala Thr Leu Glu Gly Ser Pro Thr Glu Val
65 70 75 80
Ser Gly Thr Pro Leu Tyr Val Gly Lys Asn Val Leu Asp Asn Ser Lys
85 90 95
Gly Thr Met Asp Gln Glu Leu Leu Thr Pro Glu Phe Asn Tyr Thr Tyr
100 105 110
Thr Glu Ser Thr Ser Asn Thr Thr Thr His Gly Leu Lys Val Gly Val
115 120 125
Lys Thr Thr Ala Thr Met Lys Phe Pro Ile Ala Gln Gly Ser Met Glu
130 135 140
Ala Ser Thr Glu Tyr Asn Phe Gln Asp Ser Ser Thr Asp Thr Thr Thr
145 150 155 160
Lys Thr Val Ser Tyr Lys Ser Pro Ser Gln Lys Ile Lys Val Pro Ala
165 170 175
Gly Lys Thr Phe Arg Val Leu Ala Tyr Leu Asn Thr Gly Ser Ile Ser
180 185 190
Gly Glu Ala Asn Leu Tyr Ala Asn Val Gly Gly Ile Ala Trp Gly Val
195 200 205
Leu Pro Gly Tyr Pro Asn Gly Gly Gly Val Asn Ile Gly Ala Val Leu
210 215 220
Ser Lys Cys Gln Gln Lys Gly Trp Gly Asp Phe Arg Asn Phe Gln Pro
225 230 235 240
Ser Gly Arg Asp Val Ile Val Lys Gly Gln Gly Thr Phe Lys Ser Asn
245 250 255
Tyr Gly Thr Asp Phe Ile Leu Lys Ile Glu Asp Ile Thr Asp Ser Lys
260 265 270
Leu Arg Asn Asn Asn Gly Ser Gly Thr Val Val Gln Glu Ile Lys Val
275 280 285
Pro Leu Ile Arg Thr Glu Ile
290 295
<210> 19
<211> 888
<212> DNA
<213> 人工序列-ACe1_3核苷酸序列(Artificial Sequence)
<400> 19
atgagttcaa cagatgtaca agaaaggcta cgcgacctcg cgcgtgaaaa cgaggccggg 60
accttgaatg aggcttggaa taccaatttc aaaccgtccg atgaacagca gtttagctac 120
agcccgactg agggtatcgt gtttctgaca cctccgaaaa acgtgatcgg tgagcgccgt 180
attagccaat ataaagtcaa caacgcgtgg gcaacgctgg aaggttcccc gaccgaggtt 240
tcaggtacgc cattgtacgc gggtaagaac gtcctggata acagcaaggg tacgatggat 300
caagagctgc tgaccccgga atttaactac acctataccg aatcgacttc taataccaca 360
acccacggcc tgaaactggg tgttaagacc actgccacca tgaagtttcc gattgcacag 420
ggtagcatgg aggcgagcac ggagtataac ttccagaatt cctctacgga caccaagacc 480
aagcaggtta gctacaagag cccgtcgcaa aaaatcaagg tgccggcagg taaaacgttc 540
cgtgttttgg cgtatctgaa caccggtagc attagcggcg aggcgaatct gtacgcgaac 600
gtaggtggca ttgcgtgggg cgtcttaccg ggttatccga atggcggtgg cgtgaacatt 660
ggtgctgtgt tgaccaagtg ccagcaaaaa ggctggggtg atttccgcaa cttccaaccg 720
tctggccgtg acgtgattgt taaaggccaa ggcaccttca cctccaatta cggcactgac 780
tttatcctga agattgaaga tatcaccgac agcaaactgc gtaataacaa cggcagcggc 840
accgttgttc aggagatcaa agtgccactt atcagaaccg aaatctaa 888
<210> 20
<211> 888
<212> DNA
<213> 人工序列-ACe1_4核苷酸序列(Artificial Sequence)
<400> 20
atgagttcaa cagatgtaca agaaaggcta cgcgacctgg cgagagaaaa tgaagcgggt 60
acgttaaacg aggcttggaa taccaatttc aaaccgagcg atgaacaaca atttagctac 120
agcccgaccg agggcatcgt gtttctgacc ccgcctaaaa acgtcatcgg cgaacgtcgt 180
atcagccagt ataaagtcaa caacgcgtgg gcaaccctcg aaggcagccc aacggaggtc 240
agcggcaccc cgttgtacgc tggcaagaac gtgctggata atagcaaagg caccatggat 300
caagagttgc tgaccccgga atttaattac acctatactg agagcacctc aaacaccact 360
acgcacggcc tgaaactggg tgttaaaacc accgcaacca tgaaatttcc gattgctcag 420
ggttccatgg aagcgagtac cgagtataac ttccaaaact ccagcacgga caccaagacc 480
aagcaggttt cctacaaaag cccgtcgcaa aaaatcaagg tgccggcagg caagaccttc 540
cgcgtattgg cgtatctgaa taccggttcc atttctggcg aggcgaacct gtacgcgaat 600
gtgggtggta tcgcctgggg tgtgctgccg ggctatccga atggtggtgg cattaacatc 660
ggtgccgttt tgacgaagtg ccagcagaag ggttggggtg attttcgtaa cttccagccg 720
tctggtcgtg acgttattgt gaagggtcag ggtactttca agtcgaacta cggcacggac 780
ttcatcctga agatcgagga tattaccgac agcaaactgc gtaacaacaa cggctctggt 840
acagttgttc aggagatcaa agtgccactt attcgcaccg aaatttaa 888
<210> 21
<211> 888
<212> DNA
<213> 人工序列-ACe1_5核苷酸序列(Artificial Sequence)
<400> 21
atgagttcaa cagatgtaca agaaaggcta cgcgacctgg cgcgtgaaaa cgaggcgggc 60
acgttgaacg aggcgtggaa taccaatttt aaaccgtcgg atgaacagca gtttagctac 120
agcccgacag agggtatcgt gtttctgacc ccgcctaaaa acgtgatcgg tgaacgccgt 180
atcagccagt ataaggtgtc taacgcttgg gcaaccctcg aaggcagccc gaccgaggtc 240
tccggcactc cattgtacgc gggaaagaac gtgttggata attctaaggg caccatggat 300
caagagctgc tgactccaga atttaactac acctacaccg agagcaccag caacactacc 360
acccacggcc tgaagctggg tgtgaagacg accgcaacca tgaaattccc gattgcgcag 420
ggttccatgg aagcgagcac ggaatataac ttccaaaaca gcagcaccga taccaagacg 480
aagcaggtta gctacaaatc cccgtcgcaa aaaatcaaag ttccggcagg taagacctat 540
cgtgttcttg cctatctgaa tacgggtagt atcagcggtg aggcgaacct gtatgctaat 600
gttggtggca tcgcctgggg tgttctgccg ggttacccga atggtggtgg tattaatatt 660
ggcgctgtgt tgaccaagtg ccagcaaaaa ggttggggcg acttccgcaa cttccaaccg 720
tctggccgtg acgttattgt gaagggtcag ggtacgttta aatccaacta cggcaccgac 780
ttcatcctga aaattgaaga tattaccgac agcaaattac gtaataacaa cggctccggc 840
actgtagttc aggagatcaa ggtcccgctg attcgtaccg agatctaa 888
<210> 22
<211> 888
<212> DNA
<213> 人工序列-ACe1_6核苷酸序列(Artificial Sequence)
<400> 22
atgagttcaa cagatgtaca agaaaggcta cgcgacctgg cacgtgaaaa cgaagctggc 60
accctcaacg ttgcgtggaa cacgaacttc aaaccgagcg atgaacagca atttagctac 120
agcccgaccg agggcattgt ttttctgacg ccgccaaaaa acgtgattgg tgaacgtcgt 180
attagccagt ataaagtgaa taatgcttgg gcaaccttgg aaggtagtcc aactgaggtt 240
agcggcaccc cgctgtacgc cggtaagaac gttttagata atagcaaggg cacaatggat 300
caagagttgc tgaccccgga atttaactat acctacaccg agagcaccag caacaccacg 360
acccacggcc tgaaacttgg tgttaaaacc accgcgacca tgaaattccc gattgcgcag 420
ggctcgatgg aagcgagcac cgagtataac ttccaaaact ccagcactga caccaagact 480
aagcaggtgt cttacaagag cccgtcgcaa aaaatcaagg tgcctgcagg caagaccttt 540
cgtgttttgg cgtatctgaa taccggttcc atctctggcg aggccaatct gtatgcgaac 600
gtgggtggca tcgcgtgggg tgtgctgccg ggttacccga atggcggtgg tatcaacatt 660
ggcgctgtgt tgaccaagtg ccaacagaag ggatggggtg atttccgcaa ctttcaaccg 720
tccggccgtg acgtcatcgt gaaaggtcag ggtacgttca aatccaatta cggtacggac 780
ttcatcctga agatcgaaga tattaccgac agcaaactga gaaataacaa cggttctggt 840
actgtagtcc aggagatcaa agttccgctg attcgcaccg agatctaa 888
<210> 23
<211> 888
<212> DNA
<213> 人工序列-ACe1_8核苷酸序列(Artificial Sequence)
<400> 23
atgagttcaa cagatgtaca agaaaggcta cgcgacctgg cgcgtgaaaa tgaagcgggc 60
acgctgaatg tcgcttggaa cacgaacttc aagccgagcg atgaacagca attttcctac 120
agcccaactg aaggtattgt gtttctgacc ccaccgaaaa acgttatcgg cgagcgccgt 180
atttcccaat ataaggttaa taacgcgtgg gcaacattgg aaggttcccc gaccgaggtg 240
agtggcaccc cgctgtacgc tggcaagaac gtgttggaca acagcaaagg caccatggat 300
caagagttgc tgaccccgga attcaattac acctacaccg agagcaccag caacactacg 360
acccacggtc tgaagctggg tgtgaaaacc accgcaacca tgaaatttcc gattgcccag 420
ggtagcatgg aagcgagcac ggagtataat ttccaaaact cgtcaaccga taccaagacc 480
aagcaggttt cctacaagag cccgtctcaa aaaattaaag taccggcagg caagacctat 540
cgtgttttag cgtatctgaa caccggtagc atttctggcg aggcgaacct gtacgccaat 600
gtgggtggca tcgcgtgggg agtgctcccg ggttatccga atggtggcgg gatcaacatt 660
ggcgctgtgt tgacgaagtg ccagcagaaa ggttggggtg atttccgtaa tttccagccg 720
agtggtcgtg acgttatcgt taaaggccaa ggtacgttta aaagcaatta cggtactgac 780
ttcatcctga aaatcgaaga tatcaccgac agcaaactaa gaaacaacaa cggttctggc 840
accgttgtcc aggagatcaa ggtcccgctt atccgcaccg agatttaa 888
<210> 24
<211> 888
<212> DNA
<213> 人工序列-ACe1_9核苷酸序列(Artificial Sequence)
<400> 24
atgagttcaa cagatgtaca agaaaggcta cgcgacttag cgcgtgaaaa tgaagcgggt 60
acgttgaacg tggcctggaa taccaacttc aaaccgtccg acgaacagca gttttcttac 120
agcccgacgg agggtattgt gtttctgact ccaccgaaaa acgtgatcgg tgagcgccgt 180
attagccagt ataaggtaaa taacgcttgg gcaacccttg agggctcgcc taccgaggtt 240
tctggtacgc cgctgtatgc gggcaaaaac gtgctggata acagcaaagg cacttcagat 300
caggagctgc tgaccccgga attcaactac acgtacaccg aatctacctc caacaccacc 360
acgcacggcc tgaaactggg tgttaagact accgcgacca tgaaattccc gattgcacag 420
ggttccatgg aagcgagcac cgagtataac ttccaaaact caagcactga caccaagacc 480
aagcaggtta gctacaagag cccatcgcaa aaaatcaagg ttccggcagg caaaacctac 540
cgtgtcttag cctatctgaa taccggttcc attagcggcg aggcgaatct gtacgcgaac 600
gttggtggca tcgcttgggg ggtgttgccg ggttatccga atggaggcgg tatcaacatt 660
ggcgctgtgt tgacgaagtg ccagcaaaaa ggttggggcg attttagaaa ctttcagccg 720
tccggtcgtg atgttattgt gaagggccaa ggcaccttta agagcaatta cggcaccgac 780
ttcatcctga agatcgaaga tatcaccgac agcaaattgc gcaacaacaa tggtagcggt 840
acagtcgttc aagagatcaa agtgccgctg attcgtaccg aaatctaa 888
<210> 25
<211> 888
<212> DNA
<213> 人工序列-ACe1_10核苷酸序列(Artificial Sequence)
<400> 25
atgagttcaa cagatgtaca agaaaggcta cgcgacttgg cgcgtgaaaa cgaggcgggc 60
accttgaacg aggcctggca caccaatttc aaaccgagcg atgaacagca gtttagctac 120
agcccgaccg agggtattgt ctttctgacc ccgccaaaga atgtcattgg cgaacgtcgt 180
atctcgcaat ataaagttaa taacgcgtgg gcaaccttgg aaggcagtcc gaccgaagtg 240
agcggcactc cgctgtacgc aggcaaaaac gtgctggaca acagcaaagg caccatggat 300
caggagctgc tgactccgga attcaactac acctataccg agtctacgtc aaataccatt 360
acccatggtc tgaagctcgg cgtgaagacg acggcaacca tgaaatttcc gattgctcaa 420
ggcagcatgg aagcgagcac cgagtataac ttccaaaact cttccactga caccaaaacc 480
aagcaggttt cctacaaaag ccctagccaa aagattaagg tgccggctgg caaaacattt 540
cgtgttttgg cgtatctgaa taccggttcc atcagcggtg aagcgaatct gtacgcgaac 600
gtgggtggca tcgcctgggg tgtgcttccg ggttacccga atggtggcgg agtgaacatc 660
ggcgctgttt tgacgaagtg ccagcagaag ggttggggtg atttccgcaa tttccaaccg 720
tccggtagag atgttatcgt aaaaggtcag ggtacatttg agtcgaacta tggtacggac 780
ttcatcctga agatcgaaga tatcaccgac agcaaactgc gcaacaacaa cggttctggc 840
accgttgttc aagagattaa ggtgccgctg attcgtaccg agatctaa 888
<210> 26
<211> 888
<212> DNA
<213> 人工序列-ACe1_11核苷酸序列(Artificial Sequence)
<400> 26
atgagttcaa cagatgtaca agaaaggcta cgcgatctgg cgcgtgaaaa tgaggccggc 60
accttgaacg aagcgtggaa taccaatttc aaaccgtccg acgaacagca atttagctac 120
agcccgaccg agggcatcgt ttttctgact cctccgaaaa acgtcattgg cgaacgtcgt 180
atttctcagt ataaggtgaa caacgcatgg gcaaccttgg aaggttcccc gacggagatg 240
agcggcacgc cattgtacgc cggtaagaac gtgctggata acagcaaggg cacaagcgac 300
caagagctgc tcaccccgga atttaactac acgtataccg agagcacctc gaacacgacc 360
acccacggcc tgaagctggg tgtgaaaacg accgctacca tgaaattccc gattgcgcag 420
ggtagcatgg aagcgtcaac tgagtacaat ttccaaaaca gcagcaccga taccaagact 480
aaacaggtta gctacaagtc cccgtcgcag aaaatcaaag ttccggcagg taaaaccttt 540
cgtgttctgg cgtatctgaa caccggtagt atttccggtg aggcgaacct gtacgctaac 600
gtgggtggca tcgcgtgggg tgtattgcca ggttatccga atggtggcgg agttaatatt 660
ggtgctgtgc ttaccaagtg ccaacaaaag ggttggggcg atttccgcaa cttccagccg 720
tccggccgtg acgtgatcgt taaaggccag ggcaccttta ccagtaatta tggtactgac 780
ttcatcctga agatcgaaga tatcaccgac agcaaactgc gcaataacaa cggctctggg 840
acggttgtcc aggagatcaa ggtgccgtta attagaaccg agatttaa 888
<210> 27
<211> 888
<212> DNA
<213> 人工序列-ACe1_12核苷酸序列(Artificial Sequence)
<400> 27
atgagttcaa cagatgtaca agaaaggcta cgtgatttgg cgcgtgaaga tgaagcgggc 60
accttcaatg aagcgtggaa tactaatttt aagccgtctg acgagcagca atttagctat 120
tccccgactg agggcattgt ctttctgacc ccaccgaaga acgtgattgg tgaacgtaga 180
atcagccagt ataaagtcaa caatgcgtgg gcaaccctgg aaggtagccc gaccgaagtt 240
agcggcaccc cgctatatgt tggcaaaaac gtacttgata atagcaaagg cacatccgac 300
caggagttgc tgactccgga attcaactat acctacaccg agagcacctc taacaccaca 360
acccacggcc tgaagctggg tgtgaagacc accgctacga tgaaatttcc gatagcccaa 420
ggtagcatgg aagcgagcac cgagtacaat ttccaagaca gctcgacgga taccacgacg 480
aagacggtta gctacaaatc tccgtcccag aaaatcaaag ttccggcagg taagaccttc 540
cgcgtgctcg catacctgaa taccggtagc atcagcggcg aggctaacct gtatgcgaac 600
gttggcggca tcgcgtgggg tgtgctgccg ggttacccga atggtggcgg tgttaacatt 660
ggtgccgtgt tgaccaagtg ccagcaaaag ggttggggcg agttccgcaa cttccagcct 720
tctggccgtg atgttattgt gaaaggtcag ggtacgttca agtccaacta cggcaccgac 780
tttattctga agatcgaaga catcaccgac tccaaactgc gtaacaacaa cggcagtggc 840
accgtcgtgc aagagatcaa agtgccattg attcgcaccg agatctaa 888
<210> 28
<211> 888
<212> DNA
<213> 人工序列-ACe1_13核苷酸序列(Artificial Sequence)
<400> 28
atgagttcaa cagatgtaca agaaaggcta cgcgatctgg cgcgtgaaaa cgaagctggc 60
accttgaatg tggcgtggaa cacgaacttc aagccgagcg atgagcagca atttagctat 120
tctccgaccg aaggtatcgt gttcttgacc ccgcctaaaa acgtcatcgg tgagcgtaga 180
atcagccagt ataaggttaa taatgcttgg gcaaccctgg aaggttcccc aaccgaagtt 240
agtggtacgc cactttacgc gggtaaaaac gtgctggata actcaaaagg cacgtccgac 300
caggagctgt taaccccgga atttaattac acctacaccg agagcacttc caatacgacc 360
acgcacggcc tgaaactggg tgttaaaacc accgccacca tgaagtttcc gattgctcag 420
ggctctatgg aggccagcac cgagtataat ttccaaaaca gcagcaccga cactaaaacg 480
aagcaagtta gctacaagtc cccgtcgcaa aaaatcaaag ttccggcggg taagactttt 540
cgtgtattgg cgtatctgaa caccggtagc atttctggcg aggcgaatct gtacgcgaac 600
gtcggcggca ttgcatgggg tgttctaccg ggttatccga atggcggtgg tatcaatatc 660
ggtgcagtcc tgacaaagtg ccaacagaag ggctggggtg atttccgcaa cttccagccg 720
tccggccgtg acgtgattgt taagggtcag ggcaccttta aaagcaacta cggcaccgat 780
ttcatcttga aaattgaaga cattaccgac agcaagctgc gcaacaacaa cggctctggc 840
actgtggtgc aagaaatcaa ggtgccgctg atccgtaccg agatttaa 888
<210> 29
<211> 888
<212> DNA
<213> 人工序列-ACe1_14核苷酸序列(Artificial Sequence)
<400> 29
atgagttcaa cagatgtaca agaaaggcta agagacctgg cgcgtgaaaa cgaggcggga 60
accttgaatg aggcttggaa taccaatttc aaaccgtccg atgaacagca gtttagctac 120
agcccgaccg agggcatcgt gtttctgacc ccgcctaaaa acgtcatcgg cgaacgtcgt 180
atctcgcaat ataaagtgaa caacgcttgg gcaaccttag aaggttcccc gaccgagatg 240
agcggcaccc cgttgtacgc aggcaagaac gtgcttgata acagcaaagg cacgtccgac 300
caagagctgc tgacgccaga atttacgtac acgtacactg agagcaccag caacacgacc 360
acccacggcc tgaaactggg tgtgaagacc actgccacca tgaaatttcc gattgcccag 420
ggctctatgg aagcgagcac tgagtataac ttccaaaaca gcagcactga caccaagacc 480
aagcaggtta gctacaaatc cccatctcag aaaattaagg tgccggcagg taaaaccttc 540
cgcgtgctcg cgtatctgaa caccggttcg atctctggcg aggcgaatct gtacgcgaac 600
gttggtggta ttgcgtgggg tgtactgccg ggttatccga atggtggcgg tgttaatatt 660
ggtgctgtgt tgaccaagtg ccagcagaag ggctggggtg atttccgcaa ctttcaaccg 720
agtggccgtg atgttattgt taagggtcag ggtacattca cctctaatta tggtacggac 780
ttcatcttga aaatcgaaga tattaccgac agcaagctgc gcaataacaa cggcagcggc 840
accgttgttc aagaaatcaa ggtcccgctg attcgtaccg agatctaa 888
<210> 30
<211> 888
<212> DNA
<213> 人工序列-ACe1_15核苷酸序列(Artificial Sequence)
<400> 30
atgagttcaa cagatgtaca agaaaggcta cgcgacctag cgcgtgaaaa tgaagctggc 60
accctgaatg aagcgtggaa cacgaacttc aaaccgtccg acgaacaaca attttcctac 120
agccctaccg agggtatcgt gtttctgacc ccaccgaaga acgtgatcgg cgaacgccgt 180
attagccaat ataaggtgaa taacgcgtgg gcaaccttgg agggctcccc gaccgaggtt 240
tctggcacac cgttgtatgc tggtaaaaac gtgttggaca acagcaaggg caccagcgat 300
caggagctgc tgacgccgga atttaactac acctataccg agagcacctc aaataccacc 360
acccacggcc tgaagctggg tgtcaaaacc actgccacca tgaagtttcc gattgcgcag 420
ggcagcatgg aagctagcac cgagtacaat ttccaagata gcagcaccga tactacgacc 480
aaaaccgtta gctacaaatc tccgtcgcaa aagatcaaag tgccggcagg caagacgttt 540
cgtgttttag cgtatctgaa caccggttcc atttctggtg aggcgaacct gtacgccaat 600
gttggtggga tcgcgtgggg tgttctgccg ggttatccga atggtggcgg tgttaacatt 660
ggcgcagttt tgaccaagtg ccagcagaaa ggctggggtg atttccgcaa tttccagccg 720
tcgggccgtg acgtgatcgt taaaggtcag ggtacattca agagcaacta cggcacggac 780
ttcatcctga aaattgaaga tattaccgac agcaaactga gaaacaacaa cggtagtggc 840
acggtcgtgc aggagatcaa ggtcccgctc atccgtactg agatctaa 888
<210> 31
<211> 888
<212> DNA
<213> 人工序列-ACe1_16核苷酸序列(Artificial Sequence)
<400> 31
atgagttcaa cagatgtaca agaaaggcta cgcgacctgg cccgtgaaaa cgaggcgggc 60
acgctgaacg tcgcgtggaa tacgaacttc aaaccgtcag atgaacagca atttagctac 120
agcccgacgg agggttttat ctttctgacc ccaccgaaaa acgtgatcgg cgagcgccgt 180
atttctcact ataaagtgaa caacgcatgg gcgaccttgg aaggtagccc gaccgaggtt 240
agcggcacgc cgctgtacgc gggtcgtaac gtgctggata attccaaagg caccattgat 300
caagaaatgt tgacccctga atttaactac acctataccg aggggacctc taacaccacc 360
acgcatggtc tgaaacttgg tgtcaagacc acagcgacca tgaaattccc gattgcacag 420
ggtagcatgg aagcgagcac cgagtataac ttccaaaaca gtagcactga caccaagacg 480
aagcaggtta gctacaagag cccgtcccag aaaatcaaag ttccggcagg taaaaccttc 540
cgcgtgttgg cctatctgaa taccggttcc attagcggcg aagcgaatct ctacgctaat 600
gtgggcggtg tagcctgggg tgtgctgcca ggttatccga atggcggcgg cgtcaacatt 660
ggtgctgtgt tgaccaagtg ccagcaaaag ggctggggcg atttccgtaa cttccagccg 720
tctggtagag acgttattgt taagggtcag ggcacgttta ccagcaatta cggcactgac 780
ttcatcctga agatcgaaga tatcaccgac tccaaactgc gtaataacaa cggttcgggc 840
acagttgttc aagagatcaa ggtgccgtta attcgtaccg agatctaa 888
<210> 32
<211> 888
<212> DNA
<213> 人工序列-ACe1_17核苷酸序列(Artificial Sequence)
<400> 32
atgagttcaa cagatgtaca agaaaggcta agagatctgg cgcgtgaaga cgaagctggc 60
acgtttaacg aagcgtggaa tgcaaacttt aaaccgtccg acgagccgca gtttagcaac 120
agtccgacac agggcatcgt gatcttggcc cctccgaaaa acgtgatcgg tgaacgtcgt 180
attagccaat ataaagtaaa taacgcgtgg gcaaccctgg aaggttcccc aaccgaggtt 240
agcggcaccc cgttgtacgc gggtcgcaac gtgcttgata attccaaagg caccatggat 300
caggagctgc tgaccccgga attcacgtac acctataccg agagcacctc taacaccacc 360
actcacggcc tgaagctggg tgtgaaaacg actgccacca tgaaatttcc gattgcccaa 420
ggttctatgg aagcgagcac tgagtacaat ttccaaaact cgagcaccga caccaagacg 480
aagcaggtta gctataaatc cccaagccag aaaatcaagg tgccggctgg gaaaaccttt 540
cgtgtgttag cgtatctgaa taccggttcg atttccggcg aggcgaacct gtacgctaac 600
gttggtggta ttgcgtggcg tgttagcccg ggttatccga atggtggtgg tgtcaatatt 660
ggcgcagttc tcacgaagtg ccagcaaaag ggctggggtg atttccgcaa cttccaaccg 720
agcggccgtg atgttatcgt gaagggccag ggtacgttca agtccgacta cggcactgac 780
ttcatcttga agattgaaga tatcaccgac agcaagtctc gtaacaacaa cggttctggc 840
accgtcgttc aggagatcaa agttccgctg attcgcaccg agatctaa 888
<210> 33
<211> 888
<212> DNA
<213> 人工序列-ACe1_18核苷酸序列(Artificial Sequence)
<400> 33
atgagttcaa cagatgtaca agaaaggcta cgcgatctgg cgcgtgaaaa cgaagctggc 60
accttaaacg tggcctggaa tacgaacttc aaaccttcag acgagcaaca atttagctac 120
agcccaacgg aaggcttcat ctttctgacc ccgccaaaaa acgtcattgg tgagcgtcgt 180
atttcccatt ataaggttaa caacgcttgg gcgactttgg aaggttcgcc gaccgaggtg 240
agcggtactc cgttgtacgc aggccgcaat gtgctggata attccaaagg tacgattgac 300
caggagatgc tgactccgga atttaactac acctatactg agggcacctc taataccacc 360
acgcacggtc tgaagctggg tgtaaaaacc accgcaacga tgaaattccc gatcgcgcag 420
gggtctatgg aagcgagcac cgagtataat tttcaaaatt ccagcaccga caccaagacc 480
aagcaggttt cctacaagtc gccgagccag aaaatcaaag ttccggcggg taaaaccttt 540
cgtgtcttgg cgtatctgaa taccggtagc atttctggag aggcgaacct gtacgccaac 600
gttggtggcg ttgcatgggg tgtgcttccg ggttatccgg atggcggtgg cgttaatatc 660
ggcgctgtgt tgaccaagtg ccagcaaaag ggttggggcg atttccgcaa cttccagccg 720
tccggtagag acgtgatcgt gaaaggtcag ggcacattca ccagcaacta cggcacggac 780
ttcattctca agattgaaga tatcaccgac agcaaactgc gtaataacaa cggtagcggc 840
accgttgttc aagaaatcaa ggtgccgctg attcgtaccg agatctaa 888
<210> 34
<211> 888
<212> DNA
<213> 人工序列-ACe1_19核苷酸序列(Artificial Sequence)
<400> 34
atgagttcaa cagatgtaca agaaaggcta cgcgacctgg cgcgtgaaaa cgaggccggt 60
actctgaacg aggcttggaa taccaatttc aaaccgtctg atgagcaaca atttagctac 120
agcccgactg agggtattgt gtttttgact ccaccaaaaa acgtgatcgg tgaacgtcgt 180
atttcccagt ataaggtaaa caacgcgtgg gcaaccctgg aaggttcacc tacagaggtt 240
agcggcacgc cgctgtacgc cggtaagaac gttctggaca acagcaaggg tacgatggat 300
caagagttgc tgaccccgga atttaactac acctataccg agtccaccag caacactacg 360
acccacggcc tgaaggtggg cgtcaaaacc accgcaacca tgaaatttcc gattgcgcag 420
ggttctatgg aagcgagcac cgaatataac ttccaagatt ccagcaccga caccactacg 480
aagaccgttt cctacaagag cccgtcgcaa aaaatcaagg tgccggcagg caaaaccttc 540
agagtcctcg cgtatttgaa taccggttcg atcagcggtg aagcgaactt atatgcgaat 600
gttgggggca ttgcttgggg cgtgctgccg ggttacccga atggtggcgg tgtgaatatt 660
ggtgctgtgt tgaccaagtg ccagcagaaa ggctggggtg atttccgcaa cttccagccg 720
agcggccgtg acgttattgt taaaggtcag ggcacgttta agtcgaatta cggcactgac 780
ttcatcctga agatcgaaga tattaccgac agcaaactgc gtaataacaa cggcagtggc 840
acagttgttc aggagatcaa agtcccgctt atccgcaccg agatctaa 888
<210> 35
<211> 888
<212> DNA
<213> 人工序列-ACe1_20核苷酸序列(Artificial Sequence)
<400> 35
atgagttcaa cagatgtaca agaaaggcta cgcgacttgg cgcgtgaaaa cgaagctggc 60
acccttaatg aagcctggaa tacgaacttc aaaccgtctg atgagcagca gtttagctac 120
agcccgaccg aaggcattgt ttttctgacc ccaccgaaaa acgtaatcgg cgaaagacgt 180
atttcgcaat ataaagtgaa caacgcatgg gcaaccttgg aaggttcccc gaccgaggtg 240
tctggtactc cgctgtacgt cggcaaaaat gtgctggata atagcaaagg tacaatggat 300
caggagctgc tgaccccgga atttaactac acctataccg agtccaccag caataccact 360
acgcacggct taaagctggg tgtgaaaacc acggctacga tgaaatttcc gattgcgcag 420
ggctccatgg aagcgtccac cgagtacaat ttccaagaca gcagcaccga caccactacg 480
aaaaccgtta gctacaagtc gccgagtcaa aaaattaagg tgccggcggg taaaacgttc 540
cgcgttctcg cgtatctgaa caccggaagc atttccggcg aggccaacct gtacgcgaac 600
gttggtggca tcgcttgggg tgtgctgccg ggttatccga atggtggtgg cgttaacatt 660
ggtgcagttt tgaccaagtg ccagcaaaag ggctggggcg atttccgcaa cttccagcca 720
agcggccgtg acgtcatcgt gaaaggtcag ggcaccttta agtctaacta tggtacggac 780
ttcatcttga agatcgagga catcaccgat agcaagctgc gtaataacaa cggttcaggt 840
actgtcgttc aagagatcaa ggtgcctctg attcgtaccg agatctaa 888
<210> 36
<211> 888
<212> DNA
<213> 人工序列-ACe1_21核苷酸序列(Artificial Sequence)
<400> 36
atgagttcaa cagatgtaca agaaaggcta cgcgacctgg cgcgtgaaaa cgaggccggt 60
acgttgaatg aagcgtggaa tacgaacttt aaaccgtctg atgaacagca atttagctac 120
agcccgaccg agggcattgt gtttctgact ccgcctaaaa acgtcatcgg cgaacgtcgt 180
attagccaat ataaagtcaa taacgcgtgg gcaaccttgg aaggctctcc gaccgaagtt 240
tctggcaccc cgttgtacgt tggtaaaaac gtgctggaca atagcaaggg tacgatggat 300
caggagctgc tgaccccaga atttaactac acctataccg agagcaccag caacaccacc 360
acccacggcc tgaaggtggg tgtgaagacc accgcaacca tgaaattccc gattgcgcag 420
ggttccatgg aagcgagcac ggagtataac ttccaagata gcagcactga caccactaca 480
aaaaccgtat cctacaagag cccgtcgcaa aagatcaagg ttccggcagg caagaccttc 540
cgcgtgcttg cctacctgaa caccggtagc attagtggcg aggctaacct gtatgcgaac 600
gttggtggta tcgcgtgggg ggtgctcccg ggttatccga atggtggtgg cgttaatatt 660
ggcgctgtgt tgtccaagtg ccagcaaaaa ggttggggtg atttccgcaa tttccagccg 720
tcgggccgtg acgttatcgt taaaggtcag ggtacgttta agtctaatta cggcactgac 780
ttcatcctga agatcgaaga tattaccgac tccaaactgc gtaacaacaa cggttcaggc 840
acggtggttc aggagatcaa agtcccactg attcgtaccg agatctaa 888
<210> 37
<211> 818
<212> PRT
<213> 人工序列-Cry1Ab氨基酸序列(Artificial Sequence)
<400> 37
Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu
1 5 10 15
Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly
20 25 30
Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser
35 40 45
Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile
50 55 60
Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile
65 70 75 80
Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala
85 90 95
Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu
100 105 110
Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu
115 120 125
Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala
130 135 140
Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val
145 150 155 160
Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser
165 170 175
Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg
180 185 190
Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val
195 200 205
Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg
210 215 220
Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val
225 230 235 240
Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro
245 250 255
Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val
260 265 270
Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu
275 280 285
Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr
290 295 300
Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln
305 310 315 320
Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro
325 330 335
Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala
340 345 350
Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg
355 360 365
Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp
370 375 380
Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val
385 390 395 400
Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln
405 410 415
Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His
420 425 430
Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile
435 440 445
Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Glu Phe Asn Asn
450 455 460
Ile Ile Pro Ser Ser Gln Ile Thr Gln Ile Pro Leu Thr Lys Ser Thr
465 470 475 480
Asn Leu Gly Ser Gly Thr Ser Val Val Lys Gly Pro Gly Phe Thr Gly
485 490 495
Gly Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln Ile Ser Thr Leu Arg
500 505 510
Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr Arg Val Arg Ile Arg
515 520 525
Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser Ile Asp Gly Arg
530 535 540
Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr Met Ser Ser Gly Ser Asn
545 550 555 560
Leu Gln Ser Gly Ser Phe Arg Thr Val Gly Phe Thr Thr Pro Phe Asn
565 570 575
Phe Ser Asn Gly Ser Ser Val Phe Thr Leu Ser Ala His Val Phe Asn
580 585 590
Ser Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu Phe Val Pro Ala Glu
595 600 605
Val Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg Ala Gln Lys Ala Val
610 615 620
Asn Glu Leu Phe Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val
625 630 635 640
Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Glu Cys Leu Ser
645 650 655
Asp Glu Phe Cys Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys
660 665 670
His Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn
675 680 685
Phe Arg Gly Ile Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr
690 695 700
Asp Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val
705 710 715 720
Thr Leu Leu Gly Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln
725 730 735
Lys Ile Asp Glu Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg
740 745 750
Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr
755 760 765
Asn Ala Lys His Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp
770 775 780
Pro Leu Ser Ala Pro Ser Pro Ile Gly Lys Cys Ala His His Ser His
785 790 795 800
His Phe Ser Leu Asp Ile Asp Val Gly Cys Thr Asp Leu Asn Glu Asp
805 810 815
Phe Arg
<210> 38
<211> 634
<212> PRT
<213> 人工序列-Cry2Ab氨基酸序列(Artificial Sequence)
<400> 38
Met Asp Asn Ser Val Leu Asn Ser Gly Arg Thr Thr Ile Cys Asp Ala
1 5 10 15
Tyr Asn Val Ala Ala His Asp Pro Phe Ser Phe Gln His Lys Ser Leu
20 25 30
Asp Thr Val Gln Lys Glu Trp Thr Glu Trp Lys Lys Asn Asn His Ser
35 40 45
Leu Tyr Leu Asp Pro Ile Val Gly Thr Val Ala Ser Phe Leu Leu Lys
50 55 60
Lys Val Gly Ser Leu Val Gly Lys Arg Ile Leu Ser Glu Leu Arg Asn
65 70 75 80
Leu Ile Phe Pro Ser Gly Ser Thr Asn Leu Met Gln Asp Ile Leu Arg
85 90 95
Glu Thr Glu Lys Phe Leu Asn Gln Arg Leu Asn Thr Asp Thr Leu Ala
100 105 110
Arg Val Asn Ala Glu Leu Thr Gly Leu Gln Ala Asn Val Glu Glu Phe
115 120 125
Asn Arg Gln Val Asp Asn Phe Leu Asn Pro Asn Arg Asn Ala Val Pro
130 135 140
Leu Ser Ile Thr Ser Ser Val Asn Thr Met Gln Gln Leu Phe Leu Asn
145 150 155 160
Arg Leu Pro Gln Phe Gln Met Gln Gly Tyr Gln Leu Leu Leu Leu Pro
165 170 175
Leu Phe Ala Gln Ala Ala Asn Leu His Leu Ser Phe Ile Arg Asp Val
180 185 190
Ile Leu Asn Ala Asp Glu Trp Gly Ile Ser Ala Ala Thr Leu Arg Thr
195 200 205
Tyr Arg Asp Tyr Leu Lys Asn Tyr Thr Arg Asp Tyr Ser Asn Tyr Cys
210 215 220
Ile Asn Thr Tyr Gln Ser Ala Phe Lys Gly Leu Asn Thr Arg Leu His
225 230 235 240
Asp Met Leu Glu Phe Arg Thr Tyr Met Phe Leu Asn Val Phe Glu Tyr
245 250 255
Val Ser Ile Trp Ser Leu Phe Lys Tyr Gln Ser Leu Leu Val Ser Ser
260 265 270
Gly Ala Asn Leu Tyr Ala Ser Gly Ser Gly Pro Gln Gln Thr Gln Ser
275 280 285
Phe Thr Ser Gln Asp Trp Pro Phe Leu Tyr Ser Leu Phe Gln Val Asn
290 295 300
Ser Asn Tyr Val Leu Asn Gly Phe Ser Gly Ala Arg Leu Ser Asn Thr
305 310 315 320
Phe Pro Asn Ile Val Gly Leu Pro Gly Ser Thr Thr Thr His Ala Leu
325 330 335
Leu Ala Ala Arg Val Asn Tyr Ser Gly Gly Ile Ser Ser Gly Asp Ile
340 345 350
Gly Ala Ser Pro Phe Asn Gln Asn Phe Asn Cys Ser Thr Phe Leu Pro
355 360 365
Pro Leu Leu Thr Pro Phe Val Arg Ser Trp Leu Asp Ser Gly Ser Asp
370 375 380
Arg Glu Gly Val Ala Thr Val Thr Asn Trp Gln Thr Glu Ser Phe Glu
385 390 395 400
Thr Thr Leu Gly Leu Arg Ser Gly Ala Phe Thr Ala Arg Gly Asn Ser
405 410 415
Asn Tyr Phe Pro Asp Tyr Phe Ile Arg Asn Ile Ser Gly Val Pro Leu
420 425 430
Val Val Arg Asn Glu Asp Leu Arg Arg Pro Leu His Tyr Asn Glu Ile
435 440 445
Arg Asn Ile Ala Ser Pro Ser Gly Thr Pro Gly Gly Ala Arg Ala Tyr
450 455 460
Met Val Ser Val His Asn Arg Lys Asn Asn Ile His Ala Val His Glu
465 470 475 480
Asn Gly Ser Met Ile His Leu Ala Pro Asn Asp Tyr Thr Gly Phe Thr
485 490 495
Ile Ser Pro Ile His Ala Thr Gln Val Asn Asn Gln Thr Arg Thr Phe
500 505 510
Ile Ser Glu Lys Phe Gly Asn Gln Gly Asp Ser Leu Arg Phe Glu Gln
515 520 525
Asn Asn Thr Thr Ala Arg Tyr Thr Leu Arg Gly Asn Gly Asn Ser Tyr
530 535 540
Asn Leu Tyr Leu Arg Val Ser Ser Ile Gly Asn Ser Thr Ile Arg Val
545 550 555 560
Thr Ile Asn Gly Arg Val Tyr Thr Ala Thr Asn Val Asn Thr Thr Thr
565 570 575
Asn Asn Asp Gly Val Asn Asp Asn Gly Ala Arg Phe Ser Asp Ile Asn
580 585 590
Ile Gly Asn Val Val Ala Ser Ser Asn Ser Asp Val Pro Leu Asp Ile
595 600 605
Asn Val Thr Leu Asn Ser Gly Thr Gln Phe Asp Leu Met Asn Ile Met
610 615 620
Leu Val Pro Thr Asn Ile Ser Pro Leu Tyr
625 630
<210> 39
<211> 1177
<212> PRT
<213> 人工序列-Cry1A.105氨基酸序列(Artificial Sequence)
<400> 39
Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu
1 5 10 15
Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly
20 25 30
Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser
35 40 45
Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile
50 55 60
Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile
65 70 75 80
Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala
85 90 95
Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu
100 105 110
Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu
115 120 125
Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala
130 135 140
Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val
145 150 155 160
Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser
165 170 175
Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg
180 185 190
Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val
195 200 205
Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg
210 215 220
Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val
225 230 235 240
Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro
245 250 255
Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val
260 265 270
Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu
275 280 285
Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr
290 295 300
Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln
305 310 315 320
Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro
325 330 335
Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala
340 345 350
Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg
355 360 365
Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp
370 375 380
Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val
385 390 395 400
Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln
405 410 415
Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His
420 425 430
Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile
435 440 445
Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Glu Phe Asn Asn
450 455 460
Ile Ile Ala Ser Asp Ser Ile Thr Gln Ile Pro Leu Val Lys Ala His
465 470 475 480
Thr Leu Gln Ser Gly Thr Thr Val Val Arg Gly Pro Gly Phe Thr Gly
485 490 495
Gly Asp Ile Leu Arg Arg Thr Ser Gly Gly Pro Phe Ala Tyr Thr Ile
500 505 510
Val Asn Ile Asn Gly Gln Leu Pro Gln Arg Tyr Arg Ala Arg Ile Arg
515 520 525
Tyr Ala Ser Thr Thr Asn Leu Arg Ile Tyr Val Thr Val Ala Gly Glu
530 535 540
Arg Ile Phe Ala Gly Gln Phe Asn Lys Thr Met Asp Thr Gly Asp Pro
545 550 555 560
Leu Thr Phe Gln Ser Phe Ser Tyr Ala Thr Ile Asn Thr Ala Phe Thr
565 570 575
Phe Pro Met Ser Gln Ser Ser Phe Thr Val Gly Ala Asp Thr Phe Ser
580 585 590
Ser Gly Asn Glu Val Tyr Ile Asp Arg Phe Glu Leu Ile Pro Val Thr
595 600 605
Ala Thr Leu Glu Ala Glu Tyr Asn Leu Glu Arg Ala Gln Lys Ala Val
610 615 620
Asn Ala Leu Phe Thr Ser Thr Asn Gln Leu Gly Leu Lys Thr Asn Val
625 630 635 640
Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Thr Tyr Leu Ser
645 650 655
Asp Glu Phe Cys Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val Lys
660 665 670
His Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Ser Asn
675 680 685
Phe Lys Asp Ile Asn Arg Gln Pro Glu Arg Gly Trp Gly Gly Ser Thr
690 695 700
Gly Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val
705 710 715 720
Thr Leu Ser Gly Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln
725 730 735
Lys Ile Asp Glu Ser Lys Leu Lys Ala Phe Thr Arg Tyr Gln Leu Arg
740 745 750
Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Ser Ile Arg Tyr
755 760 765
Asn Ala Lys His Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp
770 775 780
Pro Leu Ser Ala Gln Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg
785 790 795 800
Cys Ala Pro His Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg
805 810 815
Asp Gly Glu Lys Cys Ala His His Ser His His Phe Ser Leu Asp Ile
820 825 830
Asp Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile
835 840 845
Phe Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu
850 855 860
Phe Leu Glu Glu Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val Lys
865 870 875 880
Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu
885 890 895
Thr Asn Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe
900 905 910
Val Asn Ser Gln Tyr Asp Gln Leu Gln Ala Asp Thr Asn Ile Ala Met
915 920 925
Ile His Ala Ala Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr Leu
930 935 940
Pro Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu
945 950 955 960
Leu Glu Gly Arg Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn
965 970 975
Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn Val
980 985 990
Lys Gly His Val Asp Val Glu Glu Gln Asn Asn Gln Arg Ser Val Leu
995 1000 1005
Val Val Pro Glu Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys
1010 1015 1020
Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr
1025 1030 1035 1040
Gly Glu Gly Cys Val Thr Ile His Glu Ile Glu Asn Asn Thr Asp Glu
1045 1050 1055
Leu Lys Phe Ser Asn Cys Val Glu Glu Glu Ile Tyr Pro Asn Asn Thr
1060 1065 1070
Val Thr Cys Asn Asp Tyr Thr Val Asn Gln Glu Glu Tyr Gly Gly Ala
1075 1080 1085
Tyr Thr Ser Arg Asn Arg Gly Tyr Asn Glu Ala Pro Ser Val Pro Ala
1090 1095 1100
Asp Tyr Ala Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg Arg
1105 1110 1115 1120
Glu Asn Pro Cys Glu Phe Asn Arg Gly Tyr Arg Asp Tyr Thr Pro Leu
1125 1130 1135
Pro Val Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp
1140 1145 1150
Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp
1155 1160 1165
Ser Val Glu Leu Leu Leu Met Glu Glu
1170 1175
<210> 40
<211> 2457
<212> DNA
<213> 人工序列-Cry1Ab核苷酸序列(Artificial Sequence)
<400> 40
atggacaaca acccaaacat caacgagtgc atcccgtaca actgcctcag caaccctgag 60
gtcgaggtgc tcggcggtga gcgcatcgag accggttaca cccccatcga catctccctc 120
tccctcacgc agttcctgct cagcgagttc gtgccaggcg ctggcttcgt cctgggcctc 180
gtggacatca tctggggcat ctttggcccc tcccagtggg acgccttcct ggtgcaaatc 240
gagcagctca tcaaccagag gatcgaggag ttcgccagga accaggccat cagccgcctg 300
gagggcctca gcaacctcta ccaaatctac gctgagagct tccgcgagtg ggaggccgac 360
cccactaacc cagctctccg cgaggagatg cgcatccagt tcaacgacat gaacagcgcc 420
ctgaccaccg ccatcccact cttcgccgtc cagaactacc aagtcccgct cctgtccgtg 480
tacgtccagg ccgccaacct gcacctcagc gtgctgaggg acgtcagcgt gtttggccag 540
aggtggggct tcgacgccgc caccatcaac agccgctaca acgacctcac caggctgatc 600
ggcaactaca ccgaccacgc tgtccgctgg tacaacactg gcctggagcg cgtctggggc 660
cctgattcta gagactggat tcgctacaac cagttcaggc gcgagctgac cctcaccgtc 720
ctggacattg tgtccctctt cccgaactac gactcccgca cctacccgat ccgcaccgtg 780
tcccaactga cccgcgaaat ctacaccaac cccgtcctgg agaacttcga cggtagcttc 840
aggggcagcg cccagggcat cgagggctcc atcaggagcc cacacctgat ggacatcctc 900
aacagcatca ctatctacac cgatgcccac cgcggcgagt actactggtc cggccaccag 960
atcatggcct ccccggtcgg cttcagcggc cccgagttta cctttcctct ctacggcacg 1020
atgggcaacg ccgctccaca acaacgcatc gtcgctcagc tgggccaggg cgtctaccgc 1080
accctgagct ccaccctgta ccgcaggccc ttcaacatcg gtatcaacaa ccagcagctg 1140
tccgtcctgg atggcactga gttcgcctac ggcacctcct ccaacctgcc ctccgctgtc 1200
taccgcaaga gcggcacggt ggattccctg gacgagatcc caccacagaa caacaatgtg 1260
ccccccaggc agggtttttc ccacaggctc agccacgtgt ccatgttccg ctccggcttc 1320
agcaactcgt ccgtgagcat catcagagct cctatgttct cctggattca tcgcagcgcg 1380
gagttcaaca atatcattcc gtcctcccaa atcacccaaa tccccctcac caagtccacc 1440
aacctgggca gcggcacctc cgtggtgaag ggcccaggct tcacgggcgg cgacatcctg 1500
cgcaggacct ccccgggcca gatcagcacc ctccgcgtca acatcaccgc tcccctgtcc 1560
cagaggtacc gcgtcaggat tcgctacgct agcaccacca acctgcaatt ccacacctcc 1620
atcgacggca ggccgatcaa tcagggtaac ttctccgcca ccatgtccag cggcagcaac 1680
ctccaatccg gcagcttccg caccgtgggt ttcaccaccc ccttcaactt ctccaacggc 1740
tccagcgttt tcaccctgag cgcccacgtg ttcaattccg gcaatgaggt gtacattgac 1800
cgcattgagt tcgtgccagc cgaggtcacc ttcgaagccg agtacgacct ggagagagcc 1860
cagaaggctg tcaatgagct cttcacgtcc agcaatcaga tcggcctgaa gaccgacgtc 1920
actgactacc acatcgacca agtctccaac ctcgtggagt gcctctccga tgagttctgc 1980
ctcgacgaga agaaggagct gtccgagaag gtgaagcatg ccaagcgtct cagcgacgag 2040
aggaatctcc tccaggaccc caatttccgc ggcatcaaca ggcagctcga ccgcggctgg 2100
cgcggcagca ccgacatcac gatccagggc ggcgacgatg tgttcaagga gaactacgtg 2160
actctcctgg gcactttcga cgagtgctac cctacctact tgtaccagaa gatcgatgag 2220
tccaagctca aggcttacac tcgctaccag ctccgcggct acatcgaaga cagccaagac 2280
ctcgagattt acctgatccg ctacaacgcc aagcacgaga ccgtcaacgt gcccggtact 2340
ggttccctct ggccgctgag cgcccccagc ccgatcggca agtgtgccca ccacagccac 2400
cacttctcct tggacatcga tgtgggctgc accgacctga acgaggactt tcggtag 2457
<210> 41
<211> 1905
<212> DNA
<213> 人工序列-Cry2Ab核苷酸序列(Artificial Sequence)
<400> 41
atggacaact ccgtcctgaa ctctggtcgc accaccatct gcgacgccta caacgtcgcg 60
gcgcatgatc cattcagctt ccagcacaag agcctcgaca ctgttcagaa ggagtggacg 120
gagtggaaga agaacaacca cagcctgtac ctggacccca tcgtcggcac ggtggccagc 180
ttccttctca agaaggtcgg ctctctcgtc gggaagcgca tcctctcgga actccgcaac 240
ctgatctttc catctggctc caccaacctc atgcaagaca tcctcaggga gaccgagaag 300
tttctcaacc agcgcctcaa cactgatacc cttgctcgcg tcaacgctga gctgacgggt 360
ctgcaagcaa acgtggagga gttcaaccgc caagtggaca acttcctcaa ccccaaccgc 420
aatgcggtgc ctctgtccat cacttcttcc gtgaacacca tgcaacaact gttcctcaac 480
cgcttgcctc agttccagat gcaaggctac cagctgctcc tgctgccact ctttgctcag 540
gctgccaacc tgcacctctc cttcattcgt gacgtgatcc tcaacgctga cgagtggggc 600
atctctgcag ccacgctgag gacctaccgc gactacctga agaactacac cagggactac 660
tccaactatt gcatcaacac ctaccagtcg gccttcaagg gcctcaatac gaggcttcac 720
gacatgctgg agttcaggac ctacatgttc ctgaacgtgt tcgagtacgt cagcatctgg 780
tcgctcttca agtaccagag cctgctggtg tccagcggcg ccaacctcta cgccagcggc 840
tctggtcccc aacaaactca gagcttcacc agccaggact ggccattcct gtattcgttg 900
ttccaagtca actccaacta cgtcctcaac ggcttctctg gtgctcgcct ctccaacacc 960
ttccccaaca ttgttggcct ccccggctcc accacaactc atgctctgct tgctgccaga 1020
gtgaactact ccggcggcat ctcgagcggc gacattggtg catcgccgtt caaccagaac 1080
ttcaactgct ccaccttcct gccgccgctg ctcaccccgt tcgtgaggtc ctggctcgac 1140
agcggctccg accgcgaggg cgtggccacc gtcaccaact ggcaaaccga gtccttcgag 1200
accacccttg gcctccggag cggcgccttc acggcgcgtg gaaattctaa ctacttcccc 1260
gactacttca tcaggaacat ctctggtgtt cctctcgtcg tccgcaacga ggacctccgc 1320
cgtccactgc actacaacga gatcaggaac atcgcctctc cgtccgggac gcccggaggt 1380
gcaagggcgt acatggtgag cgtccataac aggaagaaca acatccacgc tgtgcatgag 1440
aacggctcca tgatccacct ggcgcccaat gattacaccg gcttcaccat ctctccaatc 1500
cacgccaccc aagtgaacaa ccagacacgc accttcatct ccgagaagtt cggcaaccag 1560
ggcgactccc tgaggttcga gcagaacaac accaccgcca ggtacaccct gcgcggcaac 1620
ggcaacagct acaacctgta cctgcgcgtc agctccattg gcaactccac catcagggtc 1680
accatcaacg ggagggtgta cacagccacc aatgtgaaca cgacgaccaa caatgatggc 1740
gtcaacgaca acggcgcccg cttcagcgac atcaacattg gcaacgtggt ggccagcagc 1800
aactccgacg tcccgctgga catcaacgtg accctgaact ctggcaccca gttcgacctc 1860
atgaacatca tgctggtgcc aactaacatc tcgccgctgt actga 1905
<210> 42
<211> 3534
<212> DNA
<213> 人工序列-Cry1A.105核苷酸序列(Artificial Sequence)
<400> 42
atggacaaca acccaaacat caacgagtgc atcccgtaca actgcctcag caaccctgag 60
gtcgaggtgc tcggcggtga gcgcatcgag accggttaca cccccatcga catctccctc 120
tccctcacgc agttcctgct cagcgagttc gtgccaggcg ctggcttcgt cctgggcctc 180
gtggacatca tctggggcat ctttggcccc tcccagtggg acgccttcct ggtgcaaatc 240
gagcagctca tcaaccagag gatcgaggag ttcgccagga accaggccat cagccgcctg 300
gagggcctca gcaacctcta ccaaatctac gctgagagct tccgcgagtg ggaggccgac 360
cccactaacc cagctctccg cgaggagatg cgcatccagt tcaacgacat gaacagcgcc 420
ctgaccaccg ccatcccact cttcgccgtc cagaactacc aagtcccgct cctgtccgtg 480
tacgtccagg ccgccaacct gcacctcagc gtgctgaggg acgtcagcgt gtttggccag 540
aggtggggct tcgacgccgc caccatcaac agccgctaca acgacctcac caggctgatc 600
ggcaactaca ccgaccacgc tgtccgctgg tacaacactg gcctggagcg cgtctggggc 660
cctgattcta gagactggat tcgctacaac cagttcaggc gcgagctgac cctcaccgtc 720
ctggacattg tgtccctctt cccgaactac gactcccgca cctacccgat ccgcaccgtg 780
tcccaactga cccgcgaaat ctacaccaac cccgtcctgg agaacttcga cggtagcttc 840
aggggcagcg cccagggcat cgagggctcc atcaggagcc cacacctgat ggacatcctc 900
aacagcatca ctatctacac cgatgcccac cgcggcgagt actactggtc cggccaccag 960
atcatggcct ccccggtcgg cttcagcggc cccgagttta cctttcctct ctacggcacg 1020
atgggcaacg ccgctccaca acaacgcatc gtcgctcagc tgggccaggg cgtctaccgc 1080
accctgagct ccaccctgta ccgcaggccc ttcaacatcg gtatcaacaa ccagcagctg 1140
tccgtcctgg atggcactga gttcgcctac ggcacctcct ccaacctgcc ctccgctgtc 1200
taccgcaaga gcggcacggt ggattccctg gacgagatcc caccacagaa caacaatgtg 1260
ccccccaggc agggtttttc ccacaggctc agccacgtgt ccatgttccg ctccggcttc 1320
agcaactcgt ccgtgagcat catcagagct cctatgttct cttggataca ccgtagtgct 1380
gagttcaaca acatcattgc atccgacagc attactcaaa tacccttggt gaaagcacat 1440
acacttcagt caggtactac tgttgtcaga ggtccagggt ttacaggagg agacattctt 1500
cgtcgcacaa gtggaggacc ctttgcttac actattgtta acatcaatgg ccaattgccc 1560
caaaggtatc gtgcaagaat ccgctatgcc tctactacaa atctcaggat ctacgtgact 1620
gttgcaggtg aaaggatctt tgctggtcag ttcaacaaga ctatggatac cggtgaccct 1680
ttgacattcc aatcttttag ctacgcaact atcaacacag cttttacatt cccaatgagc 1740
cagagtagct tcacagtagg tgctgacact ttcagctcag ggaatgaagt ttacatcgac 1800
aggtttgaat tgattccagt tactgcaacc ctcgaggctg agtacaacct tgagagagcc 1860
cagaaggctg tgaacgccct ctttacctcc accaatcagc ttggcttgaa aactaacgtt 1920
actgactatc acattgacca agtgtccaac ttggtcacct accttagcga tgagttctgc 1980
ctcgacgaga agcgtgaact ctccgagaaa gttaaacacg ccaagcgtct cagcgacgag 2040
aggaatctct tgcaagactc caacttcaaa gacatcaaca ggcagccaga acgtggttgg 2100
ggtggaagca ccgggatcac catccaagga ggcgacgatg tgttcaagga gaactacgtc 2160
accctctccg gaactttcga cgagtgctac cctacctact tgtaccagaa gatcgatgag 2220
tccaaactca aagccttcac caggtatcaa cttagaggct acatcgaaga cagccaagac 2280
cttgaaatct actcgatcag gtacaatgcc aagcacgaga ccgtgaatgt cccaggtact 2340
ggttccctct ggccactttc tgcccaatct cccattggga agtgtggaga gcctaacaga 2400
tgcgctccac accttgagtg gaatcctgac ttggactgct cctgcaggga tggcgagaag 2460
tgtgcccacc attctcatca cttctccttg gacatcgatg tgggatgtac tgacctgaat 2520
gaggacctcg gagtctgggt catcttcaag atcaagaccc aagacggaca cgcaagactt 2580
ggcaaccttg agtttctcga agagaaacca ttggtcggtg aagctctcgc tcgtgtgaag 2640
agagcagaga agaagtggag ggacaaacgt gagaaactcg aatgggaaac taacatcgtt 2700
tacaaggagg ccaaagagtc cgtggatgct ttgttcgtga actcccaata tgatcagttg 2760
caagccgaca ccaacatcgc catgatccac gccgcagaca aacgtgtgca cagcattcgt 2820
gaggcttact tgcctgagtt gtccgtgatc cctggtgtga acgctgccat cttcgaggaa 2880
cttgagggac gtatctttac cgcattctcc ttgtacgatg ccagaaacgt catcaagaac 2940
ggtgacttca acaatggcct cagctgctgg aatgtgaaag gtcatgtgga cgtggaggaa 3000
cagaacaatc agcgttccgt cctggttgtg cctgagtggg aagctgaagt gtcccaagag 3060
gttagagtct gtccaggtag aggctacatt ctccgtgtga ccgcttacaa ggagggatac 3120
ggtgagggtt gcgtgaccat ccacgagatc gagaacaaca ccgacgagct taagttctcc 3180
aactgcgtcg aggaagaaat ctatcccaac aacaccgtta cttgcaacga ctacactgtg 3240
aatcaggaag agtacggagg tgcctacact agccgtaaca gaggttacaa cgaagctcct 3300
tccgttcctg ctgactatgc ctccgtgtac gaggagaaat cctacacaga tggcagacgt 3360
gagaaccctt gcgagttcaa cagaggttac agggactaca caccacttcc agttggctat 3420
gttaccaagg agcttgagta ctttcctgag accgacaaag tgtggatcga gatcggtgaa 3480
accgagggaa ccttcatcgt ggacagcgtg gagcttctct tgatggagga ataa 3534
<210> 43
<211> 888
<212> DNA
<213> 人工序列-ACe1_3转基因植物核苷酸序列(Artificial Sequence)
<400> 43
atgtcctcta cagatgttca ggagcggctg cgggacctag cgcgtgaaaa tgaagcaggc 60
acgctgaacg aggcgtggaa cacaaatttc aagcctagtg acgagcagca gttttcttat 120
tctcctaccg agggcatcgt cttcctcacg ccgccgaaga atgtcatcgg ggagcgacgc 180
atctcacagt acaaggtgaa taatgcatgg gctacacttg aaggctcacc aactgaagtg 240
agcggtacgc cgctttatgc agggaagaac gtcttggata attccaaggg caccatggat 300
caagagctat taactcccga gttcaactac acatacactg aatcaacgtc caacaccacg 360
acccacggcc tcaagctcgg agtgaaaact acagccacga tgaagtttcc aattgcgcag 420
ggctccatgg aggcttccac cgagtacaac ttccagaact cgtcaacgga caccaagaca 480
aagcaggttt cttacaaatc tccttcgcag aagataaaag ttccggctgg caaaaccttc 540
cgcgtgctgg cgtacctgaa tactggaagc atcagcgggg aagccaattt gtacgccaac 600
gttggaggga tagcctgggg tgtattgccc ggatatccaa atggtggcgg cgtcaacatt 660
ggtgctgtgc tcactaaatg ccagcaaaag ggatggggcg actttagaaa ttttcaacca 720
agtggaaggg atgtcattgt gaaagggcaa ggaacattca ccagcaacta tggtaccgac 780
ttcatcctga agattgagga cattactgat agtaaactta ggaataacaa cggttcgggg 840
actgtagtgc aagagatcaa ggttcccctc ataagaacag aaatatag 888
<210> 44
<211> 888
<212> DNA
<213> 人工序列-ACe1_4转基因植物核苷酸序列(Artificial Sequence)
<400> 44
atgagttcca cagatgttca agaacgccta cgcgacctcg ccagggaaaa tgaagcaggc 60
accttgaatg aggcgtggaa caccaatttc aagccatctg atgagcagca gtttagctac 120
agcccaacgg agggcatcgt cttcctcact cctcccaaga acgtcattgg agagcggaga 180
atatcacagt acaaggtgaa caatgcatgg gcgacgctgg aaggatcacc tacagaagta 240
agcggtactc cattatacgc tggcaagaat gttttggaca acagtaaagg cacgatggac 300
caggagctgc taacgccgga gttcaactat acctacacag aatccacttc aaatacaaca 360
acgcacggct tgaagctcgg tgtcaagacc accgccacca tgaagttccc catcgcgcag 420
gggtccatgg aggcttctac cgagtataac tttcaaaatt cgtccacaga tacaaaaact 480
aagcaggtgt cgtacaaatc tccgtcacaa aaaataaaag ttcccgcggg aaaaacattt 540
cgcgtgctgg catatcttaa tactggaagc atttctgggg aggccaacct gtacgccaat 600
gttggtggta ttgcttgggg cgtgcttcct ggttatccaa atggcggcgg gatcaacatt 660
ggcgccgtcc tcaccaagtg ccaacagaag gggtggggtg acttcaggaa cttccagccg 720
tcggggcgtg atgtcattgt gaagggccaa ggaacattca aatcaaacta cggcactgat 780
tttattctca agatcgagga catcaccgac tccaagctga gaaataacaa cgggagtgga 840
acggtagtgc aggaaatcaa ggttcctctt atacggactg agatctag 888
<210> 45
<211> 888
<212> DNA
<213> 人工序列-ACe1_5转基因植物核苷酸序列(Artificial Sequence)
<400> 45
atgagctcta ctgatgtcca ggaacgatta cgcgacctcg cgcgtgagaa tgaagccggg 60
acgctgaacg aggcgtggaa cacaaacttc aagccgtccg atgagcagca gttttcttac 120
agccccactg aaggcatcgt cttcttgaca ccccctaaga atgtcattgg cgagcggcgc 180
atatctcagt ataaagtgtc gaacgcctgg gcaacattgg aggggagtcc aacagaggtg 240
tcgggcacac ctctttatgc cgggaagaac gtgctggata attccaaggg gactatggac 300
caagaacttt taaccccgga gttcaactac acctacactg agtctacctc aaacacgaca 360
actcacgggc taaagcttgg tgtaaaaact acggcgacaa tgaagtttcc aatagcgcaa 420
ggctcgatgg aggcaagcac cgagtacaat ttccagaatt catctaccga cactaaaacc 480
aagcaggtta gctacaagtc accatcccag aagatcaaag tccccgccgg aaaaacgtat 540
agagttcttg cttatctgaa cactggcagt atcagtggag aagctaactt gtacgctaat 600
gttggcggca ttgcatgggg agttctcccg ggatacccaa atggcggtgg tattaacatc 660
ggtgctgtgc tcacgaagtg ccaacaaaag ggctggggtg attttaggaa ttttcagccg 720
tcaggacggg acgttattgt gaaggggcaa ggtaccttca aatccaacta tggcaccgac 780
ttcattctca agattgaaga catcacggat tccaaactga gaaataacaa tggaagcggt 840
acggtagtgc aggagatcaa ggtccctctg ataaggacag aaatatag 888
<210> 46
<211> 888
<212> DNA
<213> 人工序列-ACe1_6转基因植物核苷酸序列(Artificial Sequence)
<400> 46
atgtccagta cagatgtgca agaaagattg cgtgatctag ctagagaaaa tgaagctggg 60
acgctgaatg tagcatggaa cacaaatttc aagccctctg atgagcagca gttctcatat 120
agtcctacag aaggtattgt ttttttaaca ccaccaaaga acgtcattgg cgagcggcga 180
atcagccagt acaaggtgaa caacgcatgg gctacccttg agggcagtcc tactgaggtg 240
agcggcacgc ccctttatgc gggcaagaat gtcctggaca actccaaagg aacaatggac 300
caggagctcc tcactccaga gttcaactac acctacacag aaagcacttc aaataccacg 360
acgcacggcc tgaaacttgg tgttaagaca actgccacca tgaagtttcc tatcgcgcag 420
ggcagcatgg aggcgtcgac cgagtacaac tttcaaaatt cttctactga cacaaaaaca 480
aagcaggtct cgtacaaatc accatcacag aaaataaagg tccccgccgg caaaaccttc 540
cgcgtgctgg catatcttaa cactgggtcc atctccgggg aggccaacct ctacgccaat 600
gttggaggga ttgcttgggg tgttttgcct ggttacccga atgggggcgg catcaacatt 660
ggtgccgtac tcaccaagtg ccaacaaaag ggatggggcg acttccggaa cttccagccg 720
tcggggaggg atgttattgt gaagggccaa ggaacgttta agagcaacta tggtactgat 780
ttcatcctca agatagagga catcacggac tccaaattgc gcaataataa tggatctgga 840
accgtcgtgc aggaaatcaa ggtgccgctg ataaggacgg agatatag 888
<210> 47
<211> 888
<212> DNA
<213> 人工序列-ACe1_8转基因植物核苷酸序列(Artificial Sequence)
<400> 47
atgtccagta ctgatgtgca agagaggctc cgcgacctag cccgggaaaa tgaagctggc 60
acgctcaacg tagcttggaa cacaaatttc aagccgtccg acgagcagca gttttcgtac 120
tcccctactg aaggaattgt gtttcttacg ccgcccaaga acgttattgg tgaaagaagg 180
atttctcagt acaaggtcaa taatgcgtgg gcgacgctgg aggggtctcc aacagaagtt 240
tcaggaactc ccctctatgc aggcaagaat gtgctcgaca acagcaaggg cacaatggat 300
caggagcttt tgacacctga gttcaactac acctacacgg agtctacatc aaatacaact 360
acgcacgggc tgaagctcgg cgttaagacc accgccacca tgaagttccc catcgcgcag 420
ggctccatgg aggcgtcgac tgagtacaac ttccagaact ccagcactga caccaaaact 480
aagcaggtgt cgtataagag cccatctcaa aagatcaagg ttcctgctgg gaagacatat 540
agagtcctag catatctgaa caccggcagc ataagcgggg aggccaacct gtacgccaat 600
gttggtggta ttgcatgggg cgtccttccg ggctacccaa atggaggcgg catcaacatc 660
ggcgccgtct taacaaaatg ccaacagaag gggtggggtg attttcgaaa ttttcaacca 720
agtggacgtg atgtcatagt aaaaggacaa ggtaccttca aatcaaatta tgggacggac 780
ttcatcttga aaattgaaga catcacagat tcaaaattgc gcaacaacaa tggtagtgga 840
acggtggtgc aggaaataaa agtgccgctg attcggaccg agatctag 888
<210> 48
<211> 888
<212> DNA
<213> 人工序列-ACe1_9转基因植物核苷酸序列(Artificial Sequence)
<400> 48
atgtcatcta ctgatgttca ggagaggcta agggatcttg caagagaaaa tgaagctggt 60
acgctcaatg tggcgtggaa cacaaatttc aagccatctg atgagcagca gttctcctat 120
tctccaacag aaggtattgt attccttacg ccgcccaaga atgtcatagg tgaacgtcgg 180
atttcacagt acaaggtcaa caatgcatgg gcgacgctgg aaggaagtcc taccgaggtg 240
tccggaactc ctctgtacgc cggcaagaac gtccttgaca acagcaaggg tacaagtgat 300
caagaacttt taacacctga gttcaactac acctatacag agtcgacgag caacaccacc 360
acgcacggcc tcaagctggg ggtgaaaact accgccacca tgaagtttcc aattgctcaa 420
ggatccatgg aggcctcgac ggagtacaac ttccagaatt catcgaccga cacaaaaaca 480
aagcaggtta gctacaaaag cccgtctcag aagatcaaag ttcctgctgg gaagacttat 540
agggtgctgg cgtacctcaa cactggcagc atatccggcg aggcgaattt gtatgcaaat 600
gttggaggca tcgcttgggg tgtgctcccc ggctacccaa atggcggcgg gatcaacatc 660
ggcgccgtcc tcaccaagtg ccagcaaaag ggatggggcg acttccgcaa ctttcagccg 720
tcggggcgcg acgtcatcgt gaaggggcaa ggcaccttta aatcaaatta tggaacagat 780
tttatactga aaattgagga catcacagac tccaaattgc ggaataacaa cggtagtggt 840
actgttgtac aagagatcaa ggtgcccttg attagaactg aaatatag 888
<210> 49
<211> 888
<212> DNA
<213> 人工序列-ACe1_10转基因植物核苷酸序列(Artificial Sequence)
<400> 49
atgtccagca cggatgtcca agaacgtctg cgcgacttgg cacgcgagaa tgaggcgggg 60
accctaaacg aggcctggca cacaaatttc aagccctccg acgaacaaca gttctcctat 120
agcccgacgg aaggcatcgt gttcctcact ccaccaaaga acgtaatagg cgagaggaga 180
atttctcaat ataaggtgaa caatgcatgg gctacactgg agggcagccc aaccgaggtc 240
tcaggcaccc cgctctacgc ggggaaaaat gtgctcgaca actctaaagg aacaatggac 300
caggagcttc tgacgccgga gttcaactac acgtacacag aaagtactag taacacgatt 360
acccatggat tgaagctcgg agtgaaaact actgctacta tgaagttccc catcgcccaa 420
ggctccatgg aggcctctac ggagtacaac ttccagaatt cgtctaccga caccaagaca 480
aagcaggtct cgtacaagtc accgagccag aagattaaag ttcctgcagg caagacattt 540
agggtcctcg cgtacctgaa cactggtagc atttctggtg aagctaacct ttatgctaat 600
gttggaggga tcgcgtgggg cgttcttcct ggttatccta atggtggcgg cgtgaacatt 660
ggagccgttt tgacaaaatg ccagcagaaa gggtggggtg actttaggaa ttttcagccc 720
agtgggcggg atgtaattgt caagggccag ggtacctttg agtcgaatta tggaacagat 780
ttcatattaa agatcgaaga tatcactgat tcaaagctgc ggaataacaa cgggtcagga 840
accgtggtgc aagagatcaa agttcctctt atcagaacgg aaatatag 888
<210> 50
<211> 888
<212> DNA
<213> 人工序列-ACe1_11转基因植物核苷酸序列(Artificial Sequence)
<400> 50
atgagctcaa cagatgtgca ggagcggctg cgcgatctgg cgcgtgaaaa tgaggctgga 60
accttgaatg aagcatggaa caccaacttc aagccatcgg acgagcagca gttttcatac 120
agcccgacgg aaggtattgt tttcctaacg ccgcccaaga acgtcattgg agaacggcgc 180
atctctcaat acaaggtgaa caacgcctgg gcaactcttg agggctcgcc tactgagatg 240
tcagggactc cgctctatgc tgggaagaat gtgctggaca actcaaaagg aacttctgat 300
caagaattac ttacaccaga gttcaactac acctatacgg aaagtacgtc caacacaacc 360
acacacggcc tcaagttagg tgtgaagacg acggcgacca tgaagttccc catagcacag 420
gggagcatgg aggcgtcgac agagtacaac ttccagaata gttcgactga tacaaaaaca 480
aagcaggtga gctacaaatc tcctagtcaa aagataaaag ttcctgctgg caaaactttt 540
agagtactcg cctaccttaa tactggatcc atctccggcg aggccaactt gtatgctaat 600
gtcggcggga tcgcgtgggg cgtgctcccc gggtacccta atggtggtgg cgtcaacatc 660
ggcgccgtcc tcaccaagtg ccagcagaag ggctggggtg acttcaggaa ttttcagcca 720
tctggaagag acgttattgt caaaggtcaa ggaaccttca cttccaatta tggcaccgat 780
tttatattga aaattgagga catcacggac tccaagctga ggaacaataa tggcagcggg 840
actgtagttc aagagatcaa ggttccgctg atccgaacag aaatttag 888
<210> 51
<211> 888
<212> DNA
<213> 人工序列-ACe1_12转基因植物核苷酸序列(Artificial Sequence)
<400> 51
atgtcctcca ccgacgtgca agagcgcctc cgggaccttg ccagagagga tgaggctggc 60
acctttaatg aagcatggaa cacaaatttc aaaccaagcg acgagcaaca attttcttat 120
tctccgacgg agggaatcgt cttcctcaca cctccaaaaa acgtgattgg cgaacgacgg 180
atatctcagt acaaggttaa taatgcctgg gccacgctgg aaggctctcc taccgaggtt 240
tcagggactc cgctatatgt tggtaagaac gtcctggaca actcgaaggg gacctcggat 300
caggagctcc taacaccgga gttcaactac acgtatactg aaagtaccag taacacgacc 360
acccacgggc ttaagttggg cgtgaaaact actgcaacga tgaagttccc catcgcgcag 420
ggaagcatgg aggctagcac tgagtacaac ttccaggata gctccacaga cacaaccact 480
aaaaccgtca gctacaaatc accctcacaa aaaatcaagg tgcccgcggg caagactttc 540
cgcgtattgg cgtacctgaa tacaggaagt atttcgggag aggccaacct ctatgcaaat 600
gtgggcggga tagcttgggg tgttcttcct ggatacccaa atggaggggg tgtaaacatc 660
ggcgcggtgc tgaccaagtg ccagcagaag ggctggggcg agtttaggaa ttttcagcca 720
tccgggcgtg atgtaattgt caagggtcaa ggcacattta agtcaaatta tggtacggac 780
ttcattttga agatcgaaga tataactgat tccaagttaa ggaacaacaa tggctccggg 840
acggtcgtgc aggaaattaa agttccgctg atcagaacag aaatatag 888
<210> 52
<211> 888
<212> DNA
<213> 人工序列-ACe1_13转基因植物核苷酸序列(Artificial Sequence)
<400> 52
atgtcatcta ctgatgttca agaacgcctc cgtgatttag cgcgcgagaa tgaagctggg 60
actttgaatg tagcttggaa cactaatttc aagccttcag atgagcagca attttcttat 120
tctccaacag aagggatcgt cttcctcacc ccgcccaaga acgtcatcgg cgagaggagg 180
atatcacagt acaaagtcaa caacgcgtgg gcgacgctgg aaggaagtcc cactgaggtg 240
tccggcacac cactatatgc agggaaaaat gttctggaca attccaaagg aacatctgat 300
caagagctgc tcacgccgga gttcaactac acatacactg agagcactag caacaccacg 360
acgcacggcc tcaagctagg cgtcaagacc acagccacca tgaaatttcc aattgctcag 420
ggctccatgg aggcatcgac ggagtacaac ttccagaaca gctcgactga caccaagacg 480
aagcaggttt cttacaagag tccttcccaa aagataaaag tgccggcggg taagacattt 540
cgggtgctgg cctacctcaa caccgggagc atctcggggg aggccaatct ttatgccaac 600
gtcggcggaa ttgcatgggg cgtgttgccg ggctacccaa atggaggtgg catcaacatt 660
ggtgctgtat tgacaaaatg ccagcagaag gggtggggag atttcagaaa ttttcagccc 720
tcaggtagag acgtgattgt gaagggccaa ggaaccttca aatcaaatta tggtaccgac 780
ttcatcctga agattgaaga cataacagac agcaaacttc ggaacaataa tggcagtggt 840
acggttgtgc aggagatcaa ggttcctctt atccgaactg aaatttag 888
<210> 53
<211> 888
<212> DNA
<213> 人工序列-ACe1_14转基因植物核苷酸序列(Artificial Sequence)
<400> 53
atgagttcta ctgatgtgca ggaaaggttg cgtgacttag ctagagaaaa tgaagccggc 60
accctaaatg aggcatggaa tactaatttc aagccctctg atgagcaaca gttctcttac 120
agcccaactg aggggatcgt cttcttgact cctccaaaga acgtcatagg agaacgacgc 180
atttcacagt acaaggttaa taatgcttgg gcgacgctgg aaggtagccc caccgagatg 240
tcgggtacac ctctttatgc tgggaaaaat gttctggaca actcgaaagg aacaagtgat 300
caggagctcc tgacgccgga gttcacatat acgtacactg aatcaacctc aaacacaaca 360
acccacggcc tgaaacttgg cgtcaaaaca acagccacca tgaagtttcc tatagcacag 420
ggctccatgg aggcgagcac cgagtacaac ttccagaatt cttccactga caccaaaact 480
aagcaagttt cttataagag cccttcacaa aagatcaagg tcccggcggg caagacattt 540
agagttcttg catatctcaa caccgggtcc ataagcgggg aggctaacct gtacgccaac 600
gtgggcggca tcgcgtgggg agtattgccc ggctacccaa atggtggtgg tgtgaacatc 660
ggcgccgtct taacgaagtg ccagcagaag ggctggggcg actttaggaa ctttcagcca 720
tcggggaggg acgttatagt gaaggggcaa ggtacgttca cttccaatta tggaaccgac 780
ttcatcctca aaattgaaga tattactgat tccaagctcc gcaacaacaa tggaagtggt 840
acggtggtgc aagagattaa agtaccgctc atccggacgg agatttag 888
<210> 54
<211> 888
<212> DNA
<213> 人工序列-ACe1_15转基因植物核苷酸序列(Artificial Sequence)
<400> 54
atgtcgtcca ctgatgttca agagaggctt cgggacctgg cgcgggagaa cgaggcgggc 60
accctcaacg aagcttggaa cactaatttt aagccttccg atgagcagca attctcatat 120
tcaccgaccg aaggcatcgt gttcttgact ccaccaaaga atgttattgg ggagaggcga 180
atctcgcagt acaaggtgaa caacgcgtgg gcgacgctgg aaggttctcc gacggaggtc 240
agtggaaccc ccctttacgc cggtaagaac gttcttgaca attcaaaagg aacatcagat 300
caggaactgc ttacgcccga gttcaactac acttatactg aaagcacaag caacacgaca 360
acacacggac tgaagctggg cgtgaagacg accgctacca tgaagtttcc aattgctcaa 420
ggatctatgg aggcaagtac agagtacaac tttcaggaca gctccaccga tacaaccact 480
aaaacagtct cctataaatc tccctcgcag aagatcaagg tcccggcagg gaaaacattt 540
cgtgtgctag cctacttgaa caccgggagc atatctggtg aagcaaattt gtatgccaat 600
gtaggtggta tagcttgggg cgtattacct ggctacccga atggcggcgg ggtcaacata 660
ggcgccgtgc tcacaaaatg ccagcagaaa ggctggggtg atttcagaaa tttccaacct 720
tctgggagag acgtgatcgt caagggccag ggcactttca agagcaatta tggaactgac 780
ttcattctga agattgagga catcaccgat agtaagctcc gcaacaataa tgggtcaggg 840
acggtggttc aagaaataaa agttccactc atccgcactg agatttag 888
<210> 55
<211> 888
<212> DNA
<213> 人工序列-ACe1_16转基因植物核苷酸序列(Artificial Sequence)
<400> 55
atgtcttcta ctgatgttca agagcgcctc cgagacttgg ctagggaaaa tgaagccggc 60
actcttaacg tggcgtggaa cacaaatttc aagcctagtg atgagcagca gttctcgtat 120
agcccgactg agggcttcat ttttttaacg ccgccgaaga atgtcattgg agagcgtcgg 180
atctcccact ataaagtcaa taacgcctgg gcgacgctgg aagggtcccc cacagaggtg 240
tcaggtaccc ctctgtacgc cggcaggaat gttttggaca acagcaaggg caccatcgac 300
caggagatgc tgacgccaga gttcaactac acctacacag aaggaacttc aaacacaacg 360
acccatgggc tgaaactcgg ggtgaagact acagccacca tgaagtttcc tattgcgcag 420
ggcagcatgg aggcatcaac ggagtacaac ttccagaact catctacaga tacaaaaact 480
aagcaagtaa gctacaagag tccttctcag aagatcaagg tgcccgcggg caagaccttt 540
cgggtgctgg catacctaaa cacaggatcc atcagtggtg aagctaatct ctatgctaac 600
gtcggtgggg ttgcttgggg cgtccttccc ggctacccaa atggaggcgg ggtaaatatt 660
ggcgcagtgc tcaccaaatg ccagcaaaaa gggtggggag acttcagaaa ctttcagccg 720
agcggaagag atgtcatcgt caaggggcaa ggaacgttca cttcgaatta tggtacggat 780
tttatattga agattgagga cataaccgac tcgaaacttc gcaacaataa tggttccggc 840
acagtggttc aagaaatcaa agttccactc atcaggactg aaatatag 888
<210> 56
<211> 888
<212> DNA
<213> 人工序列-ACe1_17转基因植物核苷酸序列(Artificial Sequence)
<400> 56
atgagctcaa ctgacgtgca ggaaaggctg cgggacctcg ccagggagga cgaggccggc 60
acgtttaatg aagcgtggaa cgcaaatttc aaaccttctg atgagccgca gttctccaat 120
tcaccaacac aaggtatagt gatacttgct cccccaaaga acgtcattgg tgaaagaaga 180
atatcccagt acaaagtcaa taatgcctgg gcgacgctgg aggggtctcc gacggaggtc 240
tcgggcactc cactatacgc ggggcgcaac gtgctggaca attctaaagg caccatggac 300
caagagcttt tgacgccgga gttcacctac acatatactg agagcacctc taacacaacg 360
acgcacggct taaaactggg agtgaagaca actgcaacaa tgaagtttcc tattgctcag 420
ggcagcatgg aggcaagtac tgaatataat tttcaaaaca gctcgacaga tacaaaaaca 480
aagcaggtta gctacaagtc cccatcgcag aagatcaagg tgcctgctgg caagaccttc 540
cgcgtattgg cctacctcaa caccggatct attagtggag aagccaacct ttatgctaat 600
gttggtggaa ttgcatggcg tgtttcaccg ggctacccca acggaggtgg cgtgaatatc 660
ggcgcggtcc taaccaagtg ccagcagaaa gggtggggcg acttccggaa cttccagccc 720
tcggggcgcg atgtaatagt caaggggcaa gggacgttca agagtgatta tggaactgat 780
tttatcctca agattgaaga catcaccgat tcaaagtcca ggaacaacaa tggatcaggt 840
actgttgtgc aagaaattaa agttcctctc atccgaaccg agatctag 888
<210> 57
<211> 888
<212> DNA
<213> 人工序列-ACe1_18转基因植物核苷酸序列(Artificial Sequence)
<400> 57
atgagctcta ccgacgtgca agagagactc cgtgacctgg cccgggaaaa tgaagctggc 60
actcttaatg tagcgtggaa cacgaacttc aagccttcag atgaacagca gttcagttac 120
agccctactg aagggttcat ctttctgacg ccccccaaga acgtaatcgg cgagcgcagg 180
attagccact acaaggtgaa taacgcctgg gcgacattgg agggatcacc aacggaagtt 240
tcgggtacgc ccctgtatgc tgggaggaat gttctcgaca atagcaaggg gaccattgat 300
caagagatgt taaccccgga gtttaactac acctacactg agggcacttc aaatacaaca 360
actcatggtc tcaaattggg tgtcaagaca acagccacca tgaagttccc aatagcccag 420
ggctccatgg aggcgagcac agagtacaat tttcagaatt cttccactga cacgaagacc 480
aagcaagtct cctacaaatc tccgtcccag aagatcaaag tcccggctgg aaaaacattt 540
cgagtcctcg cgtaccttaa cacggggtcg atctcaggtg aagcaaatct atatgcaaac 600
gttgggggcg tggcatgggg tgttctacct ggctatccag atggtggcgg agtgaacata 660
ggcgccgtcc ttactaaatg ccagcaaaaa ggatggggcg acttcaggaa cttccagcct 720
tctgggcggg acgtcattgt gaagggccaa ggaacgttca ccagtaacta tggaactgat 780
tttatattga agattgagga tattacagat agtaaactgc gcaacaacaa tggttcgggc 840
accgtggtgc aggagatcaa ggttccgctg atcagaacag aaatatag 888
<210> 58
<211> 888
<212> DNA
<213> 人工序列-ACe1_19转基因植物核苷酸序列(Artificial Sequence)
<400> 58
atgagcagta cagatgttca ggagcgcttg agggatcttg cgcgggagaa cgaggctgga 60
accttgaatg aagcatggaa caccaacttt aagccatcag atgagcagca gttctcctat 120
tctcccacgg aaggaattgt cttcctcacg ccgcccaaga acgtcatcgg cgagagacgg 180
atatctcaat ataaggtgaa taatgcttgg gcgacgttgg aaggctcacc tacagaggtc 240
tctggaacac cgctttatgc cgggaaaaat gtactagaca actctaaagg aacaatggat 300
caagaactgc tgactcctga gttcaactat acctacacag aatcaacatc caacactacg 360
acgcacggtc tcaaagtcgg cgtgaagacg accgcgacca tgaagtttcc catagcacag 420
gggagcatgg aggcctccac tgagtacaat ttccaggact cgtcaactga cacgactaca 480
aagactgtat cctacaagag tccatcccag aagatcaagg ttcctgctgg caaaacattt 540
cgcgtgcttg catacctcaa tactggaagc atttctgggg aagccaacct gtatgctaat 600
gttggtggta tcgcgtgggg tgttctgccg ggctacccaa atgggggggg cgttaacatt 660
ggcgccgtgt taaccaagtg ccagcaaaaa ggctggggcg actttagaaa tttccaacca 720
tcgggccgag atgtgatcgt caaggggcag ggtaccttca agagcaacta cggcactgat 780
ttcattctca aaattgagga catcaccgac agcaagctgc gtaataacaa tggttcggga 840
actgtggtgc aagagattaa agtcccgctc atcaggaccg aaatatag 888
<210> 59
<211> 888
<212> DNA
<213> 人工序列-ACe1_20转基因植物核苷酸序列(Artificial Sequence)
<400> 59
atgtcatcaa ctgatgtaca agagaggctg cgagacctgg ctcgggaaaa cgaggctggg 60
accctcaatg aggcctggaa caccaatttt aaaccgagcg atgagcagca gttctcgtat 120
tcccccactg aagggatagt gttcctgact cctccgaaga atgtaatcgg cgagaggaga 180
atctcacaat acaaggtcaa taatgcgtgg gctacactag aaggttctcc aacggaagtc 240
agcggcaccc cattgtacgt tgggaagaac gtgttagaca attccaaagg cacaatggac 300
caagaacttc ttacgcctga gttcaactac acgtacactg aatctacgtc aaatactacg 360
acgcacgggt tgaagcttgg ggtgaaaacc actgcgacca tgaagtttcc tatagcacag 420
ggctcgatgg aggcctctac cgagtacaac ttccaggatt cctccacgga taccactaca 480
aaaacagtta gctacaagag tcctagtcag aagataaaag tccccgccgg aaaaacattt 540
cgggtccttg catatctcaa caccggtagc attagtggtg aagccaatct ctacgcaaat 600
gtcggtggaa tagcgtgggg cgttctcccc ggatatccaa acggtggagg cgtgaacatc 660
ggcgcggtgc tgacaaagtg ccagcaaaaa gggtggggcg attttcgcaa cttccagccg 720
tccggccgcg acgtgattgt gaagggacaa ggaacattca agtcgaacta tggtaccgat 780
ttcattttaa agatcgagga cattactgac agcaagctga gaaataacaa cggctctggg 840
actgttgttc aggagatcaa ggtaccattg attcgtacag agatctag 888
<210> 60
<211> 888
<212> DNA
<213> 人工序列-ACe1_21转基因植物核苷酸序列(Artificial Sequence)
<400> 60
atgtcatcta ccgatgtgca agaaaggctc cgcgacctcg cgagggagaa cgaggccggc 60
acgctcaatg aagcgtggaa cacaaatttc aagccttctg atgagcagca gttctcgtac 120
agcccaactg aaggaatagt gtttctgacg ccgccaaaaa atgtcatcgg cgagcggcgc 180
atatcacagt acaaagtcaa taatgcttgg gccacccttg aaggcagccc caccgaggtg 240
tcaggaactc ctctgtatgt aggcaagaat gttcttgaca actccaaggg taccatggac 300
caggagttgt tgacgcctga gttcaactac acctatacag aaagtacaag caatacaacg 360
acgcacggcc tcaaggtggg ggtgaagaca actgctacaa tgaagttccc catcgcgcaa 420
ggaagcatgg aggcatcaac agagtacaac ttccaggact cgtccactga tacaaccacc 480
aaaaccgtct cctacaagag cccgtcccag aagatcaaag ttccggcggg gaagactttt 540
cgtgtgctcg cctacctgaa cactggaagt atctctggag aagcaaacct atacgccaac 600
gttggcggga ttgcatgggg tgtactgccg gggtatccaa atggtggcgg cgtcaacatt 660
ggtgctgttt tatccaagtg ccagcaaaaa ggctggggcg actttagaaa ttttcagccc 720
agtggtcggg atgttattgt caagggacaa ggcaccttca aatctaacta tggtactgac 780
ttcatcctga agattgaaga tattacagat tcaaaactaa ggaacaacaa tggatcgggg 840
acggtggtgc aagagatcaa ggttcctctt ataagaactg agatatag 888

Claims (10)

1.一种控制亚洲玉米螟害虫的方法，其特征在于，包括将亚洲玉米螟害虫至少与ACe1蛋白接触；
优选地，所述ACe1蛋白存在于至少产生所述ACe1蛋白的宿主细胞中，所述亚洲玉米螟害虫通过摄食所述宿主细胞至少与所述ACe1蛋白接触；
更优选地，所述ACe1蛋白存在于至少产生所述ACe1蛋白的细菌或转基因植物中，所述亚洲玉米螟害虫通过摄食所述细菌或所述转基因植物的组织至少与所述ACe1蛋白接触，接触后所述亚洲玉米螟害虫生长受到抑制和/或导致死亡，以实现对亚洲玉米螟危害植物的控制。
2.根据权利要求1所述的控制亚洲玉米螟害虫的方法，其特征在于，所述转基因植物为玉米、高粱；
优选地，所述转基因植物的组织为根、叶片、茎秆、果实、雄穗、雌穗、花药或花丝。
3.根据权利要求1或2所述的控制亚洲玉米螟害虫的方法，其特征在于，所述ACe1蛋白为ACe1_3蛋白、ACe1_4蛋白、ACe1_5蛋白、ACe1_6蛋白、ACe1_8蛋白、ACe1_9蛋白、ACe1_10蛋白、ACe1_11蛋白、ACe1_12蛋白、ACe1_13蛋白、ACe1_14蛋白、ACe1_15蛋白、ACe1_16蛋白、ACe1_17蛋白、ACe1_18蛋白、ACe1_19蛋白、ACe1_20蛋白或ACe1_21蛋白；
优选地，所述ACe1蛋白氨基酸序列具有SEQ ID NO:1至SEQ ID NO:18所示的氨基酸序列；
更优选地，所述ACe1蛋白在细菌中的核苷酸序列具有SEQ ID NO:19至SEQ ID NO:36所示的核苷酸序列；所述ACe1蛋白在转基因植物中的核苷酸序列具有SEQ ID NO:43至SEQ IDNO:60所示的核苷酸序列。
4.根据权利要求1至3任一项所述的控制亚洲玉米螟害虫的方法，其特征在于，所述转基因植物还包括至少一种不同于编码所述ACe1蛋白的核苷酸的第二种核苷酸。
5.根据权利要求4所述的控制亚洲玉米螟害虫的方法，其特征在于，所述第二种核苷酸编码Cry类杀虫蛋白质、Vip类杀虫蛋白质、蛋白酶抑制剂、凝集素、α-淀粉酶或过氧化物酶；
优选地，所述第二种核苷酸编码Cry1Ab蛋白、Cry2Ab蛋白或Cry1A.105蛋白。
更优选地，所述Cry1Ab蛋白、Cry2Ab蛋白、Cry1A.105蛋白具有SEQ ID NO:37、SEQ IDNO:38、SEQ ID NO:39所示的氨基酸序列。
进一步优选地，所述第二种核苷酸具有SEQ ID NO:40、SEQ ID NO:41、SEQ ID NO:42所示的核苷酸序列。
6.根据权利要求4所述的控制亚洲玉米螟害虫的方法，其特征在于，所述第二种核苷酸为抑制目标昆虫害虫中重要基因的dsRNA。
7.一种ACe1蛋白质控制亚洲玉米螟害虫的用途。
8.一种产生控制亚洲玉米螟害虫的植物的方法，其特征在于，包括向所述植物的基因组中引入编码ACe1蛋白的多核苷酸序列。
9.一种产生控制亚洲玉米螟害虫的植物种子的方法，其特征在于，包括将由权利要求8所述方法获得的第一植株与第二植株杂交，从而产生含有编码ACe1蛋白的多核苷酸序列的种子。
10.一种培养控制亚洲玉米螟害虫的植物的方法，其特征在于，包括：
种植至少一粒植物种子，所述植物种子的基因组中包括编码ACe1蛋白的多核苷酸序列；
使所述植物种子长成植株；
使所述植株在人工接种亚洲玉米螟害虫和/或亚洲玉米螟害虫自然发生危害的条件下生长，收获与其他不具有编码ACe1蛋白的多核苷酸序列的植株相比具有减弱的植物损伤和/或具有增加的植物产量的植株。
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