CN115362370A - 新颖的杀昆虫毒素受体和使用方法 - Google Patents

Abstract

本公开涉及杀昆虫毒素抗性管理和筛选抗昆虫的新颖的杀昆虫毒素受体。一个实施例涉及与编码新颖的杀昆虫毒素受体的多核苷酸和由其编码的多肽有关的分离、表征、组合物和使用方法。多核苷酸和多肽可用于鉴定或设计昆虫有害生物管理策略。

Description

新颖的杀昆虫毒素受体和使用方法

以电子方式递交的序列表的引用

该序列表的官方副本经由EFS-Web作为ASCII格式的序列表以电子方式提交，文件名为“8428_SeqList.txt”，创建于2021年3月1日，具有291千字节大小，并且与本说明书同时提交。包含在所述ASCII格式的文件中的序列表是本说明书的一部分并且通过引用以其整体并入本文。

技术领域

本公开涉及杀昆虫毒素抗性管理和筛选抗昆虫的新颖的杀昆虫毒素受体。一个实施例涉及与编码新颖的杀昆虫毒素受体的多核苷酸和由其编码的多肽有关的分离、表征、组合物和使用方法。多核苷酸和多肽可用于鉴定或设计昆虫有害生物管理策略。

背景技术

昆虫有害生物是全世界农作物损失中的一个主要因素。例如，黏虫摄食、黑切根虫损害或欧洲玉米螟损害对农业生产者来说在经济上可能是破坏性的。仅来自在田间和甜玉米上的攻击的昆虫有害生物相关的作物损失每年在损失和控制费用方面已经达到约十亿美元。

传统上，种植者已经使用化学杀有害生物剂作为控制农业重要有害生物的手段。引入携带来自苏云金芽孢杆菌(Bacillus thuringiensis，Bt)的δ-内毒素的转基因植物提供了非化学的控制方法。

传统上，Bt毒素通过其对特定昆虫类别的特定毒性来分类。

鳞翅目昆虫对整个北美和世界的玉蜀黍作物造成相当大的损害。主要的有害生物之一是欧洲玉米螟(Ostrinia nubulalis)，通常称为欧洲玉米蛀虫(ECB)。编码来自Bt的晶体蛋白CrylAb和CrylAc的基因已经作为ECB控制和其他有害生物控制的手段被引入玉蜀黍中。这些转基因玉蜀黍杂合体已经有效地控制了ECB。然而，所形成的对Bt毒素的抗性对有害生物的控制提出了挑战。

鉴定Bt毒素的植物有害生物结合多肽可用于调研Bt毒素-Bt毒素受体相互作用、选择和设计改进的毒素或其他杀昆虫剂、开发新颖的杀昆虫剂、以及筛选抗性或其他抗性管理策略和工具。

发明内容

提供了用于调节昆虫对杀昆虫蛋白质的易感性的组合物和方法。这些组合物和方法涉及来自鞘翅目昆虫，例如西方玉米根虫(玉米根萤叶甲(Diabrotica virgiferavirgifera))的毒素受体多肽。

在此提供的方法可用于调研毒素受体的结构-功能关系；调研毒素-受体相互作用；阐明毒素的作用方式；筛选和鉴定包含新颖的杀昆虫毒素的新颖的毒素受体配体；设计并开发新颖的毒素受体配体；并产生对杀昆虫毒素具有改变的易感性的昆虫或昆虫群落。

本公开提供了用于检测对杀昆虫毒素具有抗性的昆虫的方法。在一些方面，这些方法涉及检测对Cry3类杀昆虫毒素具有抗性的西方玉米根虫(玉米根萤叶甲)。

本公开提供了用于检测作物田间对Cry3类杀昆虫蛋白质具有抗性的昆虫的方法，所述方法包括提供来自昆虫的样品；测定样品中存在的核酸；以及检测与SEQ ID NO：1相比改变的DvABCB1序列或降低的DvABCB1基因的转录物水平，其中该改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。在一些方面，描述的方法进一步包括在检测到Cry3抗性昆虫时改变作物田间的有害生物综合管理策略。在其他方面，描述的方法进一步包括在检测到Cry3抗性昆虫时向作物田间喷洒杀昆虫剂。在一些实施例中，改变的DvABCB1序列包含DvABCB1_3AR1(SEQ ID NO：3)、DvABCB1_3AR_2(SEQ ID NO：4)、或DvABCB1_3AR3(SEQ ID NO：5)、或者SEQ ID NO：40-63中的任一个。

本公开提供了用于检测作物田间对Cry3类杀昆虫蛋白质具有抗性的昆虫的方法，这些方法包括提供来自昆虫的样品；测定样品中存在的核酸；以及检测与SEQ ID NO：1相比具有缺失的改变的DvABCB1序列或降低的DvABCB1基因的转录物水平，其中该改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。在一些方面，描述的方法进一步包括在检测到Cry3抗性昆虫时改变作物田间的有害生物综合管理策略。在其他方面，描述的方法进一步包括在检测到Cry3抗性昆虫时向作物田间喷洒杀昆虫剂。

本公开提供了用于检测作物田间对Cry3类杀昆虫蛋白质具有抗性的昆虫的方法，这些方法包括提供来自昆虫的样品；测定样品中存在的核酸；以及检测与SEQ ID NO：1相比具有小于95％序列同一性的改变的DvABCB1序列或降低的DvABCB1基因的转录物水平，其中该改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。在一些方面，描述的方法进一步包括在检测到Cry3抗性昆虫时改变作物田间的有害生物综合管理策略。在其他方面，描述的方法进一步包括在检测到Cry3抗性昆虫时向作物田间喷洒杀昆虫剂。

本公开提供了用于检测作物田间对Cry3类杀昆虫蛋白质具有抗性的昆虫的方法，这些方法包括提供来自昆虫的样品；测定样品中存在的多肽；以及检测与SEQ ID NO：2相比具有小于95％序列同一性的改变的DvABCB1序列或降低的DvABCB1基因的蛋白质水平，其中该改变的DvABCB1序列或降低的DvABCB1的蛋白质水平显示Cry3抗性昆虫。在一些方面，描述的方法进一步包括在检测到Cry3抗性昆虫时改变作物田间的有害生物综合管理策略。在其他方面，描述的方法进一步包括在检测到Cry3抗性昆虫时向作物田间喷洒杀昆虫剂。

在一些方面，本公开涉及用于筛选昆虫群体的试剂盒，其中所述试剂盒包含用于检测与SEQ ID NO：1相比改变的DvABCB1序列或降低的DvABCB1基因的转录物水平的装置，其中该改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。

本文提供的组合物和方法也可用于管理植物有害生物中的毒素抗性，用于监测植物有害生物中的毒素抗性，以及用于保护植物免受植物有害生物的损害。

附图说明

图1：示出了来自Cry3A抗性WCR样品的DvABCB1_3AR1(SEQ ID NO：3)、DvABCB1_3AR_2(SEQ ID NO：4)、DvABCB1_3AR3(SEQ ID NO：5)和来自易感性WCR样品的DvABCB1_Sus1的DvABCB1 cDNA序列的比对。在DvABCB1_3AR1和DvABCB1_3AR2中分别鉴定出726bp和2430bp的大缺失，没有移码或终止密码子。由于24个SNP，DvABCB1_Sus1和DvABCB1_3AR3共享99.36％的同一性。

具体实施方式

如本文使用的，单数形式“一个/种(a/an)”以及“所述(the)”包括复数个指示物，除非上下文中另有明确指明。因此，例如，提及“细胞”包括多个此类细胞，并且提及“蛋白质”包括提及一种或多种蛋白及其等同物，等等。本文所用的所有技术和科学术语具有与本公开所属领域的普通技术人员通常所理解相同的含义，除非另有明确说明。

本文提供的实施例涉及具有毒素结合活性的受体多肽，这些受体衍生自玉米根萤叶甲并具有毒素结合活性。在一些实施例中，所述受体具有Bt结合活性。在其他实施例中，所述受体具有非Bt毒素结合活性。

因此，一个实施例提供了分离的核酸分子，其包含编码SEQ ID NO：1所示的具有毒素结合活性的多肽及其氨基酸取代、缺失、插入、片段及其组合的核苷酸序列。组合物包括具有毒素结合活性的多肽的核酸分子编码序列、包含那些核酸分子的载体、以及包含这些载体的宿主细胞。进一步提供了本文所述的片段和变体多肽。

一个实施例涵盖分离的或实质纯化的核酸或多肽组合物。“分离的”或“纯化的”核酸分子或多肽、或其生物活性部分实质上不含其他细胞物质，或者当通过重组技术产生时实质上不含培养基，或者当经化学合成时实质上不含化学前体或其他化学品。“分离的”核酸可以不含在衍生出所述核酸的生物体的基因组DNA中天然地位于所述核酸侧翼的序列(即，位于所述核酸的5′和3′末端的序列)(优选编码多肽的序列)。例如，在一个实施例中，分离的核酸分子可以包含小于约5kb、4kb、3kb、2kb、1kb、0.5kb、或0.1kb的核酸序列，该核酸序列在衍生出该核酸的细胞的基因组DNA中天然地位于该核酸分子的侧翼。一个实施例考虑实质上不含细胞物质的多肽，该细胞物质包括具有小于约30％、20％、10％、5％(以干重计)的污染多肽的多肽制剂。当重组产生多肽或其生物活性部分时，培养基可以代表小于约30％、20％、10％、或5％(以干重计)的化学前体或非目的多肽的化学品。

在另一个实施例中，多肽制剂可以包含不干扰多肽具体所需活性的污染物质。组合物还涵盖所公开的核苷酸序列的片段和变体以及由此编码的多肽。

除了其他用途之外，多核苷酸组合物可用于在目的细胞中表达受体多肽或其片段，以产生所述多肽的细胞或分离制剂，用于调研毒素受体的结构-功能和/或序列-功能关系，评估毒素-受体相互作用，阐明毒素的作用方式，筛选测试化合物以鉴定包括新颖的杀昆虫毒素的新颖的毒素受体配体，并且设计并开发包括新颖的杀昆虫毒素的新颖的毒素受体配体。

编码实施例的受体多肽的分离的多核苷酸可以在目的细胞中表达；并且所产生的毒素受体多肽可用于完整的细胞或体外受体结合测定、和/或完整细胞毒性测定。

如本文所用，“Bt毒素”是指编码苏云金芽孢杆菌蛋白质、其衍生物或基于其建模的合成多肽的基因。参见，例如，Geiser等人，(1986)Gene[基因]48：109，其公开了Bt δ-内毒素基因的克隆和核苷酸序列。此外，编码δ-内毒素基因的DNA分子可购自美国典型培养物保藏中心(American Type Culture Collection)(美国马里兰州罗克韦尔市(Rockville，Md.))，例如

登录号40098、67136、31995和31998下。这些类别的苏云金芽孢杆菌杀昆虫蛋白的成员包括但不限于：本领域技术人员熟知的Cry蛋白(参见，Crickmore，等人，“Bacillus thuringiensis toxin nomenclature[苏云金芽孢杆菌毒素命名法]”(2011)，在可以使用“www”前缀在万维网上访问的btnomenclature.info)。

“目的细胞”是指其中本文公开的多肽的表达是所希望的任何细胞。目的细胞包括但不限于：哺乳动物、禽类、昆虫、植物、细菌、真菌和酵母细胞。目的细胞包括但不限于：所培养的细胞系，原代细胞培养物、体内细胞、和转基因或遗传修饰的或编辑的生物体的细胞。

如本文所用，“修饰的”或“改变的”序列是指与野生型序列不同的序列。在一个实施例中，修饰或改变的多核苷酸序列与SEQ ID NO：1不同。在另一个实施例中，修饰或改变的氨基酸序列与SEQ ID NO：2或SEQ ID NO：40-63中的任一个不同。在一个实施例中，可以筛选序列中的修饰或改变以确定对毒素的改变的易感性。所实施的方法考虑了本文公开的多肽和多核苷酸在受体结合和/或毒性测定中筛选测试化合物以鉴定新颖的毒素受体配体(包括受体激动剂和拮抗剂)或筛选抗性的用途。测试化合物可以包括可从由组合合成方法产生的小分子的不同文库获得的分子。测试化合物还可以包括但不限于：抗体、结合肽、和被设计或推断与本实施例的受体多肽相互作用的其他小分子。测试化合物还可以包括受体的肽片段、抗受体抗体、模仿毒素的一个或多个受体结合结构域的抗独特型抗体、结合肽、嵌合肽、和融合物、或异源多肽，其通过将两个或更多个毒素或其片段，如本文公开的受体的细胞外部分等组合而产生。通过实施例的筛选方法所鉴定的配体包括潜在的新颖的杀昆虫毒素，其杀昆虫活性可以确定(参见例如，美国专利号5,407,454、5,986,177和6,232,439)。

在一个实施例中，方法涉及分离昆虫中肠毒素的受体，这些方法包括解剖昆虫中肠组织；对该昆虫中肠组织进行膜富集步骤，如BBMV制备；用昆虫毒素在富集的膜上进行溶液内结合测定；并且进行亲和纯化，其中该毒素是该亲和纯化靶标。在另一个实施例中，可以对整个昆虫进行膜富集步骤。在另一个实施例中，亲和纯化可以在溶液内结合步骤之前进行。在一个实施例中，亲和纯化靶标是昆虫毒素。在另一个实施例中，亲和纯化靶标是受体多肽。

该实施例提供了用于筛选与本文公开的多肽结合的配体的方法。多肽及其片段(例如，毒素结合肽)都可以用于化合物的筛选测定，这些化合物与受体肽结合并且表现出所需结合特征。所需结合特征包括但不限于：结合亲和力、结合位点特异性、缔合和解离速率等。筛选测定可以在完整细胞中或在体外测定中进行，这些测定包括将配体结合结构域暴露于样品配体并检测配体结合多肽复合物的形成。测定可以是直接配体-受体结合测定，配体竞争测定、或被设计用于测量结合对转运蛋白功能(例如ATP水解，构象变化、或溶质转运)的影响的间接测定。在一些实施例中，修饰的或编辑的受体可以用于化合物的筛选测定，这些化合物与修饰的或编辑的受体肽结合并且表现出所需结合特征。

这些方法包括提供至少一个本文公开的毒素受体多肽，在促进结合的条件下，将多肽与样品和对照配体接触，并确定样品配体相对于对照配体的结合特征。针对体外结合测定，多肽可以作为分离的、裂解的、或均质的细胞制剂来提供。分离的多肽可以在溶液中提供，或固定在基质上。用于固定多肽的方法包括但不限于具有市售高亲和力配体的融合多肽的构建和使用。例如，GST融合蛋白可以吸附在谷胱甘肽琼脂糖凝胶珠(西格玛化学品公司(Sigma Chemical)，圣路易斯，密苏里州)上或谷胱甘肽衍生的微量滴定板上。也可以如下固定多肽：使用生物素和链霉亲和素，将多肽化学缀合(连接)到基质，或通过使用多肽的抗体。可替代地，这些多肽可以在完整的细胞结合测定中提供，其中这些多肽通常表达为细胞表面毒素受体。

本公开提供了使用完整细胞毒性测定来筛选与本文公开的受体多肽结合的配体并对在杀昆虫毒素的存在下表达多肽的目的细胞赋予毒性的方法。通过该筛选所选择的配体是对于表达受体多肽、修饰的或编辑的受体多肽、或其片段的昆虫的潜在的杀昆虫毒素。具体毒素的昆虫特异性可以通过特定昆虫物种中受体的存在来确定。毒素的结合可能对一些昆虫物种的受体是特异性的，并且同时对其他变体受体是不显著的或非特异性的。毒性测定包括在表达实施例的多肽的完整细胞中将多肽的毒素结合结构域暴露于样品配体并检测在表达多肽的细胞中受影响的毒性。“毒性”是指降低的细胞活力。“活力”是指在不存在特定细胞毒剂的情况下细胞以该细胞特征的方式增殖和/或分化和/或维持其生物特征的能力。

在一个实施例中，这些方法包括提供至少一种包含SEQ ID NO：2或40-63中的任一个的细胞表面毒素受体多肽或其细胞外毒素结合结构域，在促进结合的条件下将受体多肽与样品和对照配体接触，并相对于对照配体，确定表达细胞表面毒素受体多肽的细胞的活力。“接触”是指将样品和对照试剂呈递到实施例的多肽的预期配体结合位点。“促进结合的条件”是指物理和生物化学条件的任何组合，其使本实施例的多肽的配体能够在背景水平上结合预期多肽。在此方面，也可以使用用于研究蛋白质-蛋白质相互作用的市售方法，如酵母和/或细菌双杂交系统。双杂交系统可从例如Clontech(帕洛阿尔托，加利福尼亚州)或Display Systems Biotech公司(维斯塔(Vista)，加利福尼亚州)获得。

本文公开的组合物和筛选方法可用于设计和开发包括新颖的杀昆虫毒素的新颖的毒素受体配体。各种候选配体；针对结合、毒性、和物种特异性所筛选和表征的配体；和/或具有已知特征和特异性的配体可以被连接或修饰以产生具有特别期望的特征和特异性的新颖的配体。用于评估结合、毒性和杀昆虫活性的本文所述的方法可以用于筛选和表征新颖的配体。

本文公开的组合物和筛选方法可用于设计和开发新颖的毒素受体-配体复合物，其中受体和配体两者都在同一细胞中表达。“复合物”是指受体与配体的缔合足以防止与细胞中配体的其他相互作用。受体可以是本文所述的受体或其变体或片段。此外，受体可以是保留本文所述的受体多肽的生物学活性的异源多肽。

在一个实施例中，将编码受体的序列及其变体和片段与酵母和细菌双杂交系统一起使用以筛选目的毒素(例如，更具体和/或更有效的毒素)或筛选结合受体并可用于开发新颖的杀昆虫剂的昆虫分子。

“连接”是指在两个或更多个分子之间产生的共价键。可用于修饰和/或连接多肽配体如毒素的方法包括诱变和引起重组方法，包括但不限于定点诱变、嵌合多肽构建、和DNA改组。多肽修饰方法还包括用于共价修饰多肽的方法。如本文使用的，术语“可操作地连接”是指例如启动子和第二序列之间的功能性连接，其中启动子序列启动并介导相应于第二序列的DNA序列的转录。通常，可操作地连接意味着所连接的核酸序列是连续的，并且在必要时在相同阅读框中连接两个蛋白质编码区域。

所述组合物和筛选方法可用于将配体靶向表达受体或修饰的受体多肽的细胞。为了靶向，将不结合受体多肽的次级多肽和/或小分子与结合本文公开的受体多肽的一种或多种主要配体(包括但不限于Cry3毒素)连接。通过连接，可以将连接到主要配体的任何多肽和/或小分子靶向受体多肽，从而靶向表达受体多肽的细胞；其中配体结合位点可在细胞的细胞外表面获得。

为了在目的细胞中表达SEQ ID NO：2的毒素受体多肽、变体或片段，这些毒素受体序列可以在表达盒中提供。所述盒可以包括可操作地连接到毒素受体序列上的5’和3’调节序列。所述盒可以另外含有至少一个待共转化到生物体中的另外的基因。可替代地，一个或多个另外的基因可以在多个表达盒上提供。

这样的表达盒可具有多个限制位点，用于使所述毒素受体序列的插入受调节区的转录调节。表达盒可另外含有选择性标记基因。

所述表达盒可以5′-3’转录方向包含转录和翻译起始区域(即启动子)、毒素受体核苷酸序列、和在宿主细胞中起作用的转录和翻译终止区域(即终止区域)。转录起始区域，启动子，对植物宿主和/或对毒素受体序列可以是天然的或类似的，或者是外源的或异源的。此外，所述启动子可以是天然序列，或可替代地，是合成序列。当序列是“外源的”或“异源的”时，意指所述序列不是天然的或天然存在的。如本文使用的，嵌合基因包含与转录起始区可操作地连接的编码序列，所述转录起始区对于所述编码序列是异源的。如本文所用，修饰或编辑的序列是异源序列。

异源启动子或天然启动子序列可用于构建体设计。此类构建体可以改变目的细胞中毒素受体的表达水平，导致细胞表型的改变。

终止区对于转录起始区可以是天然的，对于可操作地连接的目的DNA序列可以是天然的，对于植物宿主可以是天然的，或者可以源自另一种来源(即，对于启动子、目的毒素受体序列、植物宿主、或其任何组合而言是外源的或异源的)。方便的终止区可获得自根癌农杆菌(A.tumefaciens)的Ti质粒，如章鱼碱合酶和胭脂碱合酶终止区。还参见Guerineau等人(1991)Mol.Gen.Genet.[分子遗传学和普通遗传学]262：141-144；Proudfoot(1991)Cell[细胞]64：671-674；Sanfacon等人，(1991)Genes Dev.[基因与发育]5：141-149；Mogen等人(1990)Plant Cell[植物细胞]2：1261-1272；Munroe等人，(1990)Gene[基因]，91：151-158；Ballas等人，(1989)Nucleic Acids Res.[核酸研究]17：7891-7903；以及Joshi等人(1987)Nucleic Acids Res.[核酸研究]15：9627-9639。

在适当的情况下，可以优化基因以在特定的目的转化细胞中增加表达。也就是说，可以使用宿主细胞优选的密码子来合成基因以改进表达。

另外的序列修饰可增强细胞宿主中的基因表达。这些包括消除以下序列：编码假聚腺苷酸化信号、外显子-内含子剪接位点信号、转座子样重复的序列、及可能不利于基因表达的其他经充分表征的序列。可以将序列的G-C含量调整至通过参照宿主细胞中表达的已知基因而计算出的给定细胞宿主的平均水平。如果可能，修饰序列以避免出现预测的发夹二级mRNA结构。

表达盒可另外在表达盒构建体中包含5′前导序列。此类前导序列可以起到增强翻译的作用。翻译前导序列包括：小核糖核酸病毒前导序列，例如EMCV前导序列(脑心肌炎5’非编码区；Elroy-Stein等人(1989)PNAS USA[美国科学院院报]86：6126-6130)；马铃薯Y病毒组前导序列，例如，TEV前导序列(烟草蚀纹病毒；Allison等人(1986))；MDMV前导序列(玉蜀黍矮小花叶病毒)、及人类免疫球蛋白重链结合多肽(BiP)(Macejak等人(1991)Nature[自然]353：90-94)；来自苜蓿花叶病病毒的外壳多肽mRNA的非翻译前导序列(AMV RNA 4)(Jobling等人(1987)Nature[自然]325：622-625)；烟草花叶病毒前导序列(TMV；Gallie等人(1989)在Molecular Biology of RNA[RNA的分子生物学]中，编辑Cech(利斯公司(Liss)，纽约)，第237-256页)；和玉蜀黍褪绿斑驳病毒前导序列(MCMV；Lommel等人(1991)Virology[病毒学]81：382-385)。还参见，Della-Cioppa等人(1987)Plant Physiol.[植物生理学]84：965-968。还可以利用其他增强翻译的方法，例如内含子等。

在制备表达盒时，可以操作各种DNA片段，以提供处于适当取向以及合适时，处于适当阅读框中的DNA序列。为此，可采用衔接子(adapter)或接头以连接DNA片段，或可以涉及其他操作以提供方便的限制位点、移除多余的DNA、移除限制位点等。出于这个目的，可以涉及体外诱变、引物修复、限制性酶切(restriction)、退火、再取代(例如转换和颠换)。

使用本文公开的核酸，多肽可以在任何目的细胞中表达，细胞的具体选择取决于如下因素，如所需表达水平和/或受体活性。目的细胞包括但不限于：哺乳动物、植物、昆虫、细菌、和酵母宿主细胞。启动子、终止子、增强子和其他表达载体组分的选择也将取决于所选择的细胞。细胞在非天然条件下(例如数量，组成，位置和/或时间)产生蛋白质，因为它们已通过人为干预进行遗传改变而这样做。

选择载体以允许导入适当的宿主细胞中。细菌载体典型地为质粒或噬菌体来源的。用噬菌体载体颗粒感染或用裸的噬菌体载体DNA转染适当的细菌细胞。如果使用质粒载体，则用质粒载体DNA转染细菌细胞。用于表达本实施例的蛋白质的表达系统可以使用芽孢杆菌属物种和沙门氏菌属。参见，Palva等人(1983)Gene[基因]22：229-235和Mosbach等人(1983)Nature[自然]302：543-545。

各种真核表达系统如酵母、昆虫细胞系、植物和哺乳动物细胞可用于表达。本文公开的序列可以在这些真核系统中表达。在一些实施例中，采用转化/转染的植物细胞作为用于生产受体蛋白质的表达系统。

本文公开的编码多肽的序列也可以连接到用于转染例如哺乳动物、昆虫、或植物来源的细胞培养物的各种表达载体。用于生产肽的细胞培养物的示例是哺乳动物细胞。哺乳动物细胞系统通常将是单层细胞的形式，尽管也可使用哺乳动物细胞悬浮液。本领域已经开发了许多能够表达完整蛋白质的合适的宿主细胞系，并且包括COS、HEK293、BHK21、和CHO细胞系。这些细胞的表达载体可以包括表达控制序列，如复制起点、启动子(例如，CMV启动子，HSV tk启动子或pgk(磷酸甘油酸酯激酶启动子))、增强子(Queen等人(1986)Immunol.Rev.[免疫学研究]89：49)，以及必需的加工信息位点，如核糖体结合位点、RNA剪接位点、聚腺苷酸化位点(例如，SV40大T Ag聚A添加位点)和转录终止子序列。可获得可用于生产蛋白质的其他动物细胞，例如，自美国典型微生物保藏中心细胞系和杂交瘤目录(第7版，1992)。用于表达毒素受体并评估由所述受体介导的毒素细胞毒性的哺乳动物细胞的一个实例是人胚胎肾293细胞。参见美国专利号5,693,491，其通过引用并入本文。

用于在昆虫细胞中表达本文公开的多肽的合适载体通常衍生自SF9杆状病毒。合适的昆虫细胞系包括蚊幼虫、蚕、黏虫、蛾和果蝇细胞系如Schneider细胞系(Schneider等人(1987)J.Embryol.Exp.Morphol.[胚胎学与实验学杂志]27：353-365)。一个实施例考虑了无细胞多肽表达系统。

与酵母一样，当采用高等动物或植物宿主细胞时，通常将聚腺苷酸化或转录终止子序列并入载体中。终止子序列的实例是来自牛生长激素基因的聚腺苷酸化序列。还可以包括用于准确剪接转录物的序列。剪接序列的实例是来自SV40的VP1内含子(Sprague等人(1983)J.Virol.[病毒学杂志]45：773-781)。此外，可将控制宿主细胞中的复制的基因序列引入到载体中，如在牛乳头瘤病毒型载体中发现的那些。Saveria-Campo，M.，BovinePapilloma Virus DNA a Eukaryotic Cloning Vector in DNA Cloning Vol.II aPractical Approach[牛乳头瘤病毒DNA：DNA克隆中的真核克隆载体，第II卷，实用方法]，D.M.Glover编辑，IRL Pres[IRL出版社]，美国弗吉尼亚州阿林顿，第213页至第238页(1985)。

在一个具体实施例中，可能需要负面控制受体结合；特别是当不再需要对细胞的毒性或者希望将毒性降低到较低水平时。在这种情况下，配体-受体多肽结合测定可用于筛选结合受体多肽但不对表达受体的细胞赋予毒性的化合物。可用于阻断配体结合的分子的实例包括一种抗体，该抗体特异性识别受体多肽的配体结合结构域，这样使得根据需要减少或阻止配体结合。

在另一个实施例中，可以改变受体多核苷酸或多肽表达，例如通过介导RNA干扰(RNAi)的还原，包括使用针对特异性受体多核苷酸序列的沉默元件。沉默元件可以包括但不限于：正义抑制元件、反义抑制元件、双链RNA(dsRNA)、siRNA、amiRNA、miRNA、或发夹抑制元件。通过沉默元件抑制受体多核苷酸或多肽的编码序列的表达可以通过向昆虫提供外源核酸沉默元件构建体，例如dsRNA来进行。沉默元件构建体包含靶向受体多核苷酸的至少一个沉默元件。

在具体实施例中，减少有害生物中靶序列的多核苷酸水平和/或多肽水平导致在适当的靶昆虫中的小于95％、小于90％、小于80％、小于70％、小于60％、小于50％、小于40％、小于30％、小于20％、小于10％、或小于5％的相同靶序列的多核苷酸水平，或由其编码的多肽的水平。测定RNA转录物水平的方法包括但不限于：qRT-PCR、RNA印迹法、RT-PCR、和数字PCR。

在具体实施例中，沉默元件与靶受体多核苷酸具有100％序列同一性。在其他实施例中，沉默元件与靶多肽具有同源性，该靶多肽与靶多核苷酸的区域具有至少50％、60％、70％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％、或更大序列同一性，其中与靶多核苷酸的序列同一性仅需足以降低靶受体多核苷酸的表达即可。通常，可以使用至少19个核苷酸、21个核苷酸、24个核苷酸、50个核苷酸、100个核苷酸、200个核苷酸、或更多核苷酸的序列。

本文考虑了所公开的核苷酸序列和由其编码的多肽的片段和变体。“片段”是指核苷酸序列的一部分，或氨基酸序列的一部分，以及因此由此编码的多肽的一部分。核苷酸序列的片段可以编码多肽片段，这些多肽片段保留天然多肽的生物活性并且例如结合毒素，包括Bt毒素。可替代地，核苷酸序列的片段可用作杂交探针。因此，核苷酸序列的片段范围可以是从至少约20个核苷酸、约50个核苷酸、约100个核苷酸、至多达与编码本实施例的多肽的全长核苷酸序列相比少一个核苷酸。

编码毒素受体多肽的生物活性部分的毒素受体核苷酸序列的片段可以编码至少15、25、30、50、100、150、200或250个连续氨基酸，或多达与全长毒素受体多肽中存在的氨基酸的总数相比少一个氨基酸的连续氨基酸。用作PCR引物的杂交探针的毒素受体核苷酸序列的片段通常不需要编码毒素受体多肽的生物活性部分。

因此，毒素受体核苷酸序列的片段可以编码毒素受体多肽的生物活性部分，或者它可以是可以使用下文公开的方法用作杂交探针或PCR引物的片段。毒素受体多肽的生物活性部分可以通过如下来制备：分离毒素受体核苷酸序列之一的一部分，表达毒素受体多肽的编码部分(例如，通过体外重组表达)，并且评估毒素受体多肽的编码部分的活性。作为毒素受体核苷酸序列的片段的核酸分子包含至少16、20、50、75、100、150、200、250、300、350、400、450、500、550、600、650、700、800、900、1,000、1,100、1,200、1,300、1,400、1500、2000、或2500个核苷酸，或多达与本文公开的全长毒素受体核苷酸序列中存在的核苷酸的数目相比少一个核苷酸。

“变体”是指实质相似的序列。对于核苷酸序列，由于遗传密码的简并性，保守变体包括编码毒素受体多肽之一的氨基酸序列的那些序列。可以使用熟知的分子生物学技术来鉴定天然存在的等位基因变体如，例如聚合酶链反应(PCR)和杂交技术鉴定。变体核苷酸序列还包括合成地衍生的核苷酸序列，如，例如通过利用定点诱变产生的，但是仍然编码毒素受体蛋白质的那些。通常，实施例的特定核苷酸序列的变体将与特定核苷酸序列具有至少约40％、50％、60％、65％、70％，通常至少约75％、80％、85％、86％、87％、88、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％、或99％或更高的序列同一性，如通过本文别处描述的序列比对程序使用默认参数所确定的。

还可以通过比较由变体核苷酸序列编码的多肽与由参比核苷酸序列编码的多肽之间的百分比序列同一性来评估实施例的特定核苷酸序列(即，参比核苷酸序列)的变体。因此，例如，公开了编码与SEQ ID NO：1、3、5或40-63中的任一个所示的多肽具有给定百分比序列同一性的多肽的分离的核酸。可以使用默认参数使用本文别处描述的序列对齐程序来计算任何两个多肽之间的序列同一性百分比。在本文公开的任何给定的多核苷酸对通过比较其编码的两个多肽共有的百分比序列同一性来评估的情况下，编码的两个多肽之间的百分比序列同一性为至少约40％、45％、50％、55％、60％、65％、70％，通常至少约75％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更高的序列同一性。在一些实施例中，提供了编码嵌合多肽的变体多核苷酸，所述嵌合多肽包含与本公开的第二毒素受体多肽的C-末端区域可操作地融合的本公开的第一毒素受体多肽的N-末端区域。

还可以通过比较由变体核苷酸序列编码的多肽与由参比核苷酸序列编码的多肽之间的序列同一性百分比来评估本文公开的特定核苷酸序列(即，参比核苷酸序列)的变体。因此，例如，公开了编码与SEQ ID NO：1、3、5、或40-63的多肽具有给定百分比序列同一性的多肽的分离的核酸。可以使用默认参数使用本文别处描述的序列对齐程序来计算任何两个多肽之间的序列同一性百分比。在任何给定的多核苷酸对通过比较其编码的两个多肽共有的百分比序列同一性来评估的情况下，编码的两个多肽之间的百分比序列同一性为至少约40％、45％、50％、55％、60％、65％、70％，75％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更高的序列同一性。

“变体”蛋白质是指通过以下方式衍生自天然蛋白质的蛋白质：在天然蛋白质的N-末端和/或C-末端缺失(所谓的截短)一个或多个氨基酸或添加一个或多个氨基酸；在天然蛋白质中的一个或多个位点处缺失或添加一个或多个氨基酸；或在天然蛋白质中的一个或多个位点处取代一个或多个氨基酸。本实施例中的变体多肽和多核苷酸还包括同源和直向同源多肽序列。本文考虑的变体蛋白是生物活性的，即它们仍然具有天然蛋白质的所需生物学活性，即本文所述的活性(例如，Bt毒素结合活性)。此类变体可以由例如遗传多态性或人为操作产生。天然毒素受体蛋白的生物活性变体将与天然蛋白的氨基酸序列具有至少约40％、50％、60％、65％、70％，通常至少约75％、80％、85％、86％、87％、88％、89％，如至少约90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更高的序列同一性，如本文别处所描述的序列比对程序使用默认参数所确定的。蛋白质的生物活性变体与该蛋白质可能相差仅仅1-15个氨基酸残基，仅仅1-10个，如6-10个，仅仅5个，仅仅4、3、2个，或甚至1个氨基酸残基。

在一个实施例中，靶受体的变体可用于筛选和/或检测，如但不限于噬菌体展示、PCR、RT-PCR、qRT-PCR、ELISA、Bradford测定、DNA印迹法、蛋白质印迹或RNA印迹法等。此筛选和/或检测可用于发展毒性增加的杀昆虫剂，筛选新颖的作用位点，或检测对杀昆虫蛋白质具有抗性的昆虫。筛选和/或检测也可用于筛选和/或检测昆虫或昆虫群体对杀昆虫剂的改变的易感性、改变的受体序列或改变的受体基因表达(转录物或蛋白质)。此外，多于一个变体，片段，受体或变体、片段或受体的组合可用于一个大的但多重的筛选测定中。

可以以各种方式改变实施例的多肽，这些方式包括氨基酸取代、缺失、截短、和插入。用于此类操作的方法是本领域通常已知的。例如，可以通过DNA中的突变来制备毒素受体多肽的氨基酸序列变体。可以进行保守取代，如将一个氨基酸与具有相似性质的另一个氨基酸交换。

因此，本文考虑的多肽和核苷酸序列包括天然存在的序列以及突变形式。同样，本实施例的蛋白质涵盖天然存在的蛋白质以及其变体和修饰的或编辑的形式。这些变体可以继续具有所需的毒素结合活性。

本文涵盖的蛋白质序列的缺失、插入、和取代预期不会产生蛋白质特征的根本性变化。

例如，认识到至少约10、20、50、100、150、200、250、300、350、400、450、500、550、600、650、700、750、800、850、900、950、和多达960个氨基酸可以从具有SEQ ID NO：2中所示氨基酸序列的多肽的N-末端缺失，并且仍保留结合功能。进一步认识到，至少约10、20、30、40、50、60、70、80、90、100、110、和多达119个氨基酸可以从具有SEQ ID NO：2中所示氨基酸序列的多肽的C-末端缺失，并且仍保留结合功能。缺失变体涵盖具有这些缺失的多肽。认识到保留结合功能的缺失变体涵盖具有这些N-末端或C-末端缺失，或在C-和N-末端具有其任何缺失组合的多肽。在一个实施例中，蛋白质序列的缺失、插入、和/或取代可以改变或表示对毒素的易感性的改变。可以通过受体结合和/或毒性测定来评估活性。

变体核苷酸序列和多肽还涵盖衍生自诱变和引起重组过程如DNA改组的序列和多肽。用这样的程序，可以操纵一种或多种不同的毒素受体编码序列以产生新的毒素受体，这些新的毒素受体包括但不限于：具有所需的性质的新的毒素受体。用这种方式，从相关序列多核苷酸的群体产生重组多核苷酸文库，这些相关序列多核苷酸包括如下序列区域，这些序列区域具有实质的序列同一性并且可以在体外或体内进行同源重组。例如，使用此方法，编码目的结构域的序列基序可以在毒素受体基因和其他已知的毒素受体基因之间进行改组，以获得编码具有改善的目的性质(如在受体的情况下增加的或降低的配体亲和力)的多肽的新基因。

当受体多肽在细胞中表达并与细胞膜缔合(例如，通过跨膜区段)时，为了使受体结合所需配体，例如Cry3毒素，受体的配体结合结构域对于配体而言必须是可获得的。在此方面，认识到将天然毒素受体定向，这样使得毒素结合位点可以在细胞外获得。

因此，在包括使用完整细胞的方法中，该实施例提供了细胞表面毒素受体。“细胞表面毒素受体”是指包含至少一个细胞外毒素结合位点的膜结合受体多肽。该实施例的细胞表面受体包括信号序列和跨膜区段的适当组合，用于在细胞膜处引导和保留受体，这样使得毒素结合位点可在细胞外获得。当天然的毒素受体用于表达，推断信号序列和跨膜区段的组成和构型时，没有必要确保多肽在细胞外显示毒素结合位点的适当拓扑结构。作为天然信号和跨膜序列的替代方案，异源信号和跨膜序列可用于产生细胞表面受体多肽。

认识到，可能感兴趣的可以是产生在细胞质中、在细胞核或其他胞器中、在其他亚细胞空间中；或在细胞外空间中能够与受体的配体发生细胞内相互作用的毒素受体。因此，该实施例涵盖受体的变体，其中受体多肽的一个或多个区段被修饰以将多肽靶向所需的细胞内或细胞外位置。

除其他事项之外，还涵盖用作与本文公开的细胞表面受体竞争的结合拮抗剂的受体片段和变体。例如，这样的片段或变体可以，例如，结合毒素但不能为特定细胞赋予毒性。在此方面，实施例提供了分泌的毒素受体，即，不与膜结合的受体。在另一个实施例中，除其他事项之外，受体片段和变体可用作与毒素具有协同关系的结合拮抗剂。分泌型受体可以包含异源或同源的信号序列，促进其从表达受体的细胞分泌；并且进一步包括对应于跨膜区段的区域的分泌变化。“分泌变化”是指对应于膜结合受体的跨膜区段的氨基酸包含一个或多个缺失、取代、插入、或其任何组合；这样使得该区域不再保留必要的疏水性以用作跨膜区段。可以通过确认包括来自表达多肽的细胞的变化的多肽的分泌来测试产生分泌变化的序列改变。

实施例的多肽可以从天然表达它们的细胞纯化，从被改变以表达它们的细胞中纯化(例如，重组宿主细胞)或使用多肽合成技术合成。在一个实施例中，通过重组DNA方法产生多肽。在此类方法中，将编码该多肽的核酸分子克隆到本文更全面描述的表达载体中，并且在适当的宿主细胞中表达。然后使用多肽纯化技术从细胞中分离多肽。可替代地，可以使用肽合成方法合成多肽或片段。

本文还考虑了其中一种或多种多肽与至少一种目的多肽融合的异源多肽。一个实施例涵盖融合多肽，其中目的异源多肽具有与受体多肽实质上不同源的氨基酸序列。在该实施例中，受体多肽和目的多肽可以或不可以可操作地连接。可操作连接的一个实例是框架内融合，这样使得在翻译时产生单个多肽。这样的融合多肽可以例如促进重组多肽的纯化。

在另一个实施例中，目的融合多肽在N末端可以包含异源信号序列，有利于其从特异性宿主细胞的分泌。因此可以通过使用异源信号序列增加多肽的表达和分泌。

该实施例还涉及多肽，其中本文所述多肽中的一个或多个结构域可操作地连接到具有同源功能的异源结构域。因此，毒素结合结构域可以用其他毒素的毒素结合结构域代替。因此，受体的毒素特异性基于除由毒素受体编码的结构域之外的毒素结合结构域，但是多肽的其他特征、例如膜定位和拓扑结构基于SEQ ID NO：2的毒素受体。

可替代地，可以保留天然毒素结合结构域，而受体包含另外的异源配体结合结构域，包括但不限于：异源毒素结合结构域。因此，本文还考虑了融合多肽，其中目的多肽是包括异源毒素结合结构域的异源多肽。包含Cry1毒素结合结构域的异源多肽的实例包括但不限于以下各项中所公开的那些：Knight等人(1994)Mol.Micro.[分子微生物学]11：429-436；Lee等人(1996)Appl.Environ.Micro.[应用与环境微生物学]63：2845-2849；Gill等人(1995)J.Biol.Chem.[生物化学杂志]270：27277-27282；Garczynski等人(1991)Appl.Environ.Microbiol.[应用与环境微生物学]10：2816-2820；Vadlamudi等人(1995)J.Biol.Chem.[生物化学杂志]270(10)：5490-4，和美国专利号5,693,491。

本文考虑的多肽变体包括包含增强或减少一个或多个结构域功能的突变的那些。例如，在毒素结合结构域中，可以引入增加或降低结构域对特定毒素的敏感性的突变。

作为引入突变的替代方案，可以通过增加配体结合结构域的拷贝数来实现活性的增加。因此，该实施例还涵盖受体多肽，其中以多于一个拷贝提供毒素结合结构域。

该实施例进一步涵盖包含受体表达载体的细胞，这些受体表达载体包含毒素受体序列、及其片段和变体。表达载体可以包含用于转化目的细胞的一个或多个表达盒。这些基因的转录可以置于组成型或诱导型启动子的控制下(例如，组织或细胞周期优选的)。

当使用多于一种的表达盒时，除包括至少一个受体序列的盒外的盒可以包括本文公开的受体序列或任何其他所需序列。

本文公开的核苷酸序列可用于分离昆虫物种(除玉米穗虫、大豆尺蠖、草地贪夜蛾、以及欧洲玉米螟以外，具体而言是其他鳞翅目物种，更具体而言是其他夜蛾科或草螟科物种)中的同源序列。

相对于参考序列(主题序列)，“序列同一性百分比(％)”被确定为在比对序列并引入空位(如果需要)以实现最大百分比序列同一性后，并且不考虑作为序列同一性的一部分的任何氨基酸保守取代，候选序列(查询序列)中与参考序列中的相应氨基酸残基或核苷酸相同的氨基酸残基或核苷酸的百分比。用于确定序列同一性百分比目的而进行的比对能以本领域技术范围内的各种方式实现，例如，使用公共可用的计算机软件，例如BLAST、BLAST-2。本领域的技术人员可以确定用于比对序列的适当参数，包括在进行比较的序列的全长度上实现最大比对所需的任何算法。两个序列之间的同一性百分比是序列共有的相同位置的数目的函数(例如，查询序列的同一性百分比＝查询序列和主题序列之间的相同位置的数目/查询序列的位置总数×100)。

本文公开的核苷酸序列可用于从其他生物体，特别是其他昆虫，更特别是其他鳞翅目或鞘翅目物种分离相应的序列。以这种方式，可以使用如PCR、杂交等方法基于该序列与本文所述序列的序列同源性来鉴定此类序列。此外，可以使用转录组基于该序列与本文所述序列的序列同源性来鉴定此类序列。参见Yinu等人(2012)Plos One[公共科学图书馆]，7(8)：e43713。本文考虑了基于其与本文所示的整个毒素受体序列或其片段的序列同一性分离的序列。此类序列包括作为公开序列的直向同源物的序列。“直向同源物”是指衍生自共同祖先基因并且由于物种形成而在不同物种中发现的基因。当其核苷酸序列和/或其编码的蛋白序列共有如本文其他地方所定义的基本同一性时，在不同物种中发现的基因被认为是直向同源物。直向同源物的功能通常在物种间是高度保守的。因此，本文考虑了编码具有毒素受体活性的多肽的并且在严格条件下与本文公开的玉米穗虫(H.zea)毒素受体序列、或与其片段杂交的多肽的分离序列。

在基于PCR的方法中，可以设计寡核苷酸引物用于PCR反应，以从由任何目的生物体提取的cDNA或基因组DNA扩增相应的DNA序列。PCR方法包括但不限于：使用成对引物、巢式引物、单特异性引物、简并引物、基因特异性引物、载体特异性引物、部分错配引物等的方法。

简并碱基，也称为摆动，是序列内给定位置处两个或更多个不同碱基的等摩尔混合物。由于遗传密码是简并的(例如，组氨酸可以由CAC或CAT编码)，所以可以在简并位点(例如，对于组氨酸，使用CAY，其中Y＝C+T)用摆动制备寡核苷酸探针。有十一种标准摆动混合物。用于指定摆动的标准代码字母如下：R＝A+G；Y＝C+T；M＝A+C；K＝G+T；S＝C+G；W＝A+T；B＝C+G+T；D＝A+G+T；H＝A+C+T；V＝A+C+G；并且N＝A+C+G+T。

简并碱基用于产生简并探针和引物。通常将简并碱基掺入设计成与编码已知氨基酸序列的未知基因杂交的寡核苷酸探针或引物中。它们也可以用于基于相似基因之间的同源性区域设计的探针或引物，以便鉴定以前未知的直向同源物。具有摆动的寡核苷酸也可用于随机诱变和组合化学。

在杂交技术中，将全部或部分已知的核苷酸序列用作与克隆的基因组DNA片段或cDNA片段(即，基因组或cDNA文库)的群体中存在的其他相应核苷酸序列进行选择性杂交的探针。这些杂交探针可以是基因组DNA片段、cDNA片段、RNA片段、或其他寡核苷酸，并且可以用可检测基团，如³²p或任何其他可检测标记进行标记。因此，例如，可以通过基于毒素受体序列标记合成的寡核苷酸来制备用于杂交的探针。用于制备杂交用探针和用于构建cDNA及基因组文库的方法通常是本领域已知的，并且在Sambrook等人(1989)Molecular Cloning：A Laboratory Manual[分子克隆：实验室手册](第2版，Cold Spring Harbor LaboratoryPress[冷泉港实验室出版社]，普莱恩维尤，纽约)中公开。

例如，本文公开的整个毒素受体序列或其一个或多个部分可以用作能够与相应的毒素受体序列和信使RNA特异性杂交的探针。为了在多种条件下实现特异性杂交，此类探针包括在毒素受体序列中是独特的并且长度为至少约10个核苷酸或长度至少约20个核苷酸的序列。此类探针可用于通过PCR对来自所选植物生物体的相应毒素受体序列进行扩增。可以使用这种技术从所需的生物体中分离另外的编码序列，或作为用于确定生物体中存在编码序列的诊断测定。杂交技术包括铺板的DNA文库的杂交筛选(斑块或群落；参见例如Sambrook等人(1989)Molecular Cloning：A Laboratory Manual[分子克隆：实验室手册](第2版，Cold Spring Harbor Laboratory Press[冷泉港实验室出版社]，Plainview[普莱恩维尤]，纽约))。

此类序列的杂交可以在严格条件下进行。“严格条件”或“严格杂交条件”是指探针与其靶序列杂交的程度将比它与其他序列杂交的程度可检测地更高(例如，比背景高至少2倍)的条件。严格条件是序列依赖性的，并且在不同情况下将有所不同。通过控制杂交和/或洗涤条件的严格性，可以鉴定与所述探针100％互补的靶序列(同源探测)。可替代地，可以调节严格条件以允许序列中的一些错配，以便检测到更低程度的相似性(异源探测)。通常，探针长度小于约1000个核苷酸，如长度小于500个核苷酸。

典型地，严格条件是以下条件，在这些条件下所述盐浓度在pH 7.0至8.3时是小于约1.5M Na⁺离子、典型为约0.01至1.0M Na⁺钠离子浓度(或其他盐)，并且对于短探针(例如，10至50个核苷酸)的温度为至少约30℃，而对于长探针(例如，超过50个核苷酸)的温度为至少约60℃。添加去稳定剂如甲酰胺也可以实现严格条件。示例性低严格条件包括在37℃用30％至35％甲酰胺、1M NaCl、1％SDS(十二烷基硫酸钠)的缓冲溶液杂交，并且在50℃至55℃在1X至2X SSC(20X SSC＝3.0M NaCl/0.3M柠檬酸三钠)中洗涤。示例性中严格条件包括在37℃在40％至45％甲酰胺、1.0M NaCl、1％SDS中杂交，并且在55℃至60℃在0.5X至1X SS℃中洗涤。示例性高严格条件包括在37℃在50％甲酰胺、1M NaCl、1％SDS中杂交，并且在60℃至65℃在0.1X SSC中洗涤。杂交的持续时间通常少于约24小时，经常为约4小时至约12小时。

特异性典型地取决于杂交后洗涤的功能，关键因素是最终洗涤溶液的离子强度以及温度。对于DNA-DNA杂交物，T_m可以从Meinkoth和Wahl(1984)Anal.Biochem.[分析生物化学]138：267-284的等式中进行估计：T_m＝81.5℃+16.6(log M)+0.41(％GC)-0.61(％form)-500/L；其中M为单价阳离子的摩尔浓度，％GC为DNA中鸟苷和胞嘧啶核苷酸的百分比，％form为杂交溶液中甲酰胺的百分比，并且L为杂合体的碱基对长度。T_m是50％的互补靶序列与完全匹配的探针杂交的温度(在所限定的离子强度和pH下)。对于每1％的错配，T_m降低约1℃；因此，可调整T_m、杂交和/或洗涤条件以与所希望的同一性的序列杂交。例如，如果查找具有＞90％同一性的序列，该T_m可以降低10℃。通常，选择严格条件为在限定离子强度和pH下低于特定序列及其互补序列的热熔点T_m约5℃。然而，极严格条件能够在比热熔点(Tm)低1℃、2℃、3℃、或4℃下杂交和/或洗涤；中严格条件能够在比热熔点(T_m)低6℃、7℃、8℃、9℃、或10℃下杂交和/或洗涤；低严格条件能够在比热熔点(T_m)低11℃、12℃、13℃、14℃、15℃、或20℃下杂交和/或洗涤。使用方程、杂交和洗涤组合物以及所需的T_m，普通技术人员将理解，本质上描述了杂交和/或洗涤溶液的严格性的变化。如果所希望的错配程度导致T_m小于45℃(水溶液)或32℃(甲酰胺溶液)，则优选增加SSC浓度以使得可使用较高温度。对核酸杂交的全面指导见于以下文献：Tijssen(1993)Laboratory Techniques inBiochemistry and Molecular Biology-Hybridization with Nucleic Acid Probes[生物化学与分子生物学技术-与核酸探针的杂交]，第I部分，第2章(Elsevier[爱思唯尔]，纽约)；以及Ausubel等人编辑.(1995)Current Protocols in Molecular Biology[分子生物学当前方案]，第2章(Greene Publishing and Wiley-Interscience[格林出版与威利交叉科学出版社]，纽约)。参见Sambrook等人(1989)Molecular Cloning：A Laboratory Manual[分子克隆：实验室手册](第2版，Cold Spring Harbor Laboratory Press[冷泉港实验室出版社]，Plainview[普莱恩维尤]，纽约)。

因此，本文涵盖了编码毒素受体蛋白并且在严格条件下与本文公开的毒素受体序列或其片段杂交的分离的序列。

该实施例的组合物和筛选方法可用于鉴定表达毒素受体的细胞、其变体和同系物。这种鉴定可以利用蛋白质水平(如配体-受体结合)或核苷酸水平的检测方法。可以通过组织切片的原位杂交原位检测多肽，但也可以通过大量多肽纯化和随后通过蛋白质印迹的分析或大量制剂的免疫学测定来分析多肽。可替代地，受体基因表达可以在核酸水平下通过任何技术领域的普通技术人员已知的技术使用互补多核苷酸来评估基因组DNA、mRNA等的水平来进行检测。作为实例，可以使用与目的核酸互补的PCR引物来鉴定表达水平。被鉴定为表达实施例的受体序列的组织和细胞被确定为对结合受体多肽的毒素是易感的。

当被鉴定为表达受体多肽的细胞来源是生物体，例如昆虫植物有害生物时，确定生物体对能够结合多肽的毒素是易感的。在一个具体实施例中，鉴定是在表达本文所示的毒素受体的鳞翅目或鞘翅目植物有害生物中进行的。

该实施例涵盖对受体多肽、修饰或编辑的受体多肽或其变体或片段具有特异性的抗体制剂。在另外的实施例中，抗体用于检测细胞中的受体表达。

在一方面，该实施例涉及用于调节植物有害生物对Bt或非Bt毒素的易感性的组合物和方法。然而，认识到这些方法和组合物可用于调节任何细胞或生物体对毒素的易感性。“调节”是指细胞或生物体对毒素的细胞毒性作用的易感性增加或降低。“易感性”是指与毒素接触的细胞的活力降低。因此，该实施例涵盖表达细胞表面受体多肽以增加或降低靶细胞或器官对毒素的易感性。毒素易感性的这种增加对于医疗和兽医学目的是有用的，在该医疗和兽医学目的中需要消除或减少一组细胞的活力。易感性的这种增加对于农业应用也是有用的，在该农业应用中需要消除或减少特定植物有害生物的群体。

目的植物有害生物包括但不限于昆虫，线虫等。线虫包括寄生线虫如根结线虫、胞囊线虫、和腐线虫，包括异皮线虫属物种、根结线虫属物种、和球异皮线虫属物种；特别是胞囊线虫的成员，包括但不限于：大豆异皮线虫(Heterodera glycines)(大豆胞囊线虫(soybean cyst nematode))；甜菜异皮线虫(Heterodera schachtii)(甜菜胞囊线虫(beetcyst nematode))；禾谷孢囊线虫(小麦禾谷孢囊线虫)；以及马铃薯金线虫(Globoderarostochiensis)和马铃薯白线虫(Globodera pailida)(马铃薯胞囊线虫)。腐线虫包括短体线虫属物种(Pratylenchus spp)。

在一个实施例中，这些方法包括产生遗传编辑或修饰的昆虫或其群落。靶受体的多核苷酸序列可以用于通过本领域技术人员已知的手段来敲除或突变昆虫中的靶受体多核苷酸，这些手段包括但不限于使用Cas9/CRISPR系统、TALEN、同源重组、和病毒转化。参见Ma等人(2014)，Scientific Reports[科学报告]，4：4489；Daimon等人(2013)，Development，Growth，and Differentiation[发育，生长和分化]，56(1)：14-25；以及Eggleston等人(2001)BMC Genetics[BMC遗传学]，2：11。

靶标受体多核苷酸的敲除或突变应该可能导致昆虫具有对杀昆虫毒素或其他杀有害生物剂降低或改变的易感性。在一些实施例中，编辑或修饰靶向基因的编码区域。在另一个实施例中，编辑或修饰靶向调节元件，例如，启动子区域。所产生的抗性昆虫或其群落可用于筛选潜在的新的活性毒素或新的或不同的作用地点的其他试剂。目前的或新颖的杀昆虫毒素也可以使用抗性昆虫系进行表征，例如用于评估目前的或新颖的杀昆虫毒素的作用方式或作用部位。

在一个实施例中，可以将如SEQ ID NO：1中所示的一种或多种天然多核苷酸或包含SEQ ID NO：1中所示序列的表达构建体以及包含所述序列的组合物通过使用双链断裂诱导剂进行基因组编辑(如CRISPR/Cas9系统)来在昆虫或昆虫细胞中进行编辑或者插入。在一个实施例中，可以将SEQ ID NO：l中所示的基因组DNA序列通过使用双链断裂诱导剂进行基因组编辑(如CRISPR/Cas9系统)来进行编辑或插入。如本文所用，术语“编辑的”或“遗传编辑的”意指使用双链或单链断裂诱导剂(如Cas9/CRISPR系统)来诱导天然序列的变化。

CRISPR基因座(规律间隔成簇短回文重复序列)(又称SPIDR--间隔区散在同向重复序列)构成最近描述的DNA基因座的家族。CRISPR基因座由部分回文的短而高度保守的DNA重复序列(通常为24至40bp，重复从1至140次，也称为CRISPR重复序列)组成。重复序列(通常具有物种特异性)由恒定长度的可变序列(通常为20至58，依赖于CRISPR基因座(参见WO 2007/025097))间隔。

Cas内切核酸酶涉及由Cas基因编码的Cas蛋白质，其中所述Cas蛋白质能够将双链断裂引入DNA靶序列中。Cas内切核酸酶由指导多核苷酸指导以识别并任选地在特定靶位点向细胞基因组中引入双链断裂(参见U.S.2015/0082478)。指导多核苷酸/Cas内切核酸酶系统包括能够将双链断裂引入DNA靶序列的Cas内切核酸酶和指导多核苷酸的复合物。如果正确的前间区序列邻近基序(PAM)在靶序列的3’末端适当地定向时，Cas内切核酸酶在基因组靶位点附近极为贴近处解开DNA双链体，并且在通过指导RNA识别靶序列时切割两条DNA链。

在一个实施例中，这些方法包括产生昆虫细胞、昆虫或其群落，其中靶基因被编辑，使得其不再起作用。靶基因的多核苷酸序列可以用于通过本领域技术人员已知的手段来敲除昆虫中的靶基因多核苷酸，这些手段包括但不限于使用Cas9/CRISPR系统、TALEN、同源重组、和病毒转化。参见Ma等人(2014)，Scientific Reports[科学报告]，4：4489；Daimon等人(2013)，Development，Growth，and Differentiation[发育，生长和分化]，56(1)：14-25；以及Eggleston等人(2001)BMC Genetics[BMC遗传学]，2：11。

在一个实施例中，这些方法涉及导致通过靶受体多核苷酸表达(例如，通过RNAi靶向负调节元件)实现的抗性挽救或逆向突变的方法。

在一个实施例中，这些方法涉及产生对至少一种杀昆虫毒素有抗性的昆虫群落。群落可以通过遗传修饰或编辑方法产生。可替代地，受体多核苷酸可用于筛选突变体、缺乏靶受体多核苷酸的昆虫或任何其他遗传变体。随后对杀昆虫毒素的筛选和选择应该导致可以如本文所述使用的抗性群落。方法包括但不限于：喂养昆虫来自玉蜀黍植物(这些玉蜀黍植物表达施用于人工饲料的杀昆虫剂或纯化的杀昆虫剂)的叶物质，并且选择暴露后幸存的个体。这些方法进一步可以涉及将存活的昆虫转移到缺乏杀昆虫剂的标准饲料中，以使幸存者完成发育。这些方法进一步可以涉及通过喂养昆虫来自玉蜀黍植物(这些玉蜀黍植物表达杀昆虫剂或纯化的杀昆虫剂)的叶物质允许存活的昆虫交配来维持群落(其中选择周期性施用于后代)并选择存活的昆虫，并且因此通过消除不携带纯合抗性等位基因的个体来确定抗性。

本文公开的方法和组合物涉及遗传编辑的昆虫细胞、昆虫或昆虫群落，其中所述遗传编辑的昆虫细胞、昆虫或昆虫群落对杀昆虫毒素具有抗性。在一些实施例中，在任何遗传编辑之前，昆虫细胞、昆虫或昆虫细胞群落对杀昆虫毒素敏感。在一些实施例中，天然杀昆虫毒素受体是遗传编辑的。在某些实施例中，天然杀昆虫毒素受体包含SEQ ID NO：2中所示的序列。

一个实施例涵盖用于筛选对杀昆虫剂具有改变的易感性水平的昆虫群体的方法，包括抗性监测测定。用于筛选改变的易感性水平的测定包括但不限于：测定靶受体基因DNA序列、RNA转录物、多肽、或靶受体多肽的活性。测定方法包括但不限于：DNA测序、DNA印迹法、RNA印迹法、RNA测序、PCR、RT-PCR、qPCR、qRT-PCR、蛋白质测序、蛋白质印迹、质谱鉴定、抗体制备和检测、以及酶检测。DNA、RNA转录物、或多肽中的序列变化可以指示抗性昆虫。此外，目标受体多肽的RNA、多肽、或酶活性的量或丰度的变化可以指示抗性昆虫。在一个实施例中，该方法包括在选择下筛选昆虫以增加受体介导的抗性的选择效率。在另一个实施例中，该方法包括筛选SEQ ID NO：2的公开的多肽受体中的突变或改变的序列、SEQ ID NO：2的表达的变化、或SEQ ID NO：1的表达的变化、或其互补序列，其中该变化指示受体介导的毒素抗性。在另一个实施例中，所述方法涉及针对不同于天然非抗性昆虫序列的受体基因或基因产物、转录物、或多肽序列筛选昆虫。在一个实施例中，将具有改变或突变序列的昆虫进一步暴露于杀昆虫毒素，其中所述杀昆虫毒素具有与Bt毒素相同的作用位点。筛选受体的使用允许进行有效的受体介导的抗性选择以产生抗性昆虫群落。

在一个实施例中，所述方法涉及用于在田间监测对杀昆虫毒素的昆虫抗性或改变的易感性水平的方法，所述方法包括测定改变的易感性水平或昆虫抗性，其可以包括但不限于：测定靶受体基因DNA序列、RNA转录物、多肽、或靶受体多肽的活性。测定方法包括但不限于：DNA测序、DNA印迹法、RNA印迹法、RNA测序、PCR、RT-PCR、qPCR、qRT-PCR、蛋白质测序、蛋白质印迹、质谱鉴定、抗体制备和检测、或酶检测。DNA、RNA转录物、或多肽中的序列变化可以指示抗性昆虫。此外，目标受体多肽的RNA、多肽、或酶活性的量或丰度的变化可以指示抗性昆虫。在另一个实施例中，该方法包括筛选SEQ ID NO：2的公开的多肽受体中的突变或改变的序列、SEQ ID NO：2的表达的变化、或SEQ ID NO：1的表达的变化、或其互补序列，其中该变化指示受体介导的毒素抗性。在另外的实施例中，该方法涉及将杀昆虫剂应用于具有不同于天然序列的受体基因或基因产物序列的昆虫或昆虫群体环境周围的区域，其中与杀昆虫毒素相比，所述杀昆虫剂具有不同的作用方式。在另外的实施例中，该方法包括实施昆虫管理抗性(IRM)计划或有害生物综合管理计划(IPM)。在一个实施例中，IPM计划可以包括但不限于：添加避难所或另外的避难所、轮作作物、种植另外的自然避难所、以及在不同的作用位点应用杀昆虫剂。

在一个实施例中，这些方法包括监测抗性的测定试剂盒。简单的试剂盒可以在田间或实验室中使用，以筛选存在的抗性昆虫。在优选的实施例中，针对SEQ ID NO：2产生的抗体可用于确定昆虫群体中SEQ ID NO：2的水平、或其存在或不存在、或其浓度变化。在另一个实施例中，进行SEQ ID NO：1的评估，以评估昆虫或昆虫群体靶受体序列中的序列变化、或相对于对照的表达变化、或序列变异。用来完成试剂盒中的抗性监测的分子技术，例如但不限于：PCR、RT-PCR、qRT-PCR、DNA印迹法、RNA印迹法、及其他。

以下实例是通过说明的方式但不是通过限制的方式来提供的。

实例

实例1：DvABCB1序列在易感性WCR转录组中的鉴定。

基于ABCB1为叶甲(白杨叶甲(Chrysomela temula))中Cry3Aa的功能性受体的报道，鉴定WCR中的推定的ABCB1直系同源物以表征和理解其是否在介导Cry3毒性中发挥相似作用(Pauchet等人(2016)，Toxins[毒素]8(12))。针对WCR转录组进行的CtABCB1序列的BLAST(tblastx)检索鉴定了在氨基酸水平上具有69％总体同一性的序列(Davis-Vogel C等人(2018)，PloS One[公共科学图书馆]13(9)：e0203160)。命中率最高的Dvv-isotig11620编码1256个氨基酸的蛋白质，该蛋白质被命名为DvABCB1，预测大小为138kDa。

实例2：DvABCB1在Sf9和HEK293细胞中的异源表达为Cry3A毒素赋予细胞毒性。

已使用基于细胞的测定来评估昆虫毒素受体的功能性作用(Soberon M等人(2018)Insect Biochem Mol Biol[昆虫生物化学与分子生物学]93：66-78)。我们用表达DvABCB1的重组杆状病毒以高滴度感染培养中的Sf9细胞，以研究和表征随后添加Cry毒素对Sf9细胞的毒性/活力的影响。所测试的毒素包括Cry3Aa、Cry34Ab1/35Ab1和Cry6Aa1，它们都被制备并激活。(Li H等人(2013)PloS One[公共科学图书馆]8(1)：353079；Fortea等人(2017)J Biol Chem[生物化学杂志])。在与激活的毒素一起温育过夜后，通过相差光学显微镜评估细胞的形态变化。对于暴露于Cry3Aa处理的细胞，观察到几乎完全的细胞死亡，但对于暴露于Cry34Ab1/Cry35Ab1处理或Cry6Aa1处理的细胞，没有观察到任何影响。细胞形态与仅暴露于培养基的对照细胞相似。这些结果表明，DvABCB1在Sf9细胞中的异源表达选择性地对Cry3Aa赋予细胞毒性，但不对Cry34Ab1/35Ab1或Cry6Aa1赋予细胞毒性。

为更定量地表征DvABCB1的功能性，将DvABCB1在HEK293细胞中表达以建立稳定的细胞系。DvABCB1 cDNA的表达载体包括荧光标签以帮助克隆和表达的表征。当在共焦显微镜下检查时，在HEK293细胞的细胞表面上观察到来自DvABCB1-RFP的荧光。用增加剂量的Cry3Aa样毒素IP3-H9处理表达DvABCB1-RFP的细胞导致剂量依赖性细胞死亡，同时，未转染细胞对IP3-H9暴露无应答。IP3-H9是与Cry3A相比具有改善的溶解度的Cry3A的修饰形式，并且在人工饲料生物测定中对Cry3A具有完全交叉抗性(Bermudez等人(2015)，美国专利9109231 B2；Wei JZ等人(2018)Plant Biotechnol J[植物生物技术杂志]16(2)：649-659；Zhao JZ等人(2016)J Ecin Entomol[经济昆虫学杂志])。定量细胞应答显示了在100nM毒素处理下约70％的细胞死亡率，其中半数最大应答的浓度为大约0.68nM(EC50值)。其余30％的代谢活性细胞可能不表达足够的DvABCB1受体以应答于毒素暴露。这些结果与先前在Sf9细胞测定中报道的观察结果相似(Pauchet Y等人(2016)Toxins(Basel)[毒素(巴塞尔)]8(12)；Bretschneider A等人(2016)Insect Biochem Mol Bio[昆虫生物化学与分子生物学]76：109-117)，并且证明当在HEK293细胞中表达时，来自WCR的DvABCB1也可以作为Cry3A受体。

实例3：通过RNAi敲低DvABCB1使得WCR幼虫对Cry3A样毒素IP3-H9不敏感。

使用RNAi抑制DvABCB1的表达并证明随后的经处理的WCR对Cry3A暴露不敏感进一步验证了DvABCB1在对WCR幼虫的Cry3A毒性中的功能性作用。设计155bp的双链RNA(dsRNA)以靶向DvABCB15’端附近的区域。用24小时大的WCR幼虫使用两阶段人工饲料生物测定来进行测定。在第1阶段(长4天)，将幼虫暴露于高剂量的dsRNA(100μg/mL饮食)以有效地沉默DvABCB1。在第2阶段期间，将这些幼虫转移到新鲜饲料中，并暴露于WCR活性毒素10天。在第1阶段，对照包括对应于GUS(β-葡糖醛酸酶)的dsRNA和单独的水。此外，将幼虫暴露于dsRNA2天，并且然后转移到新鲜的含dsRNA的饲料中再暴露2天，以减少饲料中dsRNA降解的影响。使用qRT-PCR来评价在第1阶段期间通过暴露于DvABCB1 dsRNA 4天所实现的沉默效率。qRT-PCR结果显示实现了与验证第1阶段的暴露条件的水和GUS dsRNA处理所观察到的转录物水平相比，超过90％的DvABCB1转录物抑制，这表明其被非常有效的抑制。第2阶段处理包括分别暴露于IP3-H9、IPD072Aa、缓冲液和水对照。IPD072Aa是一种新的WCR活性毒素，其对mCry3Aa或Cry34Abl/Cry35Ab1抗性WCR幼虫没有交叉抗性(Schellenberger U等人(2016)Science[科学]354(6312)：634-637)。进行初步生物测定以表征未处理幼虫在第2阶段暴露期间对IP3-H9和IPD072Aa的敏感性，以确定用于毒性测试的适当剂量。第2阶段暴露于IP3-H9(200μg/mL)对在第1阶段预暴露于GUS dsRNA或水对照的幼虫造成约50％死亡率和90％生长抑制作用。类似地，第2阶段暴露于IPD072Aa对在第1阶段预暴露于GUS dsRNA或水对照的幼虫造成高死亡率(76％-86％)和生长抑制(98％-99％)。第2阶段阴性对照的死亡率和生长抑制低于5％，表明先前暴露于DvABCB1或GUS dsRNA不影响幼虫生长和发育。

实例4.Cry3A抗性玉米根萤叶甲中的DvABCB1转录物的改变的鉴定。

遵循RNeasy方案与任选的柱上DNA酶消化(optional on-column DNasedigestion)(凯杰公司(Qiagen)，希尔登，德国)，将全部RNA从冷冻肠道中分离，这些肠道是从易感性和mCry3A抗性3龄昆虫中提取的(10根肠道为一组，每组3个样品)。为获得用于测序的扩增子，使用用SuperScriptIII逆转录酶(生命科技公司(Life Technologies)，卡尔斯巴德，加利福尼亚州)生成的cDNA进行RT-PCR。使用Invitrogen Platinum SuperFi PCR预混合物(生命科技公司，卡尔斯巴德，加利福尼亚州)使用来自每种菌株的cDNA(DvABCB1FwdStrt_5’-ATGACAGAAGAAAAAAAACATAGTATAAAGG-3’和DvABCB1RevStop_5’-TTACGTTTTTTGAGTATATAATTTGTAGTACAG-3’)来设置用于DvABCB1的PCR反应。在文库制备前，将所得的PCR扩增子使用Ampure PB珠来纯化和浓缩。使用太平洋生物科学公司(PacificBioscience)的扩增子条码试剂盒(Amplicon Barcoding kit)，将独特条形码化接头与每个扩增子连接。在合并之后，使用Sequel结合试剂盒3.0将文库与测序引物和聚合酶结合。使用Sequel测序试剂盒3.0将其加载到Sequel上的单个样品室上，运行配置为20小时。使用ccs应用(4.0.0版，太平洋生物科技公司(Pacific Biosciences，Inc.))生成CCS读数，minPasses＝3，并且minPredictedAccuracy＝0.999。提取以正确的独特条码和引物序列开头的序列。分析每个样品的出现率大于5％的序列。这些序列为来自Cry3A抗性WCR样品的DvABCB1_3AR1、DvABCB1_3AR_2、DvABCB1_3AR3和来自易感性WCR样品的DvABCB1 Sus1。将这四个核苷酸序列使用Geneious Prime进行比对(图1)。在DvABCB1_3AR1和DvABCB1_3AR2中分别鉴定出726bp和2430bp的大缺失，没有移码或终止密码子。由于24个SNP，DvABCB1_Sus1和DvABCB1_3AR3共享99.36％的同一性。

使用WCR基因组模型序列(NCBI Dvir_v2.0；登录号NW_021043569)和经鉴定的DvABCB1核苷酸序列(NCBI中DvABCB1的TBLASTN)比对3AR1和3AR2序列，揭示了缺失序列的开始发生在LOC114344372的注释外显子7之后，在外显子-内含子接合处。DvABCB1_3AR1序列在LOC114344372的注释外显子11(模型中的最后一个注释外显子)的开始处恢复。NW_021043569仅含有DvABCB1的部分序列(该基因序列的后半部分不存在)并且不允许鉴定DvABCB1_3AR2序列恢复处的外显子。无论如何，外显子-内含子接合处的缺失的位置提供了抗性序列为可变剪接产物的强烈迹象。对于Cry3抗性的机制来说更重要的是，预计该缺失的程度会消除对折叠和膜拓扑结构有显著影响的蛋白质关键区域，从而对Cry3蛋白质的结合有不利影响。

实例5.DvABCB1的定量实时PCR(qRT-PCR)。

在拉出的3龄WCR幼虫的肠道中定量DvABCB1基因的表达，每个样品10根肠道。引物和探针区域的设计在表1中列出。使用一步实时定量RT-PCR分析基因表达。使用单重设置用Bioline Sensifast Probe Lo Rox试剂盒(陶顿，马萨诸塞州)进行测定，每个样品重复2次，并使用2-^ΔΔCt方法基于靶基因和参考基因DvRPS10(GenBank登录号KU756281)的相对表达来进行分析。表2示出了比较易感性和Cry3抗性WCR中的DvABCB1表达水平的表达分析结果。

表1.用于qRT-PCR测定的引物和探针

表2.通过5’端和del(Res群落中的缺失区域)的qRTPCR测定的WCR易感性(Sus)和抗性(Res)菌落中的DvABCB1转录物。

实例6：WCR中受体的鉴定。

使用内部注释管道注释从Cry34/35易感性个体组装的基因组草图。除了从头预测之外，转录组数据还用作基因模型预测的证据。分析具有一个或多个ABC相关的Pfam结构域的基因模型是否存在跨膜结构域。将WCRW的AF252中肠样品(8个重复)与基因组草图进行比对，并获得归一化读数计数。将满足以下3项要求的基因列为ABC受体-(1)含有ABC相关的Pfam结构域，(2)在基因表达研究中使用的五个或更多个样品中非零表达，和(3)10个或更多个跨膜结构域。将FAW受体序列与列出的ABC受体进行比较以鉴定WCRW中的直系同源基因。将具有高查询覆盖率和百分比同一性的Blast命中优先考虑为潜在的受体。鉴定的受体在表3中示出。

表3.WCR受体

本说明书中提到的所有出版物和专利申请都指示了本公开所属领域的技术人员的水平。将所有出版物和专利申请通过引用并入本文，其程度就像明确且单独指出通过引用将每个单独出版物或专利申请并入本文一样。

虽然为了清楚理解的目的，已经通过说明和实例的方式对前述实施例进行了详细描述，但是在所附权利要求的范围内可以实施某些改变和修改。

序列表

<110> 先锋良种国际有限公司（Pioneer Hi-Bred International, Inc.）

Samantha, Griffin

<120> 新颖的杀昆虫毒素受体和使用方法

<130> 8424

<160> 63

<170> PatentIn 3.5版

<210> 1

<211> 3771

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 1

atgacagaag aaaaaaaaca tagtataaag gataaagaga aaattggtat tgatgcccaa 60

tttgttaata gtgaagaacc aaaggaaaaa attaagaatg tatcttttcc tcagatgttt 120

aggtatgcaa gtacttatga taaattttta atggtagtag gacttatatc ggcaactgga 180

acaggagtct tacaacccct aaatacgatt ctctttggta gcctcacagg agatatcatt 240

gcatatgcta cgtcgataca gataaattta cctgccgatc agaagaaaat agcagaagac 300

aatttttttg atggtatccg atattttgcc ctaatgaatt ctctgatcgg aataggaatg 360

tttgtgttca gttacctagc aacagtcact tttaactatt ctgctatgag acagatattc 420

aaaataaggt cagcatattt gaaatccatt ttaaaccaag atgtaggttg gtacgatatt 480

aatcagactg gagattttgc cagtaggatg tcagatgatt tgttcaaatt tgaagatggc 540

ataggtgaaa aagtaccggt attctggagt tttcaagttg tttttttaac ttcacttatc 600

atagctcttg tgaaaggatg ggaacttgct ttaatatgtt taacctctct acccgctaca 660

ttaataacaa tcggaattgt aggacttctc actacaaaat tagcaaaaaa tgaactggaa 720

gcatatggcg ctgctggatc aatagccgaa gaagcattgt ctttaattag aaccataacg 780

gcttttggag gtcagaagaa agaagtagac agatataata aaaatttagt tgaagccaaa 840

aacaataata taagaagatc gatgttttca gcattaggtt tcggtttatt gtggttcatg 900

atctatgcca gttatgctct ggcattctgg tatggtgtaa aattagtttt aagggacagg 960

acagctacga atcaaatata tactcctagt aatatggtga ctgttttctt tagtgtcatg 1020

acaggatcta tgaactttgg tattgcctct ccatatatag aggcatttgg aatctcaaaa 1080

gcggcggcgt caaaaatttt tagtgtaatc gatcacaaac ctacaattaa tttatcaaaa 1140

ggaaatggaa aaacgctaaa cgtactcata ggaaatatac aatttaaaga tgtggctttc 1200

cgttatccat ctaggaaaga tgtaccgata ttaaaaggat taagtttgaa cattaaatct 1260

ggagacactg tggcacttgt aggtagttcg ggctgtggaa aatctacggt tattcagtta 1320

cttcaacgat tatacgatgc ggactctgga gaggtcacca tagacggaaa aaatattaaa 1380

gaatacgatt taacttggtt aagaagccaa ataggagttg taggtcaaga accaattctc 1440

tttggaacat ccattttaga aaatattagg tacggtaaag atggagtaac agaggaagat 1500

gtcattcaag cagctaaaaa agctaatgcg cataacttta ttaaagctct tcctaacgga 1560

tataatacat tggttggaga gaaaggggct caactctcag gtggacaaaa acaaaggata 1620

gctatagcta gagctttggt aagaaaccct actctactgc ttttggacga agctacttca 1680

gcactggata ataccagtga agctaaagta caagcagctc tagatgcggc aagtgttgaa 1740

tgtacaacta taattgtggc ccacaggcta tcaacaattc gaggtgcaaa caaaattatc 1800

gtactttctc aaggagtcgt ggtagaagaa ggaacccacg aagaactaat ggaacttaaa 1860

caagagtact atagattagt aacagcccaa gtcaaatcat cggaacagtt tgaagtcgct 1920

gaaaagaaga aagtggtaag agctattagc ttggcagaaa gtagcacagg ctctgaccat 1980

aacatagagg ctacaaaaga agataatgaa gacgatttta atgaaaataa agacgtttcc 2040

gtttttgaaa tattaaagat gaatgctcca gaatggcctt acattttatt tgctggtctt 2100

ggatcaattg ttgttggttg tggaatgccc gtttttgcag tcttgtttgg tagcattttg 2160

gggacattag caaatggtga tcctgacttt gtaagatctg aaaccaacaa atattgctta 2220

tattttgtgc tgggaggact cataactatg gtatcagtat ttactcagat gtacctgtta 2280

ggaattgctg gtgaaaaaat gacagaaaga gttaggagtc gattattcaa ggcaatgata 2340

taccaggaaa taggcttctt tgacaagaaa acaaatgggg ttggagcgct ttgtgccaaa 2400

ttgtcttctg atgcttctaa tattcaaggg gcaacaggaa ttcgggttgg aaccattctt 2460

caatcaatag ccacattttg cttagcaatt ggtctttcca tgtattatga atggaaattg 2520

ggtttagtta cagcagcttt cacacctgtt attttgatcg ctatgttctt tgaaagaaga 2580

aacaccagag gtggaaacga ttcaagagat tccgcactac aaaaatcaac aaggactgca 2640

gtagaagccg ttgggaatat acgaacggtt gcctccttgg gcttagaaga aaagttccag 2700

caactatatg aatctgagct catgccacat tataaatctt cgttaaaaac agttcactgg 2760

agagcaatcg tgtttggtct ttccagaagt ttgttgttct ttgcttatgc tactgcaatg 2820

tattacggcg gatttttgat aagggatgga ttgccatacg atagagtatt taaggtatct 2880

caagcacaaa ttatgggtac agtttcaatt gccaactcac tggcattttc tccaaacttt 2940

gctaagggtg ttgcagcagc aaaaaaagtt aaaagcttcc tttcaagaat tccacttatt 3000

cgtgatttac cttcttcacg acaaatggtt aaggcaagtg gcaacttcag tttttcagaa 3060

atcgaattta cgtatccaac acgacccaac gtactaattt taaaaggact taatttggac 3120

attctaaatg gaaaaactgt agctctagtg ggagaaagtg gttgcggaaa atctacgata 3180

attcaactca tcgagagatt ttacgatcct agatctggtg aagtaaaaat ggatggtgta 3240

gatttaaaag atatatcact agattcgttg aggtcgcata tgggtatagt atcacaagaa 3300

ccaaatctgt tcaataagtc tattgctgaa aatattgctt atggagataa ttcaagggag 3360

gtttctatgg atgagataat taaagctgct aaaaatgcca atattcataa ttttataact 3420

ggattaccga agggatatga aacaaaacta ggagaaaagg ctgttcagtt gtctggtgga 3480

caaaaacaaa gaattgctat tgccagagcg ttagtaagaa atccaaaagt tttactcctt 3540

gatgaagcca catctgcact agacactgaa agtgagaagg ttgttcaaga agctctagat 3600

caagctaaaa aaggaaggac atgtgttacg attgcccaca gattaacaac catacaagat 3660

gcagacttga tatgcgtagt agcaaatggt gtaattgctg aatctggaag ccatcaagaa 3720

cttctgcaaa aggagggact gtactacaaa ttatatactc aaaaaacgta a 3771

<210> 2

<211> 1256

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 2

Met Thr Glu Glu Lys Lys His Ser Ile Lys Asp Lys Glu Lys Ile Gly

1 5 10 15

Ile Asp Ala Gln Phe Val Asn Ser Glu Glu Pro Lys Glu Lys Ile Lys

20 25 30

Asn Val Ser Phe Pro Gln Met Phe Arg Tyr Ala Ser Thr Tyr Asp Lys

35 40 45

Phe Leu Met Val Val Gly Leu Ile Ser Ala Thr Gly Thr Gly Val Leu

50 55 60

Gln Pro Leu Asn Thr Ile Leu Phe Gly Ser Leu Thr Gly Asp Ile Ile

65 70 75 80

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

85 90 95

Ile Ala Glu Asp Asn Phe Phe Asp Gly Ile Arg Tyr Phe Ala Leu Met

100 105 110

Asn Ser Leu Ile Gly Ile Gly Met Phe Val Phe Ser Tyr Leu Ala Thr

115 120 125

Val Thr Phe Asn Tyr Ser Ala Met Arg Gln Ile Phe Lys Ile Arg Ser

130 135 140

Ala Tyr Leu Lys Ser Ile Leu Asn Gln Asp Val Gly Trp Tyr Asp Ile

145 150 155 160

Asn Gln Thr Gly Asp Phe Ala Ser Arg Met Ser Asp Asp Leu Phe Lys

165 170 175

Phe Glu Asp Gly Ile Gly Glu Lys Val Pro Val Phe Trp Ser Phe Gln

180 185 190

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

195 200 205

Leu Ala Leu Ile Cys Leu Thr Ser Leu Pro Ala Thr Leu Ile Thr Ile

210 215 220

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

225 230 235 240

Ala Tyr Gly Ala Ala Gly Ser Ile Ala Glu Glu Ala Leu Ser Leu Ile

245 250 255

Arg Thr Ile Thr Ala Phe Gly Gly Gln Lys Lys Glu Val Asp Arg Tyr

260 265 270

Asn Lys Asn Leu Val Glu Ala Lys Asn Asn Asn Ile Arg Arg Ser Met

275 280 285

Phe Ser Ala Leu Gly Phe Gly Leu Leu Trp Phe Met Ile Tyr Ala Ser

290 295 300

Tyr Ala Leu Ala Phe Trp Tyr Gly Val Lys Leu Val Leu Arg Asp Arg

305 310 315 320

Thr Ala Thr Asn Gln Ile Tyr Thr Pro Ser Asn Met Val Thr Val Phe

325 330 335

Phe Ser Val Met Thr Gly Ser Met Asn Phe Gly Ile Ala Ser Pro Tyr

340 345 350

Ile Glu Ala Phe Gly Ile Ser Lys Ala Ala Ala Ser Lys Ile Phe Ser

355 360 365

Val Ile Asp His Lys Pro Thr Ile Asn Leu Ser Lys Gly Asn Gly Lys

370 375 380

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

385 390 395 400

Arg Tyr Pro Ser Arg Lys Asp Val Pro Ile Leu Lys Gly Leu Ser Leu

405 410 415

Asn Ile Lys Ser Gly Asp Thr Val Ala Leu Val Gly Ser Ser Gly Cys

420 425 430

Gly Lys Ser Thr Val Ile Gln Leu Leu Gln Arg Leu Tyr Asp Ala Asp

435 440 445

Ser Gly Glu Val Thr Ile Asp Gly Lys Asn Ile Lys Glu Tyr Asp Leu

450 455 460

Thr Trp Leu Arg Ser Gln Ile Gly Val Val Gly Gln Glu Pro Ile Leu

465 470 475 480

Phe Gly Thr Ser Ile Leu Glu Asn Ile Arg Tyr Gly Lys Asp Gly Val

485 490 495

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

500 505 510

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

515 520 525

Gly Ala Gln Leu Ser Gly Gly Gln Lys Gln Arg Ile Ala Ile Ala Arg

530 535 540

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

545 550 555 560

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

565 570 575

Ala Ser Val Glu Cys Thr Thr Ile Ile Val Ala His Arg Leu Ser Thr

580 585 590

Ile Arg Gly Ala Asn Lys Ile Ile Val Leu Ser Gln Gly Val Val Val

595 600 605

Glu Glu Gly Thr His Glu Glu Leu Met Glu Leu Lys Gln Glu Tyr Tyr

610 615 620

Arg Leu Val Thr Ala Gln Val Lys Ser Ser Glu Gln Phe Glu Val Ala

625 630 635 640

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

645 650 655

Gly Ser Asp His Asn Ile Glu Ala Thr Lys Glu Asp Asn Glu Asp Asp

660 665 670

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

675 680 685

Ala Pro Glu Trp Pro Tyr Ile Leu Phe Ala Gly Leu Gly Ser Ile Val

690 695 700

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

705 710 715 720

Gly Thr Leu Ala Asn Gly Asp Pro Asp Phe Val Arg Ser Glu Thr Asn

725 730 735

Lys Tyr Cys Leu Tyr Phe Val Leu Gly Gly Leu Ile Thr Met Val Ser

740 745 750

Val Phe Thr Gln Met Tyr Leu Leu Gly Ile Ala Gly Glu Lys Met Thr

755 760 765

Glu Arg Val Arg Ser Arg Leu Phe Lys Ala Met Ile Tyr Gln Glu Ile

770 775 780

Gly Phe Phe Asp Lys Lys Thr Asn Gly Val Gly Ala Leu Cys Ala Lys

785 790 795 800

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

805 810 815

Gly Thr Ile Leu Gln Ser Ile Ala Thr Phe Cys Leu Ala Ile Gly Leu

820 825 830

Ser Met Tyr Tyr Glu Trp Lys Leu Gly Leu Val Thr Ala Ala Phe Thr

835 840 845

Pro Val Ile Leu Ile Ala Met Phe Phe Glu Arg Arg Asn Thr Arg Gly

850 855 860

Gly Asn Asp Ser Arg Asp Ser Ala Leu Gln Lys Ser Thr Arg Thr Ala

865 870 875 880

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

885 890 895

Glu Lys Phe Gln Gln Leu Tyr Glu Ser Glu Leu Met Pro His Tyr Lys

900 905 910

Ser Ser Leu Lys Thr Val His Trp Arg Ala Ile Val Phe Gly Leu Ser

915 920 925

Arg Ser Leu Leu Phe Phe Ala Tyr Ala Thr Ala Met Tyr Tyr Gly Gly

930 935 940

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

945 950 955 960

Gln Ala Gln Ile Met Gly Thr Val Ser Ile Ala Asn Ser Leu Ala Phe

965 970 975

Ser Pro Asn Phe Ala Lys Gly Val Ala Ala Ala Lys Lys Val Lys Ser

980 985 990

Phe Leu Ser Arg Ile Pro Leu Ile Arg Asp Leu Pro Ser Ser Arg Gln

995 1000 1005

Met Val Lys Ala Ser Gly Asn Phe Ser Phe Ser Glu Ile Glu Phe

1010 1015 1020

Thr Tyr Pro Thr Arg Pro Asn Val Leu Ile Leu Lys Gly Leu Asn

1025 1030 1035

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

1040 1045 1050

Gly Cys Gly Lys Ser Thr Ile Ile Gln Leu Ile Glu Arg Phe Tyr

1055 1060 1065

Asp Pro Arg Ser Gly Glu Val Lys Met Asp Gly Val Asp Leu Lys

1070 1075 1080

Asp Ile Ser Leu Asp Ser Leu Arg Ser His Met Gly Ile Val Ser

1085 1090 1095

Gln Glu Pro Asn Leu Phe Asn Lys Ser Ile Ala Glu Asn Ile Ala

1100 1105 1110

Tyr Gly Asp Asn Ser Arg Glu Val Ser Met Asp Glu Ile Ile Lys

1115 1120 1125

Ala Ala Lys Asn Ala Asn Ile His Asn Phe Ile Thr Gly Leu Pro

1130 1135 1140

Lys Gly Tyr Glu Thr Lys Leu Gly Glu Lys Ala Val Gln Leu Ser

1145 1150 1155

Gly Gly Gln Lys Gln Arg Ile Ala Ile Ala Arg Ala Leu Val Arg

1160 1165 1170

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

1175 1180 1185

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

1190 1195 1200

Lys Gly Arg Thr Cys Val Thr Ile Ala His Arg Leu Thr Thr Ile

1205 1210 1215

Gln Asp Ala Asp Leu Ile Cys Val Val Ala Asn Gly Val Ile Ala

1220 1225 1230

Glu Ser Gly Ser His Gln Glu Leu Leu Gln Lys Glu Gly Leu Tyr

1235 1240 1245

Tyr Lys Leu Tyr Thr Gln Lys Thr

1250 1255

<210> 3

<211> 3045

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 3

atgacagaag aaaaaaaaca tagtataaag gataaagaga aaattggtat tgatgcccaa 60

tttgttaata gtgaagaacc aaaggaaaaa attaagaatg tatcttttcc tcagatgttt 120

aggtatgcaa gtacttatga taaattttta atggtagtag gacttatatc ggcaactgga 180

acaggagtct tacaacccct aaatacgatt ctctttggta gcctcacagg agatatcatt 240

gcatatgcta cgtcgataca gataaattta cctgccgatc agaagaaaat agcagaagac 300

aatttttttg atggtatccg atattttgcc ctaatgaatt ctctgatcgg aataggaatg 360

tttgtgttca gttacctagc aacagtcact tttaactatt ctgctatgag acagatattc 420

aaaataaggt cagcatattt gaaatccatt ttaaaccaag atgtaggttg gtacgatatt 480

aatcagactg gagattttgc cagtaggatg tcagatgatt tgttcaaatt tgaagatggc 540

ataggtgaaa aagtaccggt attctggagt tttcaagttg tttttttaac ttcacttatc 600

atagctcttg tgaaaggatg ggaacttgct ttaatatgtt taacctctct acccgctaca 660

ttaataacaa tcggaattgt aggacttctc actacaaaat tagcaaaaaa tgaactggaa 720

gcatatggcg ctgctggatc gatagccgaa gaagcattgt ctttaattag aaccataacg 780

gcttttggag gtcagaagaa agaagtagac agatataata aaaatttagt tgaagccaaa 840

aacaataata taagaagatc gatgttttca gcattaggtt tcggtttatt gtggttcatg 900

atctatgcca gttatgctct ggcattctgg tatggtgtaa aattagtttt aagggacagg 960

acagctacga atcaaatata tactcctagt aatatggtga ctgcaagtgt tgaatgtaca 1020

actataattg tggcacacag gctatcaaca attcgaggtg caaacaaaat tatcgtactt 1080

tctcaaggag tcgtggtaga agaaggaacc cacgaagaac taatggaact taaacaagag 1140

tattatagat tagtaacagc ccaagtcaaa tcatcggaac agtttgaagt cgctgaaaag 1200

aagaaagtgg taagagctat tagcttggca gaaagtagca caggctctga ccataacata 1260

gaggctacaa aagaagataa tgaagacgat tttaatgaaa ataaagacgt ttccgttttt 1320

gaaatattga agatgaatgc tccagaatgg ccttacattt tatttgctgg tcttggatca 1380

attgttgttg gttgtggaat gcccgttttt gcagtcttgt ttggtagcat tttggggaca 1440

ttagcaaatg gtgatcctga ctttgtaaga tctgaaacca acaaatattg cttatatttt 1500

gtgctgggag gactcataac tatggtatca gtatttactc agatgtacct gttaggaatt 1560

gctggtgaaa aaatgacaga aagagttagg agtcgattat tcaaggcaat gatataccag 1620

gaaataggct tctttgacaa gaaaacaaat ggggttggag cgctttgtgc caaattgtct 1680

tctgatgctt ctaatattca aggggcaacg ggaattcggg ttggaaccat tcttcaatca 1740

atagccacat tttgcttagc aattggtctt tccatgtatt atgaatggaa attgggctta 1800

gttacagcag ctttcacacc tgttattttg atcgctatgt tctttgaaag aagaaacacc 1860

agaggtggaa acgattcaag agattccgca ctacaaaaat caacaaggac tgcagtagaa 1920

gccgttggga atatacgaac agttgcctcc ttaggcttag aagaaaagtt ccagcagcta 1980

tatgaatctg agctcatgcc acattataaa tcttcgttaa aaacagttca ctggagagca 2040

atcgtgtttg gtctttccag aagtttgttg ttctttgctt atgctaccgc aatgtactac 2100

ggcggatttt tgataaggga tggattgcca tacgatagag tgtttaaggt atctcaagca 2160

caaattatgg ggacagtttc aattgccaac tcactggcat tttctccaaa ctttgctaag 2220

ggtgttgcag cagcaaaaaa agttaaaagc ttcctttcaa gaattccact tattcgtgat 2280

ttaccttctt cacgacaaat ggttaaggca agtggcaact tcagtttttc agaaatcgaa 2340

tttacatatc caacacgacc caacgtacta attttaaaag gacttaattt ggacattcta 2400

aatggaaaaa ctgtagctct agtgggagaa agtggttgcg gaaaatctac gataattcaa 2460

ctcatcgaga gattttacga tcctagatct ggtgaagtaa aaatggatgg tgtagattta 2520

aaagatatat cgctagattc gttgaggtcg catatgggta tcgtatcaca agaaccaaat 2580

ctgttcaata agtctattgc tgaaaatatt gcttatggag ataattcaag ggaggtttct 2640

atggatgaga taattaaagc tgctaaaaat gccaatattc ataattttat aactggatta 2700

ccgaagggat atgaaacaaa actaggagaa aaggctgttc agttgtctgg tggacaaaaa 2760

caaagaattg ctattgccag ggcgttagta agaaatccaa aagttttact ccttgatgaa 2820

gccacatctg cactggacac tgaaagtgag aaggttgttc aagaagctct agatcaagct 2880

aaaaaaggaa ggacatgtgt tacgattgcc cacagattaa caaccataca agatgcagac 2940

ttgatatgcg tagtagcaaa tggtgtaatt gctgaatctg gaagccatca agaacttctg 3000

caaaaggagg gactgtacta caaattatat actcaaaaaa cgtaa 3045

<210> 4

<211> 1341

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 4

atgacagaag aaaaaaaaca tagtataaag gataaagaga aaattggtat tgatgcccaa 60

tttgttaata gtgaagaacc aaaggaaaaa attaagaatg tatcttttcc tcagatgttt 120

aggtatgcaa gtacttatga taaattttta atggtagtag gacttatatc ggcaactgga 180

acaggagtct tacaacccct aaatacgatt ctctttggta gcctcacagg agatatcatt 240

gcatatgcta cgtcgataca gataaattta cctgccgatc agaagaaaat agcagaagac 300

aatttttttg atggtatccg atattttgcc ctaatgaatt ctctgatcgg aataggaatg 360

tttgtgttca gttacctagc aacagtcact tttaactatt ctgctatgag acagatattc 420

aaaataaggt cagcatattt gaaatccatt ttaaaccaag atgtaggttg gtacgatatt 480

aatcagactg gagattttgc cagtaggatg tcagatgatt tgttcaaatt tgaagatggc 540

ataggtgaaa aagtaccggt attctggagt tttcaagttg tttttttaac ttcacttatc 600

atagctcttg tgaaaggatg ggaacttgct ttaatatgtt taacctctct acccgctaca 660

ttaataacaa tcggaattgt aggacttctc actacaaaat tagcaaaaaa tgaactggaa 720

gcatatggcg ctgctggatc gatagccgaa gaagcattgt ctttaattag aaccataacg 780

gcttttggag gtcagaagaa agaagtagac agatataata aaaatttagt tgaagccaaa 840

aacaataata taagaagatc gatgttttca gcattaggtt tcggtttatt gtggttcatg 900

atctatgcca gttatgctct ggcattctgg tatggtgtaa aattagtttt aagggacagg 960

acagctacga atcaaatata tactcctagt aatatggtga ctggatatga aacaaaacta 1020

ggagaaaagg ctgttcagtt gtctggtgga caaaaacaaa gaattgctat tgccagggcg 1080

ttagtaagaa atccaaaagt tttactcctt gatgaagcca catctgcact ggacactgaa 1140

agtgagaagg ttgttcaaga agctctagat caagctaaaa aaggaaggac atgtgttacg 1200

attgcccaca gattaacaac catacaagat gcagacttga tatgcgtagt agcaaatggt 1260

gtaattgctg aatctggaag ccatcaagaa cttctgcaaa aggagggact gtactacaaa 1320

ttatatactc aaaaaacgta a 1341

<210> 5

<211> 3771

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 5

atgacagaag aaaaaaaaca tagtataaag gataaagaga aaattggtat tgatgcccaa 60

tttgttaata gtgaagaacc aaaggaaaaa attaagaatg tatcttttcc tcagatgttt 120

aggtatgcaa gtacttatga taaattttta atggtagtag gacttatatc ggcaactgga 180

acaggagtct tacaacccct aaatacgatt ctctttggta gcctcacagg agatatcatt 240

gcatatgcta cgtcgataca gataaattta cctgccgatc agaagaaaat agcagaagac 300

aatttttttg atggtatccg atattttgcc ctaatgaatt ctctgatcgg aataggaatg 360

tttgtgttca gttacctagc aacagtcact tttaactatt ctgctatgag acagatattc 420

aaaataaggt cagcatattt gaaatccatt ttaaaccaag atgtaggttg gtacgatatt 480

aatcagactg gagattttgc cagtaggatg tcagatgatt tgttcaaatt tgaagatggc 540

ataggtgaaa aagtaccggt attctggagt tttcaagttg tttttttaac ttcacttatc 600

atagctcttg tgaaaggatg ggaacttgct ttaatatgtt taacctctct acccgctaca 660

ttaataacaa tcggaattgt aggacttctc actacaaaat tagcaaaaaa tgaactggaa 720

gcatatggcg ctgctggatc gatagccgaa gaagcattgt ctttaattag aaccataacg 780

gcttttggag gtcagaagaa agaagtagac agatataata aaaatttagt tgaagccaaa 840

aacaataata taagaagatc gatgttttca gcattaggtt tcggtttatt gtggttcatg 900

atctatgcca gttatgctct ggcattctgg tatggtgtaa aattagtttt aagggacagg 960

acagctacga atcaaatata tactcctagt aatatggtga ctgttttctt tagtgtcatg 1020

acaggatcta tgaactttgg tattgcctct ccatatatag aggcatttgg aatctcaaaa 1080

gcggcggcgt caaaaatttt tagtgtaatc gatcacaaac ctacaattaa tttatcaaaa 1140

ggaaatggaa aaacgctaaa cgtactcata ggaaatatac aatttaaaga tgtggctttc 1200

cgttatccat ctaggaaaga tgtaccgata ttaaaaggat taagtttgaa cattaaatct 1260

ggagacactg tggcacttgt aggtagttcg ggctgtggaa aatctacggt tattcagtta 1320

cttcaacggt tatacgatgc ggactctgga gaggtcacca tagacggaaa aaatattaaa 1380

gaatacgatt taacttggtt aagaagccaa ataggagttg taggtcaaga accaattctc 1440

tttggaacat ccattttaga aaatattagg tacggtaaag atggagtaac agaggaagat 1500

gtcattcaag cagctaaaaa agctaatgcg cataacttta ttaaagctct tcctaacgga 1560

tataatacat tggttggaga gaaaggggct caactctcag gtggacaaaa acaaaggata 1620

gctatagcta gagctttggt aagaaatcct actttattac ttttggacga ggctacttca 1680

gcactggata ataccagtga agctaaagta caagcagctc tagatgcggc aagtgttgaa 1740

tgtacaacta taattgtggc acacaggcta tcaacaattc gaggtgcaaa caaaattatc 1800

gtactttctc aaggagtcgt ggtagaagaa ggaacccacg aagaactaat ggaacttaaa 1860

caagagtatt atagattagt aacagcccaa gtcaaatcat cggaacagtt tgaagtcgct 1920

gaaaagaaga aagtggtaag agctattagc ttggcagaaa gtagcacagg ctctgaccat 1980

aacatagagg ctacaaaaga agataatgaa gacgatttta atgaaaataa agacgtttcc 2040

gtttttgaaa tattgaagat gaatgctcca gaatggcctt acattttatt tgctggtctt 2100

ggatcaattg ttgttggttg tggaatgccc gtttttgcag tcttgtttgg tagcattttg 2160

gggacattag caaatggtga tcctgacttt gtaagatctg aaaccaacaa atattgctta 2220

tattttgtgc tgggaggact cataactatg gtatcagtat ttactcagat gtacctgtta 2280

ggaattgctg gtgaaaaaat gacagaaaga gttaggagtc gattattcaa ggcaatgata 2340

taccaggaaa taggcttctt tgacaagaaa acaaatgggg ttggagcgct ttgtgccaaa 2400

ttgtcttctg atgcttctaa tattcaaggg gcaacgggaa ttcgggttgg aaccattctt 2460

caatcaatag ccacattttg cttagcaatt ggtctttcca tgtattatga atggaaattg 2520

ggcttagtta cagcagcttt cacacctgtt attttgatcg ctatgttctt tgaaagaaga 2580

aacaccagag gtggaaacga ttcaagagat tccgcactac aaaaatcaac aaggactgca 2640

gtagaagccg ttgggaatat acgaacagtt gcctccttag gcttagaaga aaagttccag 2700

cagctatatg aatctgagct catgccacat tataaatctt cgttaaaaac agttcactgg 2760

agagcaatcg tgtttggtct ttccagaagt ttgttgttct ttgcttatgc taccgcaatg 2820

tactacggcg gatttttgat aagggatgga ttgccatacg atagagtgtt taaggtatct 2880

caagcacaaa ttatggggac agtttcaatt gccaactcac tggcattttc tccaaacttt 2940

gctaagggtg ttgcagcagc aaaaaaagtt aaaagcttcc tttcaagaat tccacttatt 3000

cgtgatttac cttcttcacg acaaatggtt aaggcaagtg gcaacttcag tttttcagaa 3060

atcgaattta catatccaac acgacccaac gtactaattt taaaaggact taatttggac 3120

attctaaatg gaaaaactgt agctctagtg ggagaaagtg gttgcggaaa atctacgata 3180

attcaactca tcgagagatt ttacgatcct agatctggtg aagtaaaaat ggatggtgta 3240

gatttaaaag atatatcgct agattcgttg aggtcgcata tgggtatcgt atcacaagaa 3300

ccaaatctgt tcaataagtc tattgctgaa aatattgctt atggagataa ttcaagggag 3360

gtttctatgg atgagataat taaagctgct aaaaatgcca atattcataa ttttataact 3420

ggattaccga agggatatga aacaaaacta ggagaaaagg ctgttcagtt gtctggtgga 3480

caaaaacaaa gaattgctat tgccagggcg ttagtaagaa atccaaaagt tttactcctt 3540

gatgaagcca catctgcact ggacactgaa agtgagaagg ttgttcaaga agctctagat 3600

caagctaaaa aaggaaggac atgtgttacg attgcccaca gattaacaac catacaagat 3660

gcagacttga tatgcgtagt agcaaatggt gtaattgctg aatctggaag ccatcaagaa 3720

cttctgcaaa aggagggact gtactacaaa ttatatactc aaaaaacgta a 3771

<210> 6

<211> 3771

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 6

atgacagaag aaaaaaaaca tagtataaag gataaagaga aaattggtat tgatgcccaa 60

tttgttaata gtgaagaacc aaaggaaaaa attaagaatg tatcttttcc tcagatgttt 120

aggtatgcaa gtacttatga taaattttta atggtagtag gacttatatc ggcaactgga 180

acaggagtct tacaacccct aaatacgatt ctctttggta gcctcacagg agatatcatt 240

gcatatgcta cgtcgataca gataaattta cctgccgatc agaagaaaat agcagaagac 300

aatttttttg atggtatccg atattttgcc ctaatgaatt ctctgatcgg aataggaatg 360

tttgtgttca gttacctagc aacagtcact tttaactatt ctgctatgag acagatattc 420

aaaataaggt cagcatattt gaaatccatt ttaaaccaag atgtaggttg gtacgatatt 480

aatcagactg gagattttgc cagtaggatg tcagatgatt tgttcaaatt tgaagatggc 540

ataggtgaaa aagtaccggt attctggagt tttcaagttg tttttttaac ttcacttatc 600

atagctcttg tgaaaggatg ggaacttgct ttaatatgtt taacctctct acccgctaca 660

ttaataacaa tcggaattgt aggacttctc actacaaaat tagcaaaaaa tgaactggaa 720

gcatatggcg ctgctggatc aatagccgaa gaagcattgt ctttaattag aaccataacg 780

gcttttggag gtcagaagaa agaagtagac agatataata aaaatttagt tgaagccaaa 840

aacaataata taagaagatc gatgttttca gcattaggtt tcggtttatt gtggttcatg 900

atctatgcca gttatgctct ggcattctgg tatggtgtaa aattagtttt aagggacagg 960

acagctacga atcaaatata tactcctagt aatatggtga ctgttttctt tagtgtcatg 1020

acaggatcta tgaactttgg tattgcctct ccatatatag aggcatttgg aatctcaaaa 1080

gcggcggcgt caaaaatttt tagtgtaatc gatcacaaac ctacaattaa tttatcaaaa 1140

ggaaatggaa aaacgctaaa cgtactcata ggaaatatac aatttaaaga tgtggctttc 1200

cgttatccat ctaggaaaga tgtaccgata ttaaaaggat taagtttgaa cattaaatct 1260

ggagacactg tggcacttgt aggtagttcg ggctgtggaa aatctacggt tattcagtta 1320

cttcaacggt tatacgatgc ggactctgga gaggtcacca tagacggaaa aaatattaaa 1380

gaatacgatt taacttggtt aagaagccaa ataggagttg taggtcaaga accaattctc 1440

tttggaacat ccattttaga aaatattagg tacggtaaag atggagtaac agaggaagat 1500

ttcattcaag cagctaaaaa agctaatgcg cataacttta ttaaagctct tcctaacgga 1560

tataatacat tggttggaga gaaaggggct caactctcag gtggacaaaa acaaaggata 1620

gctatagcta gagctttggt aagaaatcct actttattac ttttggacga ggctacttca 1680

gcactggata ataccagtga agctaaagta caagcagctc tagatgcggc aagtgttgaa 1740

tgtacaacta taattgtggc acacaggcta tcaacaattc gaggtgcaaa caaaattatc 1800

gtactttctc aaggagtcgt ggtagaagaa ggaacccacg aagaactaat ggaacttaaa 1860

caagagtatt atagattagt aacagcccaa gtcaaatcat cggaacagtt tgaagtcgct 1920

gaaaagaaga aagtggtaag agctattagc ttggcagaaa gtagcacagg ctctgaccat 1980

aacatagagg ctacaaaaga agataatgaa gacgatttta atgaaaataa agacgtttcc 2040

gtttttgaaa tattgaagat gaatgctcca gaatggcctt acattttatt tgctggtctt 2100

ggatcaattg ttgttggttg tggaatgccc gtttttgcag tcttgtttgg tagcattttg 2160

gggacattag caaatggtga tcctgacttt gtaagatctg aaaccaacaa atattgctta 2220

tattttgtgc tgggaggact cataactatg gtatcagtat ttactcagat gtacctgtta 2280

ggaattgctg gtgaaaaaat gacagaaaga gttaggagtc gattattcaa ggcaatgata 2340

taccaggaaa taggcttctt tgacaagaaa acaaatgggg ttggagcgct ttgtgccaaa 2400

ttgtcttctg atgcttctaa tattcaaggg gcaacaggaa ttcgggttgg aaccattctt 2460

caatcaatag ccacattttg cttagcaatt ggtctttcca tgtattatga atggaaattg 2520

ggtttagtta cagcagcttt cacacctgtt attttgatcg ctatgttctt tgaaagaaga 2580

aacaccagag gtggaaacga ttcaagagat tccgcactac aaaaatcaac aaggactgca 2640

gtagaagccg ttgggaatat acgaacggtt gcctccttgg gcttagaaga aaagttccag 2700

caactatatg aatctgagct catgccacat tataaatctt cgttaaaaac agttcactgg 2760

agagcaatcg tgtttggtct ttccagaagt ttgttgttct ttgcttatgc tactgcaatg 2820

tattacggcg gatttttgat aagggatgga ttgccatacg atagagtatt taaggtatct 2880

caagcacaaa ttatgggtac agtttcaatt gccaactcac tggcattttc tccaaacttt 2940

gctaagggtg ttgcagcagc aaaaaaagtt aaaagcttcc tttcaagaat tccacttatt 3000

cgtgatttac cttcttcacg acaaatggtt aaggcaagtg gcaacttcag tttttcagaa 3060

atcgaattta cgtatccaac acgacccaac gtactaattt taaaaggact taatttggac 3120

attctaaatg gaaaaactgt agctctagtg ggagaaagtg gttgcggaaa atctacgata 3180

attcaactca tcgagagatt ttacgatcct agatctggtg aagtaaaaat ggatggtgta 3240

gatttaaaag atatatcact agattcgttg aggtcgcata tgggtatcgt atcacaagaa 3300

ccaaatctgt tcaataagtc tattgctgaa aatattgctt atggagataa ttcaagggag 3360

gtttctatgg atgagataat taaagctgct aaaaatgcca atattcataa ttttataact 3420

ggattaccga agggatatga aacaaaacta ggagaaaagg ctgttcagtt gtctggtgga 3480

caaaaacaaa gaattgctat tgccagagcg ttagtaagaa atccaaaagt tttactcctt 3540

gatgaagcca catctgcact agacactgaa agtgagaagg ttgttcaaga agctctagat 3600

caagctaaaa aaggaaggac atgtgttacg attgcccaca gattaacaac catacaagat 3660

gcagacttga tatgcgtagt agcaaatggt gtaattgctg aatctggaag ccatcaagaa 3720

cttctgcaaa aggagggact gtactacaaa ttatatactc aaaaaacgta a 3771

<210> 7

<211> 26

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 7

tcttacaacc cctaaatacg attctc 26

<210> 8

<211> 23

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 8

agggcaaaat atcggatacc atc 23

<210> 9

<211> 28

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 9

tgcaatgata tctcctgtga ggctacca 28

<210> 10

<211> 22

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 10

ggctttccgt tatccatcta gg 22

<210> 11

<211> 20

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 11

accgtagatt ttccacagcc 20

<210> 12

<211> 26

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 12

acaagtgcca cagtgtctcc agattt 26

<210> 13

<211> 23

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 13

ctaactctgg catcgaatac ctc 23

<210> 14

<211> 20

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 14

tgggcgtttc aaggtagatg 20

<210> 15

<211> 28

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 15

ttctccaggt aagtgtaaga atgtgcgg 28

<210> 16

<211> 3481

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 16

atggagacca tggttaatct aaaaagatta acacacatct tcaatacaac agccaagaaa 60

tacaatatga taatatcagc agaaaaaacc aaatgtatga caacatcgaa atacccacta 120

cgatgtaaaa tcgaaattga tgggaaaata ctaaagcagg aagcaaggtt tagatatctg 180

ggaatagata taaccagtta cggagatgtt gaacaggaag tacgacaaca aagcttaaaa 240

gcaagtaaag gggagggatc tcttaatgac acaatttgga agaacgaaca ccaaagacaa 300

gacacaaaag cagcaattag acctatattg acatacacgg cgaagacaag acctgacaca 360

tctaaaacga gacgactact agaaacaaca gagatgaaaa tactccgacg aatatcaggg 420

aaaagtctgt tggataggga gataagcgaa aacataagaa gatcatgcaa tgtagaagac 480

ataaatggat gggtgacaaa acggaaacag gagtggaacg accacattag tagaatggta 540

gaggatagga tagtctctct agtcactttc gctacatacg tattggtgga tgaaaaacat 600

gtattagacg caaataaagc ctatgtttct atctctctat tcaatatcat acgatttcca 660

ctcagcatgt tgcccatgat gttgtcaaat ttagtacaaa cttgggtgtc agttaaaaga 720

ataaacaaat tcatgaatgc tgaagaactt gatcctgata atgttctgca cgaaccctct 780

gaagagaaga ggagagccac ttacgatcga aacggcacat tctcctgggg cgaaggacca 840

atcttgaagg acatcaacat taacatcaag aaaaaaactc taacggccat cgtcggaaca 900

gtcggttcag gaaaatccag tttgatctca gcgttcctag gagaaatgga caaactctca 960

ggccgtgtga actcatatgg aaaagtcgct tacgtctccc agcaagcttg gatccagaac 1020

gccacccttc gagataacat cctcttcgga aaaccctacg acaaaacgct gtatgacgca 1080

gtggtggaag cttgtgccct gaatccagat ctggagatgt tgccggctgg tgatcagacg 1140

gaaatcggcg aaaaaggtat caatttatct ggaggtcaaa aacaacgtgt tagcgtcgcc 1200

agagcagtct actcgaatgc tgatatatac tttttggatg atccgttgag tgctgtagat 1260

agtcacgttg gaaaacacat attcgaaaag gtgatcggtc cccatggatt gctaaaaagc 1320

aaaaccaaaa tcctcgtcac acacggaatc acgtatttgc cacaaactga aaaaatcatc 1380

gttctgaaag acggtagaat ttccgagacg ggaacatatc aggaactcct ggatagaaaa 1440

ggagcattcg ccgaattcct tctccaacat attacagaag aagccgaaac cgaagcaggt 1500

tctaaagtaa aatctaacga actcgatgaa ctcaaagacc aactcgctga cacaccatta 1560

tctcaagaag tagcaagaca actagtcagg cacaggtctc gcgtctcaga atctcaatca 1620

gaaacaggct ccgaccacat cggcaacgga tctatccaac gcctcaacag cgtggacaaa 1680

agcaaccaca aactatccat agatgacggt aagaaaggtc caaagaaagg cgagaagcta 1740

atagaaaccg agaaagcaga aacgggcaac gtcagctggg ccgtctacaa gcattacttg 1800

aagtctatcg gattggtgtt tatgttggcc accttgattt ttaatttagt gtaccaaggt 1860

ttcagtgtcg gctccaacgt ttggttgggg ctttggtcag atgatcaaaa gatcgtcgtc 1920

aacgataccg ttgacactgc aagacgagat atgtatctcg gggtgtatgg tgctctaggt 1980

cttggacaag ttgtaacgat attgtttgcc tcgttggcgc tgtacgtggg atctttaaac 2040

tcagcaagag ctttgcataa cttgatactc ggtaacgtac taaaagctcc atgtactaca 2100

ttcttcgacg tgactccttt gggaagaatt ctgaatagat tcagcaaaga tatcgacacg 2160

ttggacaacg tgttgcccat gacaatgaga ggctggatca cttgtttcta ctcggtcgta 2220

ggtactctag ctgtaatcag ctattccaca ccagtgttta ttttagtgat aatcccctta 2280

tgtttcattt actacttcat gcaaaggttc tacgtggcta cctctagaca agtaaagagg 2340

ttagaatctg tgtcacgttc cccgatttat tcgcactttg gagaaagtgt ctctggagca 2400

catgctatca gagcgtacaa tcaacaggaa agatttattg tggaatcgga aaacaaagtg 2460

gatatgaatc aaatatgtta ttatcccgga attatatcga atagatggtt ggcagtcagg 2520

ctcgaaatga ttggaaactt aattatattt ttcgctgctc tgttcgctgt tctcaagaaa 2580

gatcaggcgc caggattggt cggtctttct gtgacctatt cgctacagat tacacaaaca 2640

ctcaactggc tggtcagaat gacgtctgat gtggaaacca acatcgttgc cgtcgaaaga 2700

attaaagagt acggagaagc cccgcaggaa gccgcttggg aaatccctag caagcagacg 2760

tccccagcct ggccggaaga aggtacagtc caattcaaaa actattcggt cagatataga 2820

cccggtttgg atctcgtgtt gcacgatctg gacttcgaaa tcaagggtgg agagaaggtt 2880

ggtattgtag gaagaactgg agcaggaaaa tcgagtctaa cgttggcttt attcagaata 2940

atcgaagctg cgcaaggaaa catcctaatc gatggaatcc gcatcgacga acttggttta 3000

cacactcttc gttctagact tactattatc cctcaagatg ccgtactgtt ttctggaacc 3060

ctacgtatca atttggaccc attcaacaaa cacagcgatg aagacgtatg gaggtccttg 3120

gaacatgcgc atcttaaaga attcgtgaaa ggactaccag ctggccttaa tcacgaagtc 3180

accgaaggcg gtgaaaatct gtcagtcggc caaagacagc taatctgcct cgccagagct 3240

ctactccgaa aaacgaaaat tctcatcttg gacgaagcca cagcagcagt agatttagaa 3300

acggacgatc tcattcaaaa gacaatcaga acggagttca aagaatgcac agtacttacc 3360

atcgcacata gattaaacac gatcatggac tcagacagag tcatcgtgct agataaaggc 3420

aggatagcag aattcgacag ccctgcgcga ctgctagcca atgagaattc aatatttgcc 3480

a 3481

<210> 17

<211> 2579

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 17

atggaaaacc ttctctggtt acaccatcca aggcctctaa aaatgcaagc catacggatt 60

ctgagactat atgaagatga gggaatttta caatttataa ttcacatccc atctgctcag 120

gcggataata ctcggtgtac cgtcacggat tgcaacgaac gaaatggcag ggatgcgcta 180

gtgacatctg ctaggagtat tatcctagtt cgttcagaga acgaagagaa gaagaaccta 240

gagaagagca tgtgttttgt ttctcgagga aagcttcaaa gatactgtat cataaccaga 300

aatgtagtaa gtctcctact gatgttctat ccaataatgc acgcatacaa tactattatg 360

acgaaaactc cgaacaaaga aaacgcctcc tttctggttt gcgcagtcca agccatttct 420

tggtgttccc atttcctcta cactgtagga ctaagatcga ggttaggtaa aagtcaaaga 480

ggtcctactg caatgggtat cgtctggtgt atggttttcg cgatgaccat cgtatcgttg 540

agaagtgcct acttggaaaa ttcgcaaaga ccaacagaga ataccaaaat ggagctaggt 600

attactttgt attacaccgt cttgcagatc atttatgccc ttagtttgat accaggcggt 660

agcaccacca cattaaattt tcctgagagg tatacagaga tcacagagcg ccagcccttg 720

ataaacgcca atgcctacgg cagattcagt gaagagggag atccaaacga tctgggggtt 780

gctatggaag acactaactg gttgtcccgt ttatcgtttt cctgggtaaa gtctctagta 840

aacaaaggag ttgaagataa aatagttaca tctgacgatt tgtacgatct accggactca 900

ataagtagcg acaacaatag taatagacta gaaaattacc tgaggattga acacgctgct 960

ctctgtgatt cccactttaa tttcaagatg tctgtgatcg gtctcaaaat gaggggagct 1020

ataatcaata ctatatacag gaaaacttta tcagtgagat ctacggtttt aatgtctaaa 1080

ttatctgtgg gggagataat gaactacatg tctacagata cagaccggat tgttaattct 1140

tgtccaagtt tccacgcagt atggagtata ccattccagc tatttgtatc cctttacctt 1200

ctgtacaacc aagtgggctt ggcgttttta gctggtgtcc tgttcagcat aatcctcatt 1260

cccataaata aatgcatagc gaacaaaatc ggcgatctta gtaccaagat gatggaacaa 1320

aaggacggca gagtgaagtt gacttccgaa atactcaggg gcatcaaggc aatcaaattg 1380

tacgtttggg aacaacactt tataagactt attactagca aacctgtcga tgtactccgc 1440

aacatagacg acactttacc cgtagattta gaactggagg atagtattca atctgaatat 1500

acatcgtctg agtgcactct agagaatagt ttgtgcgtgg acaacgccca agaaaaggat 1560

aaggatatcg aactgttcaa agaagtcagc gaacggggta atctagattt tactgtgata 1620

atttcttact ggaaaggcat cagccacttt gtcagcattt caatactgat ttcgatgact 1680

ttaatgcaag tgtcacggaa tttcactgac tggtggttag ccaatggtgt aaccagcccc 1740

gttaccaatt ctaccaactt aacagtatac ttagcgacgg cgactgacta tatcgattac 1800

gtagaagacg atgataacat gtcagagttt ttaaaagtat acgtcgagtt ggcgtgtgtc 1860

aatacgctat tcacgctaat aagatcgttt atatttgctt atggtggaat tctggcagct 1920

acaaagtttc ataaggtgtt attgaagacc gtgttacgag ctagatgtac atttttcgat 1980

gtaacaccaa taggaagaat cattaatagg ttttctagcg atacctacac tgttgatgat 2040

tcgttgccgt ttattcttaa tatatttctt gcgcagttat tcggattact aggttctctt 2100

tttataacga tgtacggttt gccttggata tgtgtcttcc tcattccgtt agtacccata 2160

tatacatatt tattaaatca gtacaggata acgtcaaggg aactaaaaag gatatctagt 2220

gtaacgcttt ctcccatata taaccatttt aatgaaactt tgcaagggtt gactacaatt 2280

agagcgatga gagtgatcca caaattcaaa cacgacaaca atcaacatct agaagctaat 2340

ctgaaagctc aattcgccag tcaagcggca gctcgatggc ttggtcttag attacagttt 2400

ataggagtga tcataattac aggtgttagt tttattgccg tcattcagca tcagtatgat 2460

atcgcggatc caggttttat cggattggcc atttcgtacg ctctgggaat cactggatca 2520

ctcagcggcg tcgtcaactc gttcacggag acagagaggg aaatggtggc cgtcgaacg 2579

<210> 18

<211> 2488

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 18

aatatgctga acatatcata ctgcccacca aatatttcag tatgggatat atgggtggat 60

catggaatcc cacagtgttt catgaacact gtgacttctt ctgtattagc aatatatata 120

ctaatagctg gttcaataca actacatatt tacagacgat ggggtgttga atctggaacc 180

aggtttttgc ctagagggag attatattat ttacaaacct ttttaatatt gtttatacca 240

gtgttagaaa tagttagatt tattttaacc gcaactgttt atgatgataa acatatttat 300

ggatatatga ttgtttctct agtattaacg acatttgctt ttccgttttc tttatgggtt 360

ttaaaagtgg agttatataa cattcttcca tcagttcaaa caagaggcca tgggatagtt 420

ttattgttat tttggacatt ggcctttata tcagaaaatc tagcgtttat caatttgacc 480

caggagaact ggtggtttaa actgaaggat ttgaccgatc agctggaaat ggcattattt 540

attttacgat atattgcttg cctacttatc tttttcctgg gcctcaaagc tcccggaata 600

gtaaacgaag ttgattatta tatgttaggt ggtcatcaaa ggaatgttat caatccgaac 660

aatgaaaatg cctctacatg gaagaatttt tggaaaaagg tgaagatatt agcgcctttc 720

ttatggccaa agaaaaattt cacattacag tttaaagtac tgatttgctt catattatta 780

ttaggaggaa gggcggttaa tttgtttgtg cctatctatc aaaaattaat tgttgatagc 840

atggaagaaa ctgtggcaaa gatgcttttc agatgggact gggttctcat atatgttggt 900

ttaaaatttc ttcaaggagg tggcactggt ggcatgggtc tattaaataa tgtgagatcg 960

ttcttatgga ttagagtcca acagtacaca acaagggaag ttgaagtaga attatttagg 1020

cacttacata gtctttcgtt gaagtggcat ctaggaagaa aaacggggga agtgttgaga 1080

gtgatggaca gggggacaga tagcatcaat aacttattga attacattat attttccatt 1140

tttccaacta tagtggatat aatagtagca attgtcttct tcgtttctgc ctttaatata 1200

tggtttggac tgatcgtttt tactacaatg atactttaca tagttctaac gatcgttatt 1260

accgaatgga gaacaaaatt tcagcgtcgt atgaatttag cagataatga aactcgaagc 1320

agaagtgtag attctcttct caattttgaa acagttaaat attacggagc tgagaattac 1380

gaagtggatg cttttaggga agctgttctt aaattccaag atgaagaatt taagtcaagt 1440

atcactctta atatcttaaa cacagtacaa aatgttataa tatgcggagg gctattagcg 1500

ggaagtttgc tctgtgttta tatggtagta gaacctaaaa ctctaaaggc aggcgattat 1560

gtattatttg ctacatatat agtacaactt tacgtgccgc ttaattggtt tggtacctac 1620

tacagggcca ttcaaaagaa ttttgtagat atggaaaaca tgtttgacct tctcagagaa 1680

gaacaagaaa ttattgacgc tccaggagct actaatctat ctgttccaag gggccatgtt 1740

gagtttaaaa acgtcagttt tggatatctt cctgagaagt tgatattaaa aaatataacg 1800

ttcagtgttc cttctggaaa aactgttgca ctagtaggac catcaggaag tggcaagagt 1860

actattataa gactcctctt caggttttat gatgtcgata caggagttat tgttatagat 1920

gggcaaaata ttaagactgt tactcaagaa tcactaagaa gagctatcgg cgtagttcca 1980

caggatacag tattatttaa taatactgta gaatacaata tacaatatgg tagactaaca 2040

gcaacagtga ctgatgtgat agaagccgct agaggtgctg atatacacga aaaaattctt 2100

acctttccag aagcttatga aaccaaggtt ggagagagag gactaagact aagtggtggt 2160

gagaagcaga gagtggctat agctagaacg ctattgaagg ctccaaatat aatcttactt 2220

gatgaagcta ccagtgcctt agatactcaa acggaaagaa atatacaaga atctctgaat 2280

agaatgtgtg caaacaaaac cacgataata gtagcacata gactctccac cattattcat 2340

gcagatgaga tattagtttt acaagaagga gaaatagtgg aaagaggaaa acacgaccac 2400

ctgataggcc aagaaggaat atacgctaac atgtggcgac agcaattgga gaataaagat 2460

aaagagtctt tggaaaatag ctcggaga 2488

<210> 19

<211> 4929

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 19

atggccgacg acggcaaatc actttttttc tcgcaactga aagcaatgct caaaaggaat 60

ttattactaa aaaagaggca aaaattaaaa acgataggtg aaatcttgtt ccccatctac 120

tttctagccc ttctggttgt tatcgagttg gttctacctg atccaaattt accagaaatc 180

agtactccaa gaggagaaga atatttattt aaatcttttg acaacggtga atgtcataaa 240

atagcatatg caccaaatac aacaacaaat atcgactttg tcaacaatgt aaccgaaaca 300

tggaaaaaga tgtccaattc caaatcgagt ctagaatggg tcccctatag tacagaggat 360

aaggtccagg aagcttatga tcaagatcat aaatttgcac cgatggcagt gatctttaag 420

gttccttatg tgcatacctc attaagttac cagattcgta ctaatcccta ctatatcagt 480

acccctagtt ctagtgatat tggctgggat aaacagtctt gtaggggtgg tacaaaccca 540

aattcaaaca tggaagacgg cagtacatgt ccagctaata cttactactt ttctggattt 600

ttggctcttc aaaccctatt agactatacg cgaatacgac ttgataatcc aactcaagat 660

aatataccgt atatatcttt ggaaatattc ccaaaagctg caactactgt tggtgatttt 720

gcagacatga ttcgcattat aattccaatt tatatggtac tagctctttc tcaattcgtt 780

acgtatctaa taattttaat agttggagaa aaggagaaaa agattaagga aggaatgaaa 840

cttatgggat tgatggacaa agtgttctgg ttatcatggt tgattatata cgcatgtttt 900

gtaattcttt tggctacaat atgcgttttg ctgttatttg ccttcaaggt atttgttcat 960

gtaaaccttc tactaatctt catcttaatg gtcctctatg gtatttccct cattatgttt 1020

gcatttatgt taacaccatt tttcgacaaa gcgaggacgg ctggagtact aagttccttt 1080

atattggtac taataattgt tttatatttc gtcgagcaaa ttgtctccgg taaacatcct 1140

caatatttat gggcactgtc ccttttaagt ccggctggat ttgcatcagc tcttgataaa 1200

gtagcaatat cagatattca aggaaaagga ttagacttca gtaatatgtg ggataaccat 1260

ggttcggggg ttccttttgg tggcagtttg gttatgataa tggtcgatat agtattgtat 1320

gccataatag gctggtatct tgataatgtc attccaagct catatggtgt aaaaagatcg 1380

ccaatatttt tcctgatgcc atcttattgg agatccagtc ataatcagat ccctcccaac 1440

caacttaata cagaaagttc tgtagatgtg gagccggtgc caagggaaat gcaagacaaa 1500

gaagcaatca gaattataaa cttgaataaa agttttacgc aatgccgtaa accaaccgtt 1560

actgcgctcg atggtatcaa tttgtcgatt tacaaaggcc aaatcactgc aattctgggt 1620

cataatggtg ccggaaagac tacgctcttt aatattttga ctggattgtc ctcgcctact 1680

agtgggagtg ctttggtttt tggatatgat gtcagtaacc ccaatgatat ggacaaaata 1740

agaagaatga caggagtttg cccgcagcat gatatcctgt tcgacgatct gacaccaaga 1800

gaacacttgg aattctttgc agccataaaa ggaatatcga atagacagtc cgcaatcgag 1860

aagataataa gagaaataga tttacttgac aaaattgaca cagcaagtag aagcttaagc 1920

ggtggacaaa aacggaaact ctctattggt attgctctca ttggtgatcc aaaaatcatt 1980

atacttgatg aaccaacggc tggcgttgat ccctattcaa gaagacattt atggaatgtt 2040

ttgcaaaatg taagacgcga caaagtgatt ttattgacta cacattttat ggacgaagct 2100

gatattttag ctgatagaaa agctgttgtt tccaaaggaa aaataagatg ttgcggaagc 2160

tcattatttt tgaaaaacaa atttggaatc ggctaccatt tgacatttgt cctggaggat 2220

aagtcaaatg aaaatgctat caaccatttg gttctgcaat tcgtcaaaat ggcaaggaaa 2280

gatagacgac acggaaaaga attgagtttt atattgccac acaatgctgt tgaaaatttt 2340

gcttcacttt ttgcggccat agaacacgag atcagcatta aatccgattt aggtatttcc 2400

agttatggcg tttcaatgac tactttggaa gaagtttttc ttggtttgca aaaagaagag 2460

gaatatggcg acgcaacggc ataccaggat ttgccccaga gctcagaagg gattagcaac 2520

gaaaacagag ctcttaccga aggtattcgt tcattggaag catttcgctg tactcctagt 2580

acttcgcaaa atttaagaac gttgataggc ctacgacttt tgagactacg acgtgagaaa 2640

aggaagctgt ggatggtaat catcttgcca attattttta caggtctagg actgtactta 2700

aataaggtta ttgacactgg aaaaaaatac gaaccaccta aaacatatcc aatgccatta 2760

gacttgagct attacctagg ctataatatt agtatctaca atgggagtcc ggcagattta 2820

actacattta agaatcagtt agagaaaaat ggagcagagc tagatactta tgatggagaa 2880

tttacctcat tgttagacat tgctccacat tatgcatctt ttaacattca cgattttagt 2940

tcttcttctc aaaatatatc cgtactttac aatggcagct attccaataa cttaccaatg 3000

tttattaatc ttatcagcaa tacttttaca gttgcaatgc gaagtatggc cgcaacaact 3060

ggaaaaatcg ccgttaccac ataccctttt gaactacctc caactaatgt gagtcctact 3120

cctgctagtc cagggaactt cattttgggt atgattttcc tgtttgcgcc aatcgtacta 3180

gcagttgata tggtgtacga aagagagata aaagccagaa atcagttaag agttaatgga 3240

cttcctttca ccgtttactt tacaagcttt ttcctggttc aaatcttatt aatggcaatc 3300

ataactatat tgttaataat tcttattgtt gttatgaaac caccgacttt ttcaaattcc 3360

tcaataacag ttttagcaat atgggttatt ctctattgcc ccgcttcagt tttgttttgt 3420

tcctgcgtta gctacatgtt tgataagtcg gagtctgcgc aatcggttat gccaaacgta 3480

tctactcttc taggattaat accatatata tctgtgatgt atgcaaaaga aagtgttgca 3540

actatattgc attacgtatt tgcatttact gatatgatgt atgttcctta tggtatgctt 3600

tattatatac aaaagattaa tttggaatgt atatctgatc cccattgcga gggaactaca 3660

ttctccagct ttatgatacc ggaaattata gtactatttg ttacgctgat aattcaaata 3720

cctctcttgt tcctgattat gcttattctg gatgttaaga aaaatggagg acaagttttg 3780

gatacattta gatcaaagaa aatttccgaa aatattgttg aagagtccag agatgtcgga 3840

gtacatgagg acagagatgt taaaaatgag agacagcgag taaacaattt aattagggat 3900

cctcaaaata accgctctgt tataacagtt gagaatctac ataaggttta tcaaaaaggt 3960

gttaaatctg gtatttgctc acgttctgcg gaaagtctga aggtcgcaat aaagtctata 4020

tcattggcag tagattcggc agaagtattc ggacttcttg gacacaacgg tgctggaaaa 4080

actacggcaa tgaaaataat tacagctgaa gaggcaccaa cacgaggacg ggtacaaatt 4140

gttggaagaa acataacctc cagcgctaat gcagcattcc agtatttggg ctactgtcca 4200

caacatgatg cccaatggaa aaatataact gtgaaagaac atctagaatt gtattctcac 4260

ataaggggta ttccgaagaa tgaaattaaa agaactgttg acttgtactt aacgggtctt 4320

caaattcacg aacataaaga taaacaggct gaaaaatgtt cgggtggtac cagaaggaag 4380

ctcagttatg ctatggctat gattggcaat ccgaaaatag ttcttctcga tgagcctagt 4440

actggaatgg acccgcaaag taaaagattt ttatgggata ctgttttagc tagttttagg 4500

ggttcaaggg gagctattct tacaacacac tccatggaag aagctgatgc gttatgttcg 4560

agaattggca ttatggtaaa tggagaactt agatgtctag gatcaagtca acatcttaaa 4620

aatctatatg gagctggata caatttagaa gttaaattgg caacacagca gggtacagat 4680

ttacgccaaa aattaaaaga attagaaaac tatgtgttcc atctatttcc caatgcagta 4740

ttacaggaaa cttttggcga caggctcata ttcagcgtgc cacaacaaag tgttccttcc 4800

ctggccaatt gttttaggag tttagaggac ggtaagcgtc gattaaatat tgaagaatac 4860

agctttagcc agacaacatt ggaacaagtt ttcctgaaat tcgcccaaga aggtgaacat 4920

gaagattaa 4929

<210> 20

<211> 4164

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 20

gaactaatga aaatggtagg aatgaaatcc tggatgttat ggtttggctg gtttatttat 60

gccatgatac cgatgttttt cgccgttagt gtgatatcta tttttatgaa agtaccaatg 120

tttgggtctg actctccact tgtagaattt gccaacggcg gaatactatt cgtgtttcta 180

atattgtatt gtatggctgc agtagcattt tgttttgcca tcagttcgtt tttcagccga 240

cccacaattg cattagtcgc aggaattctc gtatggattt tatcattttt cataccaaaa 300

tacgcatgtg gactagacga agcaaacaaa ttgtcttggt tatccaacgt tctgttaaac 360

ctactaccaa atatgtccct tcattacgga tattccatca tttctgtatt tgaagaaaga 420

gaagtcggca tcaattggtc aaatttcttc aagcctggta gtggatcatc agatgatatc 480

accatgttaa acgtttatgt gatgttaata gtcgatattg ttatctacac cattttcaca 540

ttttatatgg atggagttaa tcctggtaaa tacggggtcc gaaagagtat attattccct 600

ttacaaaatt ttatgaagct ctgtagaaaa ccatctatca ccacagtgcc agtagattct 660

gaaacagttc ctttggagaa ggttgaagca ggtcacaatt tagctaaggg tattcaaatt 720

aaccatcttc ataaaaggta taaacaaaaa caagctgtca ataacctgaa tttagacatt 780

tacaaaaatc aaatcactgt actacttggt cacaatggag cagggaagtc aaccaccatg 840

tcaataataa caggactgat tccagctacg tctggttcgg tgacaataaa tggattggat 900

attaatactg acatggacga aatcaggaag agtttgggac tatgtcccca acataatctt 960

ctgtttaccg atttgacagt taaagaacat ctcttatttt tcgccaagct gaaaggaaaa 1020

tctacgaaag aagccaacat agaagcaaaa agtttattgg caaaattaaa tatgcccgat 1080

aaagagcatt cgatggctca gacactctca ggtggtatgc aaagaaagct atgcttggca 1140

atggccctta tcggtgattc aaaggtatta attttggatg aaccttcttc gggaatggat 1200

ccccaatctc gaagagagct ttgggatcta ttgttgcaat ggcgagggga gaaaacaatc 1260

cttattacca cacattttat ggaagaagca gatgcactag gagactggat agcaataatg 1320

tctgacggta gtctaaactg ttatggtact ccaatgtttt tgaaaaagaa atacgataca 1380

ggttaccact taaatttaat gattgaagaa gacgcagata tagatgccat tagtagaaga 1440

gttaaacatt ttatgcccga tgcgcatttg acgtcaaata atggaaacaa cttggtattt 1500

gttttaccct atcagaatac caacatgacg gggcttcttg gagatttgga gaaaaatcga 1560

tccgagttac accttagtaa tatatcgata acgataacga ctttggaaga tgtattccta 1620

aaaacacgcc aagagattga acattcatca gacgacactt catcagtggc cacacaagat 1680

gacatccaac ctcatgcctc tttgactctt attactttac tcaaaaagaa agtgaatttc 1740

tctctaaaaa agtggtcaac atacattgta cctgtgggtg tagctgtagt ccttttttct 1800

ctgacggtat ttttaagttc tggaaatgac tactatagtg aaaggggacc agaactaccc 1860

ctcaaactca gcagctacaa acaaacatcg gtttactaca gcggggacac atcaaacgaa 1920

aaaattaata cgttaatgaa atattacatg tcagctgtag aatcgcaaag aagtttacca 1980

ttcaaagttg ataacgttga aaaatctatt ttgcaaagaa gtctggaaaa catagcttac 2040

tataaagaac atatgattgc tggagctaat ttcgtttatg atcgtgattt tacattagta 2100

gcattataca atggtatagc tactcacagt actccgattt ctttaaatct aattaccgac 2160

tcgctggcta aaacgttatt aggtcccgaa tatggaatat ctgtcagtaa ttggccttta 2220

ccgcaggtcc aagagcgact ctcatcacag gagtattcgg aagcaaaagt tgctgttctg 2280

tggctcatcc tgttaccggt gggatgtctt tttattcacg gcatttttat catattccca 2340

catactgaaa tatcgacgag atttttacaa atacagtaca tggctggcgt gaaaccattc 2400

ttctattggc ttgttaattg gattgccgat atgacatttt acatattttt aatgttcata 2460

ttaagtttct tactgtggat atgttctccc gtattccaac ataacggtac cgaactcggt 2520

tatttatttt ccatatttct gtgttatggt attgctggga taccttttgc gtatatattc 2580

agccgaaaga agacagcatc tggtgccttt gcgttattcg tcatcatggg gatgttcttg 2640

ggcatcattc ttacactaac catagccgta ttactagaat cgcaagatga atactacgta 2700

aatattggaa ataaaattaa atatgtgtgt ttcttcttat taccgcaagt ttcattatca 2760

gatgcattgg tctcgtttat tagacgcaca gttaatattt acaatttcaa aatatctccc 2820

caacgtttga gagttatgtg taatcagatg gagtcacacg cttgctgcgt tgaaagttca 2880

atcgaatgtc aaaattataa gagctacaac aacttattca gcgaacatta catgtttatg 2940

ataggatgcg gagtttttta tctaacgata aacattattc tagacacata ttttatgaag 3000

aaattaaaag ctatggttat gcgtcaatgt aatcttattt tgaaatcatt caaagacaaa 3060

gatacagcac taatccccaa aaatgacaaa tatgttaacg aagatgttgg tgacggctac 3120

aatactttaa gagccaagaa aataatgaaa ttatatgccg gaaaacagat agttaaaaat 3180

ataaatttca ctcttaagca cggtcactgt ttgggtatat taggtgtaaa tggggctggt 3240

aaaactacaa cctttaaaat gcttacaaga gaggaagtgg ttgatgatgg agaaattaag 3300

atcgaattag acaataataa acaacctctg gacatcacag gaagcgagta tttggagaca 3360

ataggttatt gtcctcagtc agattcgcta aactttgtcc ttactggcag acaactacta 3420

agtaccgtgg caaagttacg aggtgttact gatgaaacaa tgattgatcg ctttttggaa 3480

gcttttgact taaaacaata tgcggatatc ccgtgcggac attatagtgg aggaaacaaa 3540

agaaaactaa gcttggccat atcccttata ggtaaccgta agtttgtatt attggacgag 3600

cccaccaacg gcgtagatcc ggctactaga aggaagtgtt gggatctgat taaactcatg 3660

aaaggcaata gtggaaacaa gattggattc attttaacat ctcatagtat gactgaatgc 3720

gaagctttat gcgacgagct gaaaataatg aagaaaggtt cgtttgttga agaaggaaga 3780

ttggtggact taaagaatag atatggtgga tttacattaa aattaaaact tgtgtctaat 3840

caaggtcagc ctataaatct ggtagatgat gacgtagacg aggtagatgg agtaataagt 3900

aacaaattta aaagtgtcga tgatcttaaa tcctatttta cgaatcatga tcgcggtgaa 3960

atcaaagatg aacattcagg tctcctccat gtttacatta aggataaaac taaaaaatgg 4020

agcgatattt tccaagaagt ggaggcactc aaaacacaca attcgcattt aattgaagat 4080

tatgcaatca gtgaagcttc gttagaagac gtctttttga aagttgccag agaagatgaa 4140

gaagatgtta ccaaaaagaa atag 4164

<210> 21

<211> 4089

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 21

atgtggttca cggtaagttg gcactggcat gttaaacaga taggccggat atctgatgac 60

atcaggaagc cgtggtttct gccgtttttt aaatttggat acaaacacga catcacaata 120

aaagatgtat ataataccac acaaggtgat ttatcagccg ctcttgggga tgcgttgcaa 180

aaaaattggg aagatgaaat tagaaatcat gaagacaaaa agcaaaagaa gcctagccta 240

aaaaatgcca tattaaagac attttggaaa tcttattcca gatctggcat agcattaggt 300

tttcaatttg tagttattag aatgcttcag ccgatagtat tagccgaatt cataaactat 360

tttgacagta atcaagaagc ttacattggt tggtggctgg caactggtgt aatcggaatg 420

gcctttctta atgttatcat aacgcatagt tgtactttag atacccaaag ggtaggaatg 480

cgagtgcgta tagcagtgtg ttcgttaatt tatagaaaac tcttgaagtt aagccacaat 540

tccctgggtc aaacagcatc cgggcaactg gttaacctct tatctaatga cgtccaaaga 600

tttgacttag ccgcccaatt tatacattat gcctggatta tgccacttac tgcagggata 660

tcgttttata ttttataccg atatgttggc atcatcgctg ccgttactgg aatggttttc 720

attactctgg agtctcttcc tttacaaggt agtttctcga aatggcaagg aaaactaaga 780

tacaaaattg ctctaaagac agacaaaaga gtaaaactga tgagcgaaat tacttctgga 840

atacaagtca tcaaaatgta tgcatgggaa aaaccattcg aaaaagtcgt agagctctct 900

agaaaatacg aaatagatat tataacaact acctcttatt gctatggcgt cctatcagca 960

atgggcatat tcacagaacg tatgatttta tacatcacag tcattacttt tgtactagtt 1020

ggacaaagat taacgggaga tgtagttttt tctttagctc agttattcaa cactgttcaa 1080

ttgatcatgg cgatattctt cccaagagct ttgtcatttt attctgaagc caaagtttct 1140

attacaaggt tagaagactt tcttttatta gatgaaaacg aaaaaatacc ggagccggaa 1200

aaaatctcca acccagacga attaggagaa attcaattga ctaatatttc agccagttgg 1260

gcatcgaaac caattgcacc cactctgatg gatctcaatt tacacattca accgggaact 1320

ttatgttgcg tcgttggtaa cgtaggctgt ggaaaaagta gtcttctgca gttacttctg 1380

agggaattac cggcaagtag ggggcaaatg aagattaacg ggaaaatttc ctatgcttcc 1440

caagaacctt ggctttttgt ttcaaacgtg aaagaaaata tactttttgg aaaaccattc 1500

ttgaaaaaca ggtaccatga cgtagttaag gtgtgctctc tagaaagaga ttttaaacaa 1560

tttccctttg gtgatagaag cttagtaggt gaaagaggaa catcgttaag tggaggacaa 1620

cgagcaagaa ttaatcttgc cagagcagta tacactgaag cagatattta cttattcgac 1680

gatccgctat cggcggtcga cacaaaggta gcaagacatc tttttgatga atgtatatcg 1740

aaatatttat atggaaaaac tagaattttg gtcacacatc agctccagtt tatgaagaaa 1800

gctgatttga taattattat aaacaatggt caaattgaga aaatagccaa atttaacgaa 1860

ttgtcggaaa atgacttgaa tgctcttcaa caagaacctg aggccgatga taaagagaaa 1920

gaaaaggtac cagagactgg cgataagatt cgtaaaaatt cgacagtacc tcatttccag 1980

tctatgagtt ctttggctag ttctgtattc tcggatgatc cgaacgaaga agacgaactt 2040

atcgaaaaag gtgccatttc aaatgctact tatgtggaat attggaaatc aggcggtgga 2100

gttttattct tattctttac aatttttata ttcctcgtcg ctcaaatgat tacgaatgca 2160

tcagatcttt ggcttacaca ttggactaat aacgaggcaa aaagatacca tcttagtgat 2220

tccctacaca atgcaacttc caataaaacc ttagaaagac cattactaat gtctactgga 2280

ttatctgatg agatgtcaac ttctgctacg ttattaacag attcaacacc atctgcagat 2340

agttcacttc taagtttaag tcagaaacga tatttaacag acctcatcag aaatgctact 2400

ttagaagaac tgaatgacat tccgtctaca gaatattata tttatgtgta cactggaatt 2460

atattagctt ctgttgtatt tcttacgtgg agatcattcc tttattacca gatatgtatg 2520

actgcttcca aagttttgca taataagatg tttaacaatg ttcttcaagc accgatgaga 2580

tttttcgata caaatccatc agggcgaatt ttaaacaggt tctcaaaaga tatgggtgca 2640

gtagacgaac ttttaccacg ctgtcaaatt gacgctattc aaatttttat ggttatgatt 2700

ggtatcttag ctatggtgtt catagtgact ccttggatga tcattccagc agtaatactt 2760

gctcctcttt actacttctt ccgtgtagtt tacttaacaa gcgcacaatc gttaaaaaga 2820

ctggaaggag tgtcaagagc tccggtgttt tcccacatat cggcttcact atacggtata 2880

acaacaatta gagcctcaaa cgctgaaaaa atgataacta cagaatttga tatcctacaa 2940

gatcagcaca ccagcacgtg gtacttattt atagtcagca gtaccgcttt tggattttat 3000

ttggatgtca tgagctgctt tttcttggcg attgtgactt accaatttct tctttttaga 3060

acagaaaata ctttaagtgc aaacgtgggc ttggtcatat ctcaaagtct catattaacg 3120

ggcatggtac aatatggagt gcgacaaagt gcagaagtgg cgagtaatat gattagcgta 3180

gaaagagttt tgcaatatac taaactggat aaggaagggc ccttcgaaac gctacctggc 3240

aagaagccac caaggaattg gcccgaaaaa ggaagaatta tatttaagaa tacgtactta 3300

aaatatgctc cagaacttcc acctgtattg aaagatttaa atatagaagt aaattctggc 3360

gagaagattg gaattgttgg acgaaccggt gcaggaaagt caaccctcat tgcgtcactt 3420

ttcagattag caccgataga gggtactata tcaatagatg atgttgatac tgccgaaata 3480

ggtttgaatg atctaagatt aaatatctca atcatacccc aagaaccaat tttgttctcc 3540

gcttcactca ggtataattt agatcctttt gagaaacatg gtgatgaggt tttatggaaa 3600

gcgttagaag acgttgaact caaaggtgct attagtgact taaatcaagt tgtcagtgaa 3660

ggaggatcta acttcagtgc tggccaaaga caacttatat gtctcgctag agctattata 3720

cgcaataaca aagttctcgt tatggatgaa gctaccgcta acgtagatcc acagactgac 3780

ggcctcatcc aaaaaactat tcgtgagaga ttccaagact gtacagtact tacaattgcc 3840

cacagattaa acaccattat ggattcagac agagtattag tcatggatgc cggacaggca 3900

atggagtttg accatccata tcagcttcta caaaatccgg aaggatactt tagcaagatg 3960

gtccaggaaa ctggacccgc catggcggag ctattaagta atgtagcaaa gaatgactac 4020

attaagaaga agggaccgtt aaccagtgaa gttccacaaa atttagcaat tgaagataac 4080

aaagattag 4089

<210> 22

<211> 3972

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 22

atggaaatga ccaaagagaa atataatccc aatccacggg agaaagcgaa tattttatcg 60

actctatttt ttggatatac atatgatgtg tttaaaaagg gcttatcgaa aactctagaa 120

gttgacgatt tatataatcc attaaagatc gatcgaagca aaatattggg agatcattta 180

caaagcaact gggacaagca actagaaaaa gccaagaaaa caaactcaaa accaagttta 240

ctgaaagcca ttcttatgac atattggtta gagtatttga agttaggaat attcgacata 300

acaacagatt tagtccttag actcatccaa ccattaatgc ttggaaagct gttggaccat 360

tttaaacctg aagcacaagt ttcaaaaaat gaggcactca tgtatgcagg aatcatagtg 420

gctataaacg tattgaactt tttgatcgga aatcagtata tggtagaggg tttccacagt 480

ggcatgagga taagagcagc ttgttgtgct gttatatata gaaagtcctt aaaattgagc 540

aaaacagcct tgggagaaac agcttcaggg aaactggtca acctcctatc caacgatgtc 600

agtcgatttg atctcgtcag tttactcatt catcaaatat ggattgcacc tgtattgtca 660

cttatagtaa tggttttatt ataccagaga actggatatg ctggaatagt tggggttgtg 720

gcagtttttg tgatagttcc tattcaaaca tacactggaa agctatctgc aaaatacaga 780

aaacagacag ctatgaaaac agacgagaga gtcaggctaa tggacgaaat catctcagga 840

attcaagtaa tcaaaatgta cgcatgggaa ataccgttca ggaaggtcat acggattgct 900

agacgaaatg aaatcaaaat cattacaaaa tctgcgtatg tgagagcttc atttatggca 960

ttgaacctgt tcacaactag attggcgtta ttctgtactt tgcttactgt tattttgtcg 1020

aacgaaccaa tcacagcctc aatggttttc gttgtaatgt cttacttcaa cgttatttca 1080

atggctatgt ccaccatgtt tactagagga gtatcagaaa tagcagaatg tttggtagct 1140

gttcggagaa tcagagattt ccttctaaac gaagagtacg atcccaatag atcaacttct 1200

ggtatgaacg gcaatgtcaa aagtgtagac gattttaagg aaattatcag tttgaaaaat 1260

ttaactgtaa aatggaattt aagttttagc gacaatgctc tcgagaatat aaatttaaat 1320

gttcaagatg gacagttgat aggaattatt ggacctgtag gtagtggaaa gagttcttta 1380

ttgcaaacct tactaggtga actggacatc acccaaggcg aaatgagagt acaaggtcaa 1440

atctcctatg cctcccagga accgtgggta tttgcagcca ctgttcgtca aaacattctt 1500

tttggtgaag aatatgataa aaagagatac caggaagtta tacaagcttg ttctttagaa 1560

aaagattttg aacagtttcc aaatggagat ttaactctcg taggtgatag aggatcatct 1620

ttgagtggtg gtcagaaggc taggattaat ttagcaagag cagtgtatag agaagcagat 1680

gtctatctct tggatgatcc gttatccgct gtagatatac atgtttccaa acatttatat 1740

gaaaaatgta ttaatggcta tttggcaagc aggacgagaa ttttagttac ccatcaagta 1800

cactatctga aggatgctga caacatcatt attttaaata atggtcgcat cgaagatgaa 1860

ggaactttca actttttggc aaatagtgac aatgtgtacg caaaactctt aacagctgaa 1920

ccagaaaata aggaagagaa gaaaccagag agacagaaat attctaggca actatcacaa 1980

agaagtcgaa aagattcaat gtcaagtata ataagtgaat tgagtattgc ggatactctt 2040

ttatcaaacg atgtcgactt tgacgaagag gaagctgaaa aagaaccaga gtttgatgtc 2100

aaagaccttc aggaacaatc atcgaaggga aaagtgggcg gttctttgtt atttaaatac 2160

atgttagcag gttctaatgt atttgctgtt ttcatatgcg ttctactata tctcggtact 2220

cagttagctg ccagtgggac ggattactgg gtaagttact gggtgaatgt agaggaattc 2280

agaaactcgt ctgaatcttt gaattctact gcacctaggt tcatcatccc atccatcgaa 2340

ttaacaacag ataattgcct ttatatatat tcagttatct taggagctct ttttgtcctt 2400

gctatgacaa ggtctttctt cttttataaa atggctatgt tgagctcgaa aaaacttcat 2460

ggcacaattt tcgataatgt tattgatgct acaatgagat tcttcgacac aaatcctgga 2520

ggaagaattc ttaacaggtt ttctaaggat atgggagctg tcgatgaatt attgcctaaa 2580

gcaatattgg attcttcaca gattcttcta tctatgtgtg gatctcttgt actgattatt 2640

gtagttaacc cttacttctt aattttgata ggagttttaa gtgcagtgtt tggtgtaatg 2700

agacatatat atttaaaatc ctctaagaat attaaaaggt tagaaggaat aatgagaagc 2760

ccagttttta cacatttaag agctacaatt gaaggtctga caactataag agcctttgga 2820

gcgcaaagca ctcttatgga cgaatttgat catcatcaag actatcattc aagtgcatgg 2880

tatatgttta tagtttccag tactgcattt ggattttact tggattgttt ctgtacaata 2940

tttttagcag tacttacttt ttgtttattg ttattcggag aaacttttaa tttaagagga 3000

ggtgaagtag gtttggcgat tactcaagca acagcactgg caaacttttt acaatggggt 3060

atgcgccaat ctgccgaagt aaccaatcaa cttatgagtg tagagagaat attagaatac 3120

aaaagtttag acaaagaacc acaaccagtc ggtcctaaaa aacctgcgaa agaatggcca 3180

cagaaaggag aaattacttt taaagataca tgtttaaggt attttgaagg gggaccattg 3240

gttttgaaac atcttaacct aagcattcaa cccaaggaga aggttggagt tgttggcaga 3300

acaggtgcag gaaaatcttc tttaattcaa gcactattta gactggctcc aattgaagga 3360

actatcaaaa tcgatgagat cgataccaaa gatattaccc tcaatgatct ccgtttgaaa 3420

atctctatta ttccacaaga cccagtccta ttttcaggaa ctttaaggta caacttggac 3480

ccattcgaag aatatacaga cgaagtttta tataaagcta tagaagatgt agagttaaga 3540

gatcctgcta atgttattaa ccgattagaa aatagagtca tggatagagg ttccaattac 3600

agtgttgggc aaagacagtt aatttgctta gcaagagcaa ttctgaaaaa taataaagtt 3660

ctgatgttag atgaagccac agccaatgtg gatccacaaa ctgatgcttt aatccagaaa 3720

actatcagaa agaagttctc agattgcact gtgataaccg tcgcacacag gttaaatacc 3780

atcatggact ctgataaagt attagttatg gatgctgggc aaatatctga gtttgatcac 3840

cctcacttac tgcttcaaaa gaaagatgga gtttttaaaa gcatggtttg tgaaactgga 3900

agagcaacag ctgaacagct caaaaaaatt gcttccgata gttatcaaaa actacaagca 3960

ctaccagagt aa 3972

<210> 23

<211> 4050

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 23

atggattcga ctaagaagca tgtaaaagtg tcaccggaag agagtgctag tccgttttca 60

gtactttttt attggtggat tctgcctttt ttgaagtatg gatatcaaaa tgatattggg 120

atgaaagatg tatataatac cacacaagca gatcaatcag gaccattagg ggatgaatta 180

caagaaaatt gggagcgaga aattttaagt tatattgaca agcagaaaaa caaacctagt 240

ttaaaaaatg caatatttag gacattttgg aaatctttta tcctttccgg tgctgcaata 300

tttgtacaat ttataattat taagacactc cagccagtag ttttggccaa atacattaat 360

ttctttgata caaacaacaa accctacctt ggttggatat ggggatgtgg agtggttttg 420

ttggcattgg ccaatgtggt tctataccat agcacaatgt tagcaaccca aaggattggt 480

atgagaattc gtactgcggt atcttcgttg acttacagaa agcttttaaa attaaaccac 540

aaatctctag gagaaactgc ggcagggcaa ctggttaatc ttatgtccaa tgatgttcaa 600

agattcgatg tgtgtgccgc ttccatacat tttatttgga tcatgcctat atacgcggta 660

cttacattct atatcttgta tatctatgtg ggaatcattg ctgctgtgac tggaatggcg 720

tttattagtt tggaatcaat tccattacaa ggaacaatat ccagatggca gggtgtattg 780

aggtataaaa ttgccctaag aactgataag agaattaaac tgatgagtga actaacatct 840

ggaatacaag taatcaaaat gtacgcatgg gagaaaccct ttgaaaaaat tgtagaaatg 900

tcaagaaaat acgaaataga cgtgatagcc aagacctcat atttgtatgg aattttatca 960

gctactagtg tatttactga acgactgatc ctttacgtaa ctttaatacc atttgtgtta 1020

cttggacata gattgacagg aggcattgca ttttcccttg ctaacctttt taataacata 1080

caacttgtca tggcgattaa ttttccaaga gcactatcat cttataatga agcaaacgtg 1140

tctatagcaa ggttagaaaa attccttcta ttagaagaag ttgaagaaga aacagtagta 1200

aacgaaaagc acggtgatca tgtgggatgt attaatttaa acaatgttac tgccagttgg 1260

tcccctaaat cgatcgtacc aacattgatt gatattgatc tacatttaag atgtggaacc 1320

ttatgttgtg tggtgggtaa tgtaggaagt ggaaaaagta gtctcttgca gttatttcta 1380

agagaattac caataacgtc aggttacatg aatattgcag gaaagatctc atatgcttct 1440

caagagccat ggttgtttgt atcaaacgtg aaagataata ttttatttgg aaaatcattt 1500

aacaaaaaga ggtatcaaga tgttataaaa gtgtgttcgc tagaaagaga tttaaaacag 1560

ttaccgtacg gcgataaaac attggtggga gaaagaggca aatcgttgag tggaggacaa 1620

aaagcaagaa ttaatctagc aagagctgtc tacacagaag ctgatattta tttgtttgac 1680

gacccccttt ctgccgtcga tacaaaagtt ggaaagcatc tatttgatga atgtataacg 1740

aaatatttaa atggaaaaac aagaattttg gtaacacacc aacttcaata catgaagaaa 1800

tcagatctga tcgtcataat aaataatggc aaaatagaca aagtggcaaa atttcatgac 1860

ctaactgagc acgagttgaa tctcctccaa caaaccccag aaattgatga taaagataaa 1920

gagaaaatgc cgactatatt agagacgaaa attccaaaag tttcatcgac tgcaacactt 1980

caatcagcat cttcattagc aagttctata ccaactgaag aaccaggaga aacaggcgaa 2040

ttaatagaaa aaggagattt atcgacttct ctttattggg aatatttccg atctgggact 2100

ggcattggtt tcctgctatt tacaggtttt atgtttattt tttcacaaat cataacaaat 2160

gcttcagatt tgtggttatc tcattggact aatgttgaag ctagaagata tgtctcaact 2220

ttaaacttat ctactgaatt catttctaca acgatttctt ctataaacaa ccataccgtg 2280

acagatatgc tcaagtctgt ggcttttaca aacacaaatc ctcaggaaat tatcacaact 2340

gccgatcctt cagttagtgc ttcaaagttg tcttcaaata tgtcatcact agagttggaa 2400

aattttaaaa gtgaaggata ctatatatgg gtatattcag ctttgatcat agctgtagta 2460

atattacaga tatggagatg ctttttgtac tatcaagtgt gcatgagctc atctaaagca 2520

ttacacaata aaatgtttca taatatactc caagcaccga tgagattttt tgataccaat 2580

ccgtcaggta gaatactaaa tcgtttctcc aaagatatgg gagctgttga cgaattacta 2640

ccaagctgcc aaattgatgc gattcaaatt ttgatggttt ttgttggtat attggtaatg 2700

gttgttatag taaatccttg gatgatcctt acaaccataa ttatcggtcc ggttttgttt 2760

attcttcgaa aaatgtactt aaaaaccgct caatcagtga agagacttga aggaatctca 2820

aaagcgcctg ttttctcgca tatatcagca tcactgtttg gaattaccac tattcgggca 2880

tccaacgctg agaaaatggt cactacagaa tttgatattt tgcaagatca acactccagt 2940

acatggtttt tgtttttggt cagcggaaga gtatttggat tctatctgga tgttatttgc 3000

tgcatatttt tggctattgt tacaattcag tttttacttt ttagagacga gaatacgttg 3060

agtggaaacg taggtttagc catttcccac agctacatct taactggtat ggtacaaatg 3120

ggtatacgcc agagcgtgga agtagccagt catatgataa gcgtcgaacg aattttacag 3180

tataccaaat tagaaaagga tggtgtattt gaatcactgc ctgctaaaaa acctccccga 3240

gactggccta ataaggggaa gattattttt aagaatactt tcttaagata cgcgctcaac 3300

atgacaccct ccctaaaaga tttaagtgta gacattaagt ctggagaaaa ggttggcatt 3360

gtaggtagaa ctggtgccgg aaaatccacc ctaattgctt cattgttccg cttagcaccg 3420

gtcgatggtg aaattattat cgatgatata gaaactggag gaataggact acatgatttg 3480

aggacaaata tttctattat tcctcaagat cccgttttgt tttcagcttc ggtccgttac 3540

aatttagatc ccttcgaaaa gcatagcgat gaaattctat ggaaagcgtt ggaaaatgtg 3600

gaacttaaag gggttgttac agatctaaat caacctgtta gtgaaggagg ttccaatttt 3660

agcgcaggac aaagacaact catatgtctt gcaagagcca tagtacgaaa taataaaatt 3720

cttgtcatgg atgaagctac agcaaacgtg gatccgcaaa ctgatgctct gatccaaaaa 3780

acaattcgtg agagatttag agattgtaca gtactaactg ttgcacatcg attgaacact 3840

attatggatt ctgatcgagt tcttgtcatg gatgccggcc aagctgtaga attcgatcac 3900

gcctaccgac tattacagaa ttcagatggg tattttgcta agtttctgaa agatgctgga 3960

ccaccaatgg caaataagtt aagagatatt gctaaggagg attattataa gaaacacacg 4020

tcgccagaag ttattgtcga tgacgattaa 4050

<210> 24

<211> 1944

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 24

atggttaaag ttccagcaat tggtattgat ttgggcacca cgtattcctg cgtgggggtc 60

tggcagcatg gaaaagtgga aataatcgcc aacgaccaag gcaacagaac aacacccagc 120

tatgttgctt ttacagatac agagcgtctc ttaggagacg ctgcgaagaa tcaagtggcg 180

atgaatccca gcaacacagt ctttgatgcc aaacgtctca tcggcagaaa gtttgacgat 240

ccaaaaatcc agcaagacat ccaacactgg tcatttaaag taattaacca ctgtggcaag 300

ccaaagattc aagttgactg taaaggtgaa caaaaagtat tttctccaga agaaatcagt 360

tccatggttc ttacgaaaat gaaagaaact gcagaagctt atttgggaac cactgttaga 420

gatgcagtta ttacggttcc agcctatttt aacgattccc aaagacaagc cactaaagat 480

gctggtgcca tcgctggtct caacgttttg agaatcatta atgaaccaac agcagcagcc 540

ttagcttatg gtcttgacaa aaatctgaaa ggtgaaaaga atgtactcat cttcgacttg 600

ggtggaggta cctttgatgt ttccattttg acgatcgatg agggctcatt gtttgaagta 660

agatccacag ctggtgacac tcatcttggt ggtgaagact tcgataacag gttagtcaat 720

catttcgctg atgaattcaa gagaaaattc aagaaagatc tgaaaaataa tccaagagcc 780

cttcgacgat taagaacagc tgcagaaaga gccaagcgta ctttgtcatc cagttccgaa 840

gccactatcg aaatagacgc tcttttcgag ggtatcgact tctataccaa aattagcaga 900

gcaaggtttg aagaattgtg ttcagatctc ttcagaggta cactacaacc tgttgagaaa 960

gctctgaatg atgccaaatt agacaaagga caaatccacg acattgttct ggttggtggt 1020

tccacacgca ttcccaaaat ccaacaactt ctgcaaaatt actttagtgg caaaccacta 1080

aatctttcca tcaacccaga cgaagccgta gcatatggag cagctgtgca agccgcagta 1140

ttaagtggag aaaccgattc gaagatccaa gatgttctac tggttgatgt agctccatta 1200

tcattgggta tagaaactgc aggaggagtt atgagcaaca taatttcaag aaattcaaga 1260

attccatgca aacagagtca gacgtttaca acatacgctg acaaccaacc agctgttact 1320

attcaagttt ttgaaggcga gagagccatg acaaaagata acaatcgtct gggaacattt 1380

gatctaactg gcattcctcc tgcgcctcgt ggagttccaa agattgaagt tactttcgac 1440

ctggatgcta atggtatact caacgtttcc gccaaagata caagttctgg aaatacccgc 1500

aacatcacca ttaagaatga caaagggcga ttgtcacaac aagaaattga cagaatgtta 1560

tcagaagccg aaaaatataa ggaagaagac gaacgccaga aagagaaaat agcagcaaga 1620

aaccaacttg aagggtacgt cttccaactg aaacaagctg tacaggactg cggagacaaa 1680

ttatcatctg aagacaagtc agcaatcgag agagaatgcg acagctgtct gaaatggtta 1740

gatagcaaca ctttagctga caaagaagaa tacgaagaca aacagaaaga attgacaaaa 1800

gtatgcagtc ctatcatgag caaattgtac caaggaagtc aaaataacaa tcagcatagt 1860

ggtggaatgc caggtggttg tggacaacaa tacggtggtt ttggtggaag acaaagtggt 1920

cccacagtag aagaagtgga ttaa 1944

<210> 25

<211> 1917

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 25

atggtgctac ccactgactt aactatcatg gacgaccgtt tgagatggaa gcaggttgta 60

cagaaaaaag tacaaaaagc cgtagaagaa gaggtgaaat atttaatatc ggaaaaagga 120

gaaactaaaa tcaaagcaca agtagacaag aaattcttct cccaattatc gcaactattt 180

ggaatcgcct gcccaggatg gaccagccaa gaaagtggct tattcttctt aatcgcctta 240

agtttagtat cccgatctat gtgcgatttg tggctgatca atcatggtac taaaatcgaa 300

agttcaatca tcgcaatgga cccaactctc ttcaagcaac gcctgctcta ctacgtctta 360

gctattccaa taatctcagt tgtaaacaat gtcctgaaat actcaatagg tgctctgaaa 420

atccaactac ggactaacat gacacgacat ctctatgagg aatatctgaa gaattacact 480

tactacagaa tatcaaatct ggacaatagg atatccaacg cagatcaact gttgacgacc 540

gacatagaca agttttgtga aggagttacg gatttgtatt gcaacactgc caaacctttg 600

ctggatattt gtatttatgt ttataagtta tcgacaacat tgggaggggg cactcctgga 660

attatgctgg tgtatctatt agtttctggc gtattattaa cgaatttgag aaaaccaaca 720

gcaagattaa cagcgggaga acaaaaatta gaaggtgaat ttcggcatat caattcacgt 780

ctaatcaccc attcagaaga agtagccttc tacaacggta actctagaga aaaggcaact 840

ctcatggcca gttacaacaa gcttttaaat cacttaagga agttcttaag gtttagagtc 900

gcaatgggcg tcgtggacaa cattgttgct aaatattttg ccggagtagt aggattttgg 960

gtcgtgtcgt tgccatttat gacaagtgga cacagcttta ggcttcttgg aacaaacgaa 1020

agatcaagat tgtattacac gtacggtagg atgttagtaa agttagccga agcaattggt 1080

cgattggtcc tagcaggaag ggatctaacc cgcctagcag gtttcacagc tagagttaca 1140

caattaaaga cagtgttgag tgaacttaac tctggaaaat accaacgaac aatggtttcc 1200

ggatcagaat ctttacagat caacggcgga aaattaattt tcagaaacaa catcattaaa 1260

ttccacaaag ttcctttaat aacccccaat ggagacgttc tcattaacga aatcactttt 1320

gaaattaact ccggcatgaa tgtgttggta tgtggtccca atggggccgg aaagtcttcc 1380

ttgtttagaa ttttaggaga gctttggcct ctgttcggtg gtgagctcac gaaaccacct 1440

agggggaaat tattctatat cccacagaga ccgtacatga cgttaggatg cctcagggat 1500

caactgacat acccgcattc tggggcagaa gcagcgcgaa gaggaacgac agatgccaag 1560

ttggaggagt atttgcaaag ggtgcaactc ggatacattt tggaaagaga gggtggtcta 1620

gatgctgtag cggactggtt agatgttttg agtggtggag agaagcaaag gatagctatg 1680

gccagattat tctaccacca acctcaattc gccattttgg acgaatgcac aagcgctgta 1740

tctgtcgatg tagagggcag tatgtataag tattgtagag atgttggcat cacgttactt 1800

acagtatcac atagaaaatc actttggcag catcacgaat acgtattaca tttagacggg 1860

cgaggaggat atagttttaa accaattgat aattgtgatg aacaattcgg atcatag 1917

<210> 26

<211> 2265

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 26

atgccaacag taatctccaa gttcctggaa cagggtgaaa gcaccttcaa gcagaacaaa 60

tccgtcttct ccggagcatt catcgcagcc gttctttgca cctacgccta caaagtcggc 120

tatcccttcg tggactcctt gatacacaaa ccaaaggaca atctcaacaa caaccacata 180

gttcaaaaag acttgcaacg taagaatggc ctagtgaaga acaagaaact caaaggaagg 240

cttaaaaata gtataccgaa cttcaattta gcctttatct tacaatttat taagttagtt 300

aggataatga tacccagttt catttgtact gaaacggttt tactcagcgg gcatacgacg 360

tttttgtttt tgaggacgtt tttgagcatt tatgtcgcta atttagaagg tgctatagtt 420

aagtatattg tgatgaaaga cccccagaac ttcgttaagc agctaggaaa atggtttgcc 480

gtagcgattc cagctacgtt tattaacagc atgataagat atcttgaaag tcggatagcc 540

ttgagtttcc gcacaagact ggtagaccac tcgtacaaac tatacttcaa gaaccagagt 600

tactacaggg tgactgtgct agacggcaga ttagacaatt gcgcgcagcg gctgacagat 660

gacatagaaa ctgtcgccaa caccgtgtcg cacctttacg gacagataac gaagccatgt 720

ttcgatatac ttctcatggt tatagcttta gccaatttgg tgaagagtcg ccattcgaat 780

cttgtgatag gacctgtgat tatttgcgga gtggtgatgt tctcagcttt acttttaagg 840

tttgtgtctc caaggttcgg ccacttggta gcccaagaag cggagaagaa aggatatcta 900

cgacacgtcc acggaagaat cgtcagcaat gccgaagaaa tcgctttcta cggaggccat 960

caggtcgaag aaagtcaact tagacaagct tttagagttt tatctaaaca cttggaacac 1020

atgttcggtg tgaaattatg gttcatcatg ttggagcaat tcttgatgaa atatgtgtgg 1080

tctggagcag gtataatagt cgtatctctg cctattctcc tagcagccgg caatcgacgc 1140

aaaatctcca acaaaagtct tctttccata ccagattttt catccaaatc agaatctctc 1200

atcgctgaag acacagccga cgaccaaata gaagatagcg tatcagaacg aacgcactac 1260

ttcactacct ccaagaacct gttgataact ggctcagatg ctgtagaacg gctcatgagc 1320

agctacaaga acatcgtcga gttagctggt catacggcca gggtggccaa catgttcgag 1380

gttttagagg aagctagtaa cggaatttac cacaagactc tcgtcgctaa gaaagagaag 1440

tctgctgatt tcgagattga gttccgtggt gatcaacctt tagccaaagg taaaatcatc 1500

tactcaacca ataacgagat catcctaaag aatgttccta tagtaactcc aaattgcgat 1560

atagtgtgtc cgtctctaag tcttgaattg actccgggac aacacttgct aattacgggt 1620

ccaaatggct gtggaaagtc cagtttgttc agaatattga gtggattatg gcctatttac 1680

ggtggagagc ttcatactcc taaaaattca atgttttata ttcctcagag gccttatatg 1740

gtaataggta atttaagaga ccaagtaata taccccgaca cttacacgga tatggtaaag 1800

aaaggtatta ccgaagaaaa tttactgaaa ataatgacta tggtacatct agatcatatc 1860

gtcgagaggg acggattcta tgaaatgaaa gattggactg acatcttatc cggtggtgaa 1920

aaacaaagga tggctattgc gcgattgttt tatcacaaac cgaaatacgc tctcttggac 1980

gaatgcactt ctgcagtctc cattgatgtt gaaagcttca tataccaaag tgctatagac 2040

atgggaatta ctctactcac catcacacat agacctacac tatggaaatt ccatacgcac 2100

atccttcaat tcgacggtac aggatcatgg gaatttagtc agttgaacca caccagcaga 2160

cttaccttga agaaagaaaa ggaggacctt ctaaaagcag aaaacaacga agaacgatcg 2220

aaaagactgg acgaactaaa caaattactt ggagaggatt cataa 2265

<210> 27

<211> 2766

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 27

atgtcgaaaa aacgtggggc caagaaggga aaaaatttgg atgatgattt tgaagaaacc 60

tccagtatta ttagcgaaaa agaaaaaata aattcgaaaa ctcccaaaaa taagactact 120

aaaaaaggga aaaaaggaaa ggatgactgg agtgatgacg aggaaattga acaaggcaaa 180

gaagttaatc aatctgaacc agcggaaagt aaaccagtag ccaagaaaaa gggcaaaaag 240

ggcaaaggca ataagaatga tgactggtca gacaaggaag aagcggatat aaaactttca 300

gacagtgaaa ctgagtcaat gccagtagca gtgaaaaaat ccgctaagaa aaataagaaa 360

aagaaagatg attggtcaga taaagaggtt gacatacaat tatctgaaag tgatattgaa 420

gaggcaccta aaattgttaa aaagtcagct aagaaaaata agaaaaaaga tgattggtct 480

gataaagaag atatagaaat agaaactaaa ttatctgata gtgaagaaga actagcacca 540

gctattgtta aaaagtctgc taagaaaaat aaaaagaata aaaatcaggt acgtgatgaa 600

tctccagata ttgaggaaat ggaagcacac gatgaaggtt ttaaggagga tgacgtaagc 660

gaggaagaaa ttgtaaaacc tgttaaacaa gacaaaaaga aaagcaaagg caaagaatca 720

aagaaggatg aaagtaaagt tgatgtagaa gttaaagaag aaattaagga atctaaagaa 780

gattccccta tagtagaaga caaagtaata gaaaatggca aacctgaaaa acaaaacaag 840

gagctagaaa ataaaaagga taatgatgta gaagaattaa cagaaaaaat aagtactacg 900

caaataagtg atgataaatt agatgaatct aaagaaaaga aactcacgca taaagaaaag 960

aaaaaaatga aaaaactaca ggaatacgaa aaacaaatgg aaaccatgtt gaaaaagggt 1020

ggacaggggc attctgaatt agatagtaat tttactgtat cacaaactca aaagactgct 1080

ggacaattgg ctgcatttga gaatgcagta gatataaaag ttgaaaattt tagtatttct 1140

gctaaaggta atgatttatt tgtgaatgcc aatttattga tagcccaggg cagacattat 1200

ggtttagttg gacctaatgg ccatggtaaa acaacactgc tgagacacgt agcacaacgt 1260

gcatttgata ttcccccaaa tatcgatatt ctctattgtg aacaagaagt cgtggcagat 1320

gataatactg ctgttgaaac cgtcctagca gccgatgtca aacgtaatga tttattagcc 1380

gaatgtaaaa aattagaagc cgctgctaat agtggagatt tagaaattca ggagaggctt 1440

aacgaagttt acagtgaact taaagctata ggggccgatt ctgctgaacc tagggctagg 1500

agaattttag ctggtttagg attcgataag gaaatgcagg atcgtgctac caaaaatttc 1560

tctggaggtt ggagaatgag agtatcttta gcaagagcat tgtatattga accaacgtta 1620

cttttgcttg atgaaccgac taaccattta gatttaaatg ctgtaatttg gcttgacaat 1680

tacttgcaag cttggaaaaa gactctgttg atcgtttccc acgaccagtc tttcttagac 1740

aacgtctgca atgaaattat tcatttggat aacaagaaat tatactatta taaaggaaat 1800

tattccatgt ttaagaagat gcatgtgcaa aagaagaggg agatgatcaa agagtatgag 1860

aagcaagaaa agaggataaa ggaattgaag tcgtcaggtt cttccaaaaa acaggcggaa 1920

aagaaacaaa aagaagccct tactagaaag caggagaaga atagaactaa gattcagaaa 1980

caagaagagg atacaactcc cacagaacta ttacaaagac ctaaagacta tttggtcaaa 2040

tttcgcttcc cagaacctcc acctttgcag ccacctgttt taggattaca caatacaaga 2100

tttgcctatc ctggacaaaa acctctattt gtagatacag attttggtat tgatatgagc 2160

agtagagtcg caatcgtagg tccaaacggt gttggcaaat caacattctt gaagttatta 2220

acaggagatc tatctcctga taaaggagaa aataggaaaa accatagatt gcgaattggt 2280

agattcgatc agcactctgg tgaacactta acggctgaag aaacacctag tgaatatctg 2340

atgcgtcttt tcgaccttcc ctatgaaaag gctcgtaaac aattaggaac attcggatta 2400

gctagtcacg cacataccat taagatgaaa gatttatccg gtggtcaaaa agcaagagtg 2460

gccttggcag aactctgttt gaatgctccc gatgtattaa ttttggatga accgacgaac 2520

aatttagata tagaatccat agatgcctta gctgaagcaa ttaacgaata taccggagga 2580

gtcataattg tgtctcacga cgagcgactt atcagggaaa ccaattgctc tttatatgtc 2640

atagaagatc aaaccatcaa cgaattagaa ggagacttcg atgattacag gaaggaatta 2700

ttggaaagtt tgggagaagt catcaacagt cccagtatag cagccaatgc agcggttgcc 2760

caataa 2766

<210> 28

<211> 1993

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 28

atacggtctt ttgggagcta gtggttgcgg aaagactaca ttattaagca gcatagtagg 60

ccgaaagaaa atagatggag gtgaaatttg ggttttgggt ggaaaaccgg gagaagctgg 120

cagcggagta cctggaccca gagttggata tatgccacag gacattgctc ttgttggcga 180

gtttacagta aaagatgcga tctactattt tggaagaatc ttttcgatgg aagatagttt 240

aatagcaaaa cgttatcgaa atcttcatac tcttttagag ttgcctccag atgataggta 300

tttaaaaaat tgtagtggag gtcaacaaag gagagtttcc ctagcagctt ctttggtcca 360

taaacctgaa cttttaataa tggacgaacc cacggttgga gtggatccag tgttaagaga 420

tcgtatatgg aagcatctag tagatattac taaaaaggat aatacgtctg tgataattac 480

tacgcactac atagaggaat gtagacaggc aaataagatt ggtctaatgc gagagggaaa 540

acttttagca gaggagtcac caactaggtt attaacattg tttaatagtg agactttgga 600

agacgtattt ttgttgctca gcaaaagaca ggaagaagga cgattacaag aactaacctc 660

acatcgagtt gtagatgatc agaataattc tatgttagct aatgacaccg caggatccac 720

cactagcgta gcaacgtcgg tttctacatt tgaaatcggt catggttcca ctgatatttt 780

agccaaaaag aaaatactga aggcaagaaa tgccttaaat aaaagtcgaa tgaaagcatt 840

gtttgataaa aatttaaaac aattttatag gaacatcacg ggtattatat ttttaatgac 900

atttccaatt ctacaagtag gggtatttat gggagccgta ggtggagaca ttagaagcat 960

acccttggga atcgttaacg acgaagcgat gagcgttaca tgcccaggct tttcatttaa 1020

cggtacggca actgctacag acgatagagc ctgtcagctt aggaatatca gctgtcggtt 1080

tttatcgtat ctggaccatc ccatgataga aaaggttcat tttgagactc tcgaggatgc 1140

caaagatgca gtcttgcatg gcaagatagt aggagctatg tacatgtctt cgaattttac 1200

atcattttta gaagaaagaa tcgataaagg gaaggatatc gagaaagata ttttgtcttt 1260

tagtgagatt aaagtttgga tggatatgtc aagtagatcg ccaatcggag ctaccttaaa 1320

atacaagctc attgatttat acaccaaatt ccaaaacagc ttatttgatg attgtgattt 1380

tgttcctggc tttggcgact tgcctgttaa cataaatttc atttatggag atggagatga 1440

gccgtacaca gtttttatga taccgggaag tcttattacc atcatgttct ttatgggtgc 1500

aattatgaca tcccagataa tcattaccga ccgacacgac ggtgtgtggg acagatccat 1560

tgttgcagga gtaacatctt tagagattac cattacgcat ttggtgttgc aggcttcgat 1620

ttgcataatt caaacggcag agctattggt tgtcgtctat ttgatttacc aacaagaata 1680

ttcaggaagt ttgtggttaa tgtacgtgat ggtctatctt caaggtattt gtggaatggc 1740

gtatggtttt tgggtatctg ttatcagcac tgatcattcc atggccaata ctgttctgac 1800

aggaatattt cttccaatga tgatgttaag tgcagatacc gctacgcacc tactaccttc 1860

acgtcagttg caatggggga ctaatagatg tcgaaagttt ttacctggag tagagaaagc 1920

agcctatgca ttctctgaac agcctctaac aagaggtgaa atcgtcgata agagtgctgg 1980

ctgcactctc taa 1993

<210> 29

<211> 2484

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 29

atggtcctac ctacaattgc aaatctcgag caaaacccag aggctctccc gttcaccgaa 60

gacttcaaca aactcttcga agctttcttc cagacttatt ccagacagaa ggaacttgaa 120

acgcttgccg aagacatgac atcggatcta acggacaaag cccacaaaat cgagctgtcc 180

ttgaaactag ccgatgaaga tcagaggacc atcatcgagc tacgacggca gatagaaaac 240

gcttggaagc tagctgatgc tgctcatgct agagagcagc tggctcatga gattattgac 300

aatttgaggc gacaggtgga gaatttgaat gctgagattg agtttaagaa caaaatgaat 360

caggatacgg atgaatacgg acttttgggt gctagtggct gtggtaaaac gactttgctc 420

aattgtattg taggtagaaa aagacttaat tccggagaac tatgggtact gggtggtacg 480

cctggttcga gaggcagtgg agttcctggt ccacgggtag gatatatgcc acaggagaca 540

gcactgaacg gcgaattcac aatccgagaa actctaaaat acttcggctg gatatcccgc 600

atgaccacag acgaagtaga agccaaagtt gattttttca tcaacttctt gatgttaccg 660

gacgcagata ggcaggtcaa aaccctcagc ggagggcaac agaggcgagt ttccttagca 720

gctacgcttc ttcacgaacc tgagttacta attttggacg agcccaccgt tggtgttgat 780

cctttgcttc gaacgaatat ttggaaccac ttggttgaaa taactaaata tggtaggacg 840

actgtgatca ttaccacgca ttatattgat gagactaggc aggctcattt gattggtctt 900

atgcgtggag gctacttctt ggccgaggaa tccccagaac gactgataac acaattcggg 960

gtggatagtt tggaagacgt cttcttgaaa ctcagcgtga tgcaaaacat gggaaaacga 1020

agacgctcca gtatagcaaa aagtgtcgtc gagacgataa cggtgcctga actaggtgga 1080

gctgttaatc ctgctgctgt gttggacgac gacttgggag agatttctgg ggaattcgga 1140

gacagtgttt cggttactag taggacagga agaagagtgt ctattgctcc tgaacccacg 1200

gcagaacaca ttattccaga actaccacca gacgaggaac cggaagtgtc cttcaaagat 1260

tactttaaat ttgtccaatc gcatcacatg cgcgccttga tatggaaaaa tttcctgtgg 1320

atgtggagaa atattcccat gatgatgttc attatcggtt taccaataag tcagaccatt 1380

ttgttctgtt ggactattgg acacgaccct caaggaataa aagtatcagt tgtcaataac 1440

gagattaatt tccctgatga gacttgtcat cagcaacctc taaattgtaa tggtacaaaa 1500

ataagttgta attacttgaa cgaagtggct aagagttatt ctataacgtg ggaatttatg 1560

aaaacggaag aagaagcgag acatagagtg gaacgaggaa agtcctgggg tgttgtagtc 1620

gttccacaca atttcaccga cgcgctttgg agtagaattg aaaattcgag agatactcct 1680

ccagaagata taatggcttc tactatcagt gtctatgaag ataaatcaaa tgaaaacatc 1740

gcaactttcc ttacacgaga tatgctgtat ggattcctca cctttatatc tgaatttgtg 1800

aaaagttgtg gatataatga aaaaagtgtt ggcgttccta ttcgatacaa tgaaccaatt 1860

tatggatacc aagacccaga ttttaccgat tttgcagcac ctggagtagt cttgactatc 1920

atcttcttca tggctgttgc cctcacgtcc ggttcaatga tcatggaaag gaacgaagga 1980

atcttggaaa gatctttggt caacggcatc agcggaacgg agctgttgtt ttcgcaaatt 2040

atcacccagt tcgtcgtcat gttgggacag tccatcgcag tacttcttgt cgctttcatt 2100

ctctttggtt taacgcaacg aggagattgg acgacagtga caaccttgac tgtactggcc 2160

ggtatatgtg gaatgtgttt tggtttcgtg gtggcctgca gttgtgaaaa tgaaagatct 2220

gctacctacc tagctatggg cagtttctta ccgatcgtaa tgttgtgtgg catcatttgg 2280

cccatagaag ccatgcataa gtacatatcg tggatgtcgg tggtactacc tttgacccaa 2340

gccacagaga gtttgagatg tatactagca agaggatggc ccatcagtaa tcccacagtt 2400

tacaacggtt ttatatcgtt gaccatatgg atctttatat ttttgacgat cagtattctg 2460

ttacttaaat tcaagaaggg ttag 2484

<210> 30

<211> 1842

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 30

atgggtgaaa atacagtttg tgatatctta ggaactgtgg atccacctca agagagcaat 60

aagatacatg acgaagactt ccgtttgcat tctttggaac taccagaaaa tcaagttaat 120

gatccagcgt ctctaataaa tgtaggattt agacatatat cttacagtgt ttatgacgga 180

atattatttt caagaacgaa aaggaaactg ctggatgata tcacaggaaa ttttccagga 240

gggcaattga cggctataat gggaccttca ggtgctggaa agacatgtct aatgaatatt 300

ttatcaggct atgcgactac aggcatactg ggtgaagttt tagtcaatga tgaaccaaga 360

aacaatgttg cttttaggaa acaatcgtgt tatataatgc aaaatgatga tctccaacct 420

ttactaacgg tattagaatc catgagagtg gctgctgatt taatgcttac ggcgtctaag 480

actgacaaag ataaaaagat agcatccatt ttgaaatccg tcactctttg ggaaataaaa 540

cacaccagga cagacgccct atctggggga cagaagaaac gtctttctgt agcattagag 600

ctgcttagag atccacaagt tatgtttttt gatgaaccta ctagtggatt agatagttta 660

aattcaattc gtcttgtaaa attgctgaaa gaaatgtccg aaagcggtaa aaccattatt 720

tgtacgattc atcaaccagt gccacgatat ttaaattgtt cgatcacctt attggaagta 780

gcttcaggag attatgggga ttttacacat gctctctcaa ctaagtcgaa taatggaatc 840

aatgaaagtt atgcgaaatt aagtgctagt acaatgaaac taaattcaat aaaagaaccc 900

aatattggag aagattttct tgcagcaaca aaaagaacaa tcgtaaacga aatcgaatat 960

acttgggatg agtgcggctt aaattcttat ccaacttcaa cgataaacca gtttgtggtg 1020

ctgacgaaga gatcttttct aatgctctct agagacagaa ctttgacgta cttcagattg 1080

gggactcatt ctgccatagc gctattttta ggaattttgt acttcggtgt tgggctggat 1140

gccgctaata ttaatgacaa ttttagtttt atgtttttta cggttatgtt ccttatgatg 1200

acggcgttca actgtgtagt taccacattt ccttcagaat tacctataat aattaaggaa 1260

cactttaata aatggtatgc catacgatcg tattatgcag cagtatctat ttcagatata 1320

accgtccaaa ttttggctac cgttttgtat gccacaataa cctatttatg tacacaacaa 1380

cctatggaat ggagtagagt atcttcgttt ttgtttatat gcattttggt ctcggttatc 1440

tcccaaagtt gggcattggt tgtgggctcc tgtttgagtg taacgaacgg cgtagtaata 1500

ggtcccttct ttcttctacc attcatcatg ttttccggat acttcgttca acttcgtgat 1560

tgtccgcacc aacttaaatg gatgtttgac atttctttcc cccgatatgc gcttgaaggg 1620

ttggtactca cgatattcgg ctatgacaga ggaaaactac cctgcgagtc caaggatttc 1680

tgtctctacg tatatcctga gcagtttatt aaagacaaag acatggaaaa tgccagctat 1740

acgataggag taatgtgttt gatgggttta attgttggga tgaggattgc aggaggagct 1800

gctttgagta taaggctgag gcatgacaga cgaaggagat ag 1842

<210> 31

<211> 1878

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 31

gtagaaaagc gctctgttct tcaaagaatg gttggcaaca ttaaaactgc agtaatctta 60

aaagatgtgt ccatgttggt acattctggt gaggtcctag cagtgctggg atcaaaaggt 120

agtgggaaga aagccttact tgatgttatt tcaagacgag cacaaggccc tataagagga 180

cagatatatc tgaacaacca tccagtcagt atgtgtttat tccagcagaa atgtgcatat 240

gttacacata aatgtgattt tataccagga ctaaatgttg aacagacgtt gtattatact 300

ccgactaagt tcactggata tttaaaaatg tctaaagtaa agcaagtgat agcagattta 360

gcattgtcac aagtcgctaa aaaatgcgtt gaagatttga caaaaagtga atacagaaga 420

ctgatgattg gtgtccaact aatcaaaaac ccagtggtac tattactaga cgaacccacc 480

tgggatctgg acccactaaa cacataccta ataatttcaa tactttcaaa tgcagctaaa 540

aagtatggaa ctgctataat cttgacgatg gagaagccta gatctgatgt tttcccgttc 600

ttggatcggg tgttgtactt gtgtcttgga gatgtcgtct acactggagg taccaagcaa 660

atgctcgagt acttcaacgt tattggcttt ccttgtccac aactggaaaa tcctttgatg 720

tactatcttt gtctctctac agtggacaga agatccagag aacgattcgt agaaagcaat 780

taccaaatag cagcattagt ggagaagttt aaaaacgaag gagttatttt ccagaaaagt 840

ccaagcatga tctcgcccaa tccgaaccac gagcatggac aacacgagaa ggttccattc 900

atgcatggaa gacctggaag gttttcaact gggtggacga tttacgtgcg acttttagcg 960

gcaactgtga gttttaaaag agctggactg cgacaaacgt ttttgcgtac ttttgcgcta 1020

cctttatatt tcttcttgat gtggctgttc tatagagaaa tgaaggattg gcaacataca 1080

ttcatatcaa gaaatggctt aattttgaac tgtctgtgct gtgtttactt cgttggaata 1140

ataaacacca tattaatcta tcctatatat aggacgaggt actaccaaga tactcaagaa 1200

ggattatatg gaggcacgtt gtttctgcta acctacaatt tggtttcgct gcctttctca 1260

tttctttcga cagttttgtc tgcagttata attttccgta agttaattat atcattcgac 1320

aatcctgtcg atttcgtgta cttcactcta atattatggg cttgttacat cttcgcagaa 1380

cagcaaacca tggcgatttt aatgattgtg aaggattaca tgaaagccgc tatatttagc 1440

atttatctga cagtggtgtg cattactctt ggcagcggga tattaaggtc aatgaaaggc 1500

ttaccagaat ggctgtatta cgccacctac gggactcaag cacgctacgc tgctgcctat 1560

ctaaatcgca aagtgtttac ccactctgcc ctcagcaaag cccttccttt tgatctcctg 1620

cacaactgca ctcaaatcag cttcgagact tccatcttaa acggcgccaa caatgcttac 1680

tgtagatacg ccagtggaca agccttcctc acggaacgat acagcagaga tcccacggag 1740

gtcattttta gcggagttct ggaagaagat ttcaatgttg gactcacatg tgggtttgca 1800

ctgggaatga tcgtttttaa tttgtttttg tatctgattc ctttgccttc gttcgttaag 1860

gcaaagttta gggaatag 1878

<210> 32

<211> 2823

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 32

atgatgcgta gtcgcacccc ctcccaagga gggggctcgc aaggattcga aatggagagg 60

aaatactcgg taccttctaa cccggaatca agagcctttt cgggtggaac taccagtgaa 120

gatctgcacg cttggtcgat atataggcaa aatctaaatt cagatttcac tgatagtgct 180

ttgggatcca cagataagag tccacttcct tatggcaatt ttcagcttcg agatacaacg 240

gtgcagtcta tcctctctca tcctcgttat ggtcctaagt cagccctggg ctccaacatg 300

tacacctacc taaaattcgg ccttccgagg gtgttcccac cgaaccataa tggttctaat 360

cggtctggta ctccacaaca ttttcgacga aactcgtcca ctagaccgca taacattcgg 420

cgttcgaaag caggaagtca tggaccaaga gatggaagtt ctggatatga tagttctgac 480

aatgaaacga gccacaacta taagcaaaat cgaaaatatc gatcggaccc tgatttcaga 540

atgcaaaatg tgtatcacgc tgagcaaacg agtcctggga tcccattggc agccatgcac 600

caaggtgata tacgacactc aaacagtcaa tggaatagaa ataagagcat ttcggaagcc 660

aatcttttgg ctttaggtta taatagatcc tgtcataata gcgaacgcca tctaatagat 720

cctcgaagaa acagtgttgc cgattatgga caccacaatc acgatgtcgt ggatcatagt 780

gtcatgggtc atatgggaag accaatgtcc aaaactgaaa gtcatttctc tgtgcctcat 840

tcaagaagag gaccaccatc tattcttcga accgattatc tcaatcaaga tgatgaatct 900

ggaaccacgt ttatgtttcc acatttacag gcacatggat taggcatttt cccaagctcc 960

cagtcttgca ccaaatcccg tcaacacctt cttcttaacg aaatatcgtt tgaaattcga 1020

gggggcgaaa taatggccat tatgacgact tccgaggaag aaggtaccgc cttgttggac 1080

atcattgctg ggttctcgtc accagcttta ggaacaatat ttttaaatgg acatagcgtc 1140

agagcgcata cattgaaatc tagagtagct tatgtccaaa atgacttgaa tttatgtaag 1200

gatatgacag ttgtacagac tttaagatta cattatgatc ttaagaaacc aacagagaaa 1260

ttagggtatt taaaaattga atctatggat aggataaacg ttttaataga cgatctaggc 1320

ttagagcaag tccgaaacac caaggtatct atgatgacaa tctcagagag aagaagactg 1380

aacgtagcgt gtcaccttat cctcgacact gatatcgtac tcttagacca acccaccaaa 1440

agcatggaca ttttcgatac ctttttttta gtcgaatatt taaaacaatg ggcgagcggt 1500

ggggcaggta gtactttggg gcggatagtg atattgacga tgcatccccc cacttacgag 1560

atatttacta tgctgtctag gattttgttg gtgtcggcgg ggaggactat gtatagtgga 1620

agacgacggg acatgctgcc atattttgct ctagttgagt atccctgtcc ggcttttaag 1680

aatccctcgg attactacct tgatctcgta accttggacg atctcgcagc ggaggctatg 1740

ttagaatcat cacaacgaat tgaacagttg gctgaaatat ttagacaaaa acaggaaccc 1800

ttaagcgacc caggaccacc ttcatctttg ccgctcactg tacgaaactg taattgtttt 1860

gtagctgcat ttgctttgtt tacgaaatcc atgatctaca cccaacccgc gactttcctc 1920

agctggctca cagtgatagt cctttcagcc agtctatccc ttattcttgg cgccatattt 1980

tgggatatcc cttccaccga tcctcaactt atattaaacg acaggtacgg gtaccactac 2040

agcgtcatgt gcattgtaca ttggccatta ttgctagcga tgactgtcaa tgaagttaga 2100

agaaatagaa aggtgattga gagagatatt aaagatggac tgtatggtag agtcacctat 2160

atcataacaa aatccatcat caatatattt ccatcattgt ttgtatggct gatctatgta 2220

gtccctagtt actctatgac tggcttatac atgcagcatt tgaataatta cgatggtttc 2280

tatatttata taggggtaat gttgttatat cttagctgca tccaaatatt tttgatggca 2340

tttatctaca cagtcccttt gagcaacaca gccacaatat tctgcggtac agtactctca 2400

gctttcttcc tgagtgctgg atattcattg cacctcaaag atattccaat gtatttacaa 2460

tggattgaaa aaataagccc aagcgagtgg ttgataccat atttactaaa tagagaactt 2520

tctacagaag ccatacagag tttgcaaggg acaataacca ccctttgcag aaataaacag 2580

attcaacacc aggatattat agtccaattg ccatgtccac ctccaaatgg aacaaatagt 2640

ctaaaaagct ttggttactt aaagtctgat aacttaacct tcgactatgg taacccagtt 2700

atagcaatgg gcgtatttta ctgtatcttc ttcgtgatct cttgcttcat gttcgctctc 2760

aatttgtgca gaagcagacg aaggcgtaga caagacacta agaacgatgc aaataaacca 2820

tga 2823

<210> 33

<211> 2037

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 33

atgttggtgc ttagtgttag tggctcccaa accaacatat acaatggttc ctcaccaata 60

accatcagcg gagcgggtgc gctccgtaag gtccccaaca gcagtccgag caaccataaa 120

aggccaatga ttgccctcac acacctaccc aaaaggccgc ccgtggacat taccttcaca 180

gatctctcct actcggtatc cgaaggaagg aaaagagggt ataaaacgat attgaaatgc 240

atcagtggaa aatgcaagtc gggagagttg acggccatta tgggaccatc aggggccgga 300

aagagtacca ttatgaatat tttagctggc tacaaaacgt caaatctgag tgggcaggtt 360

atgatcaacg gcaaagagcg caacctgcga cgcttcagga agatgtcctg ttacatcatg 420

caggacgact gcctctcccc acatctcacc gtcaaggagg ccatgatggt atcagcgaat 480

ctcaaattag gaaagactgt cacactgtcc gagaagaaag ttgtgatcaa cgaaattatt 540

gaaaatttag gacttcaaac ttgcatcgat ataaactcct ccaatctctc ggggggtcaa 600

cggaaacgtt tgtcaatagg tttggaatta gtaaataacc caccagtaat gtttttcgac 660

gaaccaacga gtggtctgga cagttcatca tgtttccagt gcctctgcct actcaagtca 720

ctcgcacgag gaggtcgaac aataatatgc actatacatc agccatcagc taggttattt 780

gaaatgttcg accatcttta catgatggcg gaaggacagt gtatttatag agggcctgtg 840

ctaggattgg tgccattttt gtccagcatg ggcttgaatt gtcccagtta tcataatcct 900

gctgattacg ttatggaggt agcatgtgga gagcatggag actacgttca gaaactggtg 960

gtggcagtaa acgctggtcg ttgcaccaaa ttcgccacgc cagatcatag aagtagtaag 1020

atagtttcca atgatattgc taaagaagcc aacgggaaga actcctcggg agatgtaata 1080

tccgtgccaa atggttctgt gaagcctacc acccccacaa cgccagtcac ttgcacaacg 1140

tcacttttag attcttcgga gaatttgtcg ccaacggaaa agaatggttt ttcaacgaca 1200

ggtctccaac agttcacaat attgctcaaa agaagtatgt acatgattct aatggacaaa 1260

acgctaacta ggatgagact ggtgtcccac tttgtgatag gatgtcttat aggcctaatt 1320

tactatgata taggacaaga tgctgccaaa gttaccagta acgcagggtg cctcttcttc 1380

tgcgtcatgt ttatgatgta cacagctatg atgcccacaa ttttaacatt tcctttggaa 1440

atgtcagtta ccgtaaggga gcacctgaac tattggtact cgctcaaagc gtactatatg 1500

gctaaaacat tggcggacat accgtttcag gtggttatga cgttgtgcta tataataggt 1560

gtttacttta taacatccca accgttagat gtcactagat ttgggatgat attgttagtt 1620

acagtgttaa ctgcccttgt atctcaaagc ttcggattgc tcataggtgc tgcgtttaat 1680

attgagggag gtgtattcct cggaccaatc tccaccattc ccatggtgct cttctcaggc 1740

ttcttcacaa acctgaacga cattcccttc tacctgagat ggctgccata cctgagttac 1800

ctgaaatacg gcttcgaagc gtgtatgatc gccatctacg gcttggatag accgaagctg 1860

acgtgcaaca tcgagtactg ccactttaag taccccaaga agttcttgga gcagatgtcc 1920

atgaaagacg acatggtcag ctatttcatc gacgtcgggg tgctgggagg actgttcatc 1980

tttcttaggg tgatagcgta ctttatgctt cgcattaaat tgatgcaaaa tcggtaa 2037

<210> 34

<211> 1650

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 34

atgggtccct caggagctgg caaaacctcg ttgcttaata ttctcacagg ataccaggtc 60

actggaacta ctggtaccat aaaatgcaat agctctagta gaaagcaaaa aggcgtctta 120

caatacaaaa aagaatcatg ttacatcctc caagatgata gcctaccaaa tttgtttaca 180

gttgaggagt gtatgatgat tgccagtaaa cttaaaattg ccaatatggc caaaaaagca 240

agggaatttt tgattaacga aatcctaacc aatctaagct tattaaaggc aaagaatact 300

agatgccaat ctctatctgg tggtcaaaag aaaaggttat caatagcttt ggaattagtt 360

gataatccac cgattctatt tttggacgaa ccaactacag gtcttgacag cgcatccacc 420

acccaatgtg tagatttgtt aaaaaaatta gccaatggag gaagaattgt aatctgcact 480

attcatcaac ccaacaccca aacttatgaa atgttcgatc aggtttatat gttggccaaa 540

ggtcgatgtg tttaccaagg gccaagtacc aatactgttc cttttttagc ttcagtaggg 600

ttgcactgtc ctcaatacca caaccctgcg gattatatta tggaagtcgt tagtggagaa 660

tatggtgatc atatagatca attagcagta gcagcacagg ataaaaagtg gcaaaatata 720

cctacaatta aacttagtga tactccagct atagacagta aagataataa tatcatatac 780

tccgacgaaa acgttactct ctctaaatca ccttcagaat ggaaaaggtt ttttatatta 840

ttacaacgaa gttctgttca actttataga gattggacga tatctcaact aaaactggta 900

cttcatttgt tagtaggatt atttttaggt ataactttcc aaaattgtgg tagggatgcc 960

acaaaagtga taagtaattt aggattttta caagtcggta tagtttattt ggcttatacg 1020

tcaatgatgc ctgcagtatt aaaatttcct accgaattag taatattgaa gaaagaaagt 1080

ttcaataatt ggtacaaact gacaacgtat tacgctgcct ttttggtttt cgacatacct 1140

caacagatgt tatttagcac agtttactgt attggatgct attttgtcag tgaccaacca 1200

ttggaggttg accggttttt cagtgtactt tttgttctag tgctagcatc attgtcctcg 1260

tcaggatttg gactgatatt gggaacaatc actaatccaa ttaacggagt attttttgga 1320

gcagtgggat tatgcttctt tatctcagtt ggaggatttt tcataatgtt cacgcatatg 1380

tcgaatgtaa tgtacttatt ttcgtatatt tcgtatatca gtttctcagt agaaggagtg 1440

atgcaagcta tatatggata cggcagaggc cagcttcatt gtccagaaga agcagagttc 1500

tgtcagtacg tttcatcaga ggtgcttctc gaagatatcg gaatgagtaa acctaactat 1560

tggatagaca ttatctacct tacttgcaca ttcctaactt tcagaaccat cgcctttgta 1620

acacttaaaa gaaaattagc taatccgtaa 1650

<210> 35

<211> 1815

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 35

tccaaattaa ttttaagaag cgttaatgga agtttccagg ctggacaatt gacagcaatc 60

atgggacctt ctggagctgg aaagagcaca ttgttaaata tactagcagg atataaaaca 120

cagggagcga cagggcaaat cttgataaat ggagcgacta gaaatttaaa acaatttagg 180

aaaatgtcta ggtatattat gcaagaggat atgatccaac cattgttgac tgtggaagaa 240

gcgatgatga tagcagcaaa tttaaagtta ggaaacacgt tgagtctaag cgataaatct 300

tcagcgatag atgaaatttt aagtttacta cgattagaca aagcaaaaag gactggtaca 360

agtaggttat ccggtggaga acgaaaaagg ttatcgatag ctttggaact attaaacaat 420

cctcctgttt tatttctgga cgagccaact acgggtttag atgatctgtc atgttcccag 480

tgtatttcat tactgaaaaa gatcgccgaa ggtggtcgaa ctgtgatttg ctctatacat 540

acaccaagtg ccaaaatatt ttcacaattt gacaacgtct acattttatc tgacggacaa 600

tgcgtgtatc aaggttacgg acccgaagta gtatcctatt taagtaaagt aggaattgaa 660

tgtcctaaaa cctataaccc agcagatttt attatagaag tttgctgtaa cgaatatggt 720

aatttccaag aacggatggt atctgtgata gacaatggtc gtaatattta tagaactaac 780

ggtaacttac cacaactaca agaagtcgaa cttgccaacg atagtctaga aaatcgaagc 840

agtatttcat caatatcaat aagagaagaa gtttacgcgg atttaagttc ttcagatttt 900

aatcacgagt cttcatggtt tagccaattc tggatcttaa ccacccggtt atggatgcag 960

atgtggaggg ataaaactta cttaataatg aggactgtcc tatatataat ccttgctctt 1020

ctaataggat cactatatta taaaatgggt caggatgggt ccaaaaccat ctttaatttc 1080

ggtttttatt actgctgcat tatatttttc atgtatattc caatgatgcc aattctactg 1140

caatttcctc aagaaattca gttagtgaag agagaacatt ttaacaaatg gtaccgactt 1200

agtgcatatt tctcagcatt gtctttttcg accgtgcctg tacaactttg tcttggagtt 1260

gtgtatgtat cgtgtgttta ccttttaact gaccaaccgt tagagtttcg gaggatgtca 1320

atgttctttt tcatttgcat tttgaccagt gtaatttccg aaagtcttgg acttctgatt 1380

tctgctcagt tgaaagttgt gaatgctgtg tttatgggac cagtatctgc tgttcctttc 1440

atgctactag ctgtctacgg attcggatct ggttacgaca ctattccatc tataataaaa 1500

ttcttcatgc actttagcta cctcaggtac tcgctcgaag ggttgattca cgccatgtta 1560

aaagatagag aaaagctaag ctgtccggat actgaagaat actgtatata cacagacctc 1620

aacttattcg taagggacat gggcatggaa aacactattt attgggttga cgttttggtg 1680

ctaattttta ttttaatcct cttcaggggt ggtagttatt atctattacg acaaagatta 1740

actcctaaca aaacctttag agctctacaa tatattggaa gattggttaa atcacagttt 1800

ggactagcgc gatga 1815

<210> 36

<211> 1856

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 36

aagagaaaaa gattttgaaa aatgtcagtg gaatgctacg ttccggtgaa ctatgcgcta 60

ttatgggacc atctggtgct ggaaaatcca ccctactcaa tatactcact ggatacaaaa 120

ccataggggt aaaaggccat gtgttaatga acggcagtga tagagatctt agtcaattcc 180

gaaaattatc ggcatatatt atgcaggata accaactaca tgctaatcta agagtggatg 240

aagctatggc tgtcgcagct gcacttaaaa ttggcgccaa aagtccaaaa gatcgagaag 300

atatcattaa cgaaatccta gacacacttg gactgctgga tcataagaaa acaatgacaa 360

gcgggttgtc ggggggccaa aagaagagat tgtcaatcgc cttagaatta gttagtaatc 420

ctccagttat gtttttcgat gaacctacta gtggattgga ttcttcatca tgttttcaat 480

gtatatcact gttgaaaacc ttagctaaag gtggaagaac tataatatgt actattcacc 540

aaccttcagc tagactgttt gaaatgtttg accaacttta tacgctcgct gatggtcaat 600

gtgtttacca aggttctacg acattcttgg tgcccttttt ggcaagtctg cagttacagt 660

gtccttcata ccacaatcca gcctcctata ttatagaagt agcatgtggg gaatatggag 720

atcacacgag aacgttagtt aacgctatag aaaatggtaa gaatgacatc cgagatgcca 780

gccaaatagt aggtttcaaa atgagcgacg gtttgaataa cgcttaccaa tacgccaaag 840

acaacttgaa aacccttatt tctgatgaat tatccaaaga gggcagtacc gtgaatggca 900

atagaaaatc aaacaacgtt aacgaagata ctggaatcga ttctaaagat gtcgaaaaag 960

ctaatgttga tagtgcctta ttaaatactt ccgttgtggt taaacagccc agatatggta 1020

attcggaatt ccaacaattt ttcattattt taaaacgtgc gttattgttc agtagaagag 1080

attggacttt gatgtaccta cgtttattcg cccacatttt ggttggattc ctaattggtg 1140

ccttatactt caaaattgga aacgatgggt ccaaagtact tagtaatctt ggtttcctgt 1200

ttttcaacat gttgttcttg atgtacactt ccatgactat aaccattttg tccttcccac 1260

ttgaaatgcc agttttgtta aaagaacatt ttaatcgatg gtattccctg aggtcttact 1320

atctggcaat aacgatatca gatatgccat tccagaccat attctgtata ctttacgtaa 1380

cgatagtata tttcatgacg tcccaaccct tggatccggc tcgatttgga atgtttttac 1440

tgtcttcctt gttagtatca ttcgtggcac aaagtgttgg actcgtagta ggagctgcta 1500

tgaatgttca gaacggtgta ttcttagctc cagtaatgtc agtaccattc cttttgttct 1560

ccggattctt tgtatcattc gatgctattc ccatctacct tcgctggata acgtatctgt 1620

cctacatcag atacggattc gaagggactg cgcttgccac gtacggatac aacagaccaa 1680

acctggattg tttcgctcaa tattgccact ttaaaaaggc acttacaact ctcgaagagt 1740

tggacatgga taaatcgagt tactggttcg acattttggc tctagttgtg atattcttct 1800

ttttaagagt ttccgcttac ctcttcttga agtggaaact taaatcaaac cattaa 1856

<210> 37

<211> 1715

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 37

ctacaaaaga aattttgcat caagtcggag gacgtttcca gcccgggcag ctgatagcca 60

taatggggcc atcgggtgca ggcaaaagta cattattgga tgttttgagt gggtatagga 120

tacgtggcgt aacaggttca gtatacgtta acggccaacc aagaagttta aaggaattca 180

ggaaaacgtc atgttacatt acgcaagatg accgtctgca gcctttactc actgtggacg 240

aaaatatgtg ggtagctgca gatttgaaac taccttccag tgtcagcaac agtaataaac 300

ggagtataat taacaatatt ttaaaaacac taaatcttga aaatacaaaa aaagtgagag 360

cagctggtct ttccggtgga cagaaaaaga ggttgtcaat agctttagag ttagtaaata 420

atccaatggt catgttcttg gatgaaccaa ccacgggatt ggatagttcg tcttgttcaa 480

catgtataac actacttaag aaactaacta tgcaaggaaa aacaatcata tgtaccattc 540

atcagccttc agctacttta tttgcaatgt ttgatcaggt gtacgtagta ggaaatgggt 600

actgccttta tcaaggtagt acgaataaac tggttccatt tttgcaagac tgtgggtttc 660

cctgtccgca gtatcacaat cctgctgact acgtaataga actagcatgt attgagcacg 720

gacctgagaa acctgaatat atgagggaaa gaattcagaa cggacaatct taccaatatt 780

ttaacgagcc tgaaaagatg aaaagcacca tttgtaacta cccaggtccc aaacctatta 840

aacgagttgc tagcagtgat ggatcctctt ttcccacatc acagttccat caactaggag 900

ttcttatgag acggggattt attaaagcct acagagataa gatgcttacc taccttcgta 960

ttggaacaaa cataatagtt ggactgatgt tgggcagcct ctattggaaa gctggaagtg 1020

atggatctaa agttttggat aattttaatt tattgtttgc gatattattg caccatatga 1080

tgtccacaat gatgttgacg attttaacat ttccccagga aatgtcgatt ttaataaaag 1140

aacacttcaa ccgttggtat tcattaaaaa tgtattacac atcggtaacc atagtggata 1200

ttccggtttc ggttattggc tgcttcctgt tttcactcat aatatactat atgacagatc 1260

aaccaaacga taaagccaga ttcctgatat tcttcgtcac ttgcatgttg gtagttctag 1320

tagctcaagg agttggtcta actattggcg catattttga tgttgtgaac ggaacttttg 1380

ttggaccaac cctgatggta cccatgatga tgttctccgg ttttggcgtt cgtctacgtg 1440

atctgccgac cataatgtac tacggcagtt ttacttctta cttaaggtac ggcttggaag 1500

gagtagtcca ggcggtgtac ggaatggaca ggggtattct tcaatgtccc gaagacaaat 1560

tttgcttcta caagtatccg aagactttcc tggaaatagt agatgtcagg tcggatcagt 1620

tcgataatga catcattgct ttactgcttt tcttgtttgt tttgagaatt gcggcgtacg 1680

tagtgctgag gtacaaactg gcatctgtgc gatag 1715

<210> 38

<211> 1521

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 38

atgagtgaag ccagtggtag tgcaatggag gttatgttgc cgaaatttaa acctttaact 60

cagttaaata caatcgctaa aaaaccaccc attgatgttg aattctgtga tttaacttac 120

tcgatttctg attccagtta taaaggggga tggaggcaat tattaaaatc gatcaatgga 180

aagtttagat ctggagaact aactgccatt cttggtccct ctggcgcagg aaaatcaacg 240

ttacttaata ttttggcggg atatatgacg gcgggagtta aaggtagtgt aaaaatcaac 300

ggaaagcctc gagatatgag aatcttcacc aagctctcgt cgtacatcat gcaagaagat 360

ctagtgcaac cgaggttgag tgtcagagaa tcaatgatgg tagcagctaa tcttaaattg 420

agcgcgtcga tagggcatac acaaaaagtg gctgtggtac atgaagtgat ccaacttctt 480

ggtctagaaa aatgttacga taccaaaact gaataccttt ccggtggaca aagaaaacga 540

ctgagtgtag ctttagaatt agttaataac ccccctgtaa tatttttaga tgagcccaca 600

acgggattgg ataatgtatc tattaaacaa tgtatagatc ttcttaagaa aataactagg 660

ctcgaaagaa ctgtgatatg tacaatccat cagccgccag catcgctctt ccaaattttt 720

gatcaagtgt atattatggc taatggatat tgtgtttata acgggtcgcc aaatcaatta 780

gtacctttta tgtcgtcggt aaattgtgta tgtccagaaa cctcaacacc agctgatttt 840

attatagaag taatccaaac caatcaagac aatattccaa tactgcaaaa ccaaattcaa 900

aacggtaaaa taaatatgaa agataaaaag ttaaaacctc tacaaagtca caagacatta 960

ggaatatatg aaatttacca ggaaaccaca caaacaggta tgcatattca cgacatagaa 1020

tacccaacgt ctttctggac acaattcacc gtgcttttgt gtcgcatggc tttacagatg 1080

aaaagaaata aatccatgtg gatcatacaa tttttccatc acgtcatttc agctacgtta 1140

gtcggtggaa tattttacca gattggaaat gaagccagtc aagttctacc aatatttaag 1200

tattgtgtaa cgattaacgt gtttttcgtg tacactcatg tcatggtgcc agtgttactg 1260

tgtatttcga tagaagtcaa actgttaaaa cgagagtatt ttaatcgatg gtttagctta 1320

aagccgtact ttcttgctag tacgatagta aacattccaa tgttggttgg ttatggaatg 1380

atattcatat caatagtatt tttcatgacg ggtcaaccaa tagaatggca acgattctta 1440

atgtttacta tgatcgccat caacgtcgga ttttgttctc agggattagg atatgcaatt 1500

ggatccaatt gtggtatact g 1521

<210> 39

<211> 2128

<212> DNA

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 39

atgactcctg taggcccagg tgccaagacg aaaattgagc tggaacgctc ccatagcttg 60

ctcccaatta gacatattcc caatggtgac ttggccgtgg ctgcagtggt ttccctaggt 120

ggtcaaaggg ctacggtttc attaccaagt accaaaccaa cccacttaat tagcagccac 180

aaaccaagtg acgaacgaga gacgtttctc cgccaaggtg aacgtcaaat tctctcagaa 240

aacgaaatca ttctcgatag gtggagtacg ttccatcacc gagtggccct cgtccgcgtc 300

ccctttaacc ccgtcccagt ggttgtagac acgtgtaaac acagcgttcc agtcgctggc 360

gctgtcttac ctgtggtttc agtggcagta ttaggatttt taataagagt gccaaacgaa 420

tttgacatat atgcctcagc tagtttggcg aagaaaaggc aacgtaccta ctcacgatgg 480

tcacctattg aagagggagt aactttagtt tgggaaaatc tttcagtata ctcgtcaacg 540

acgaaaaatg gccagatgca gcacaaacag ataattaacg gagttactgg agctgttaaa 600

gcaggcagtt tggtagctat aatgggatca agcggtgctg gcaaaagtac tttgatgact 660

gctttaggat atagaactga aggttcaata ttaactgaag gcaacattct aataaacggt 720

cgacaaattg gggactatat gaaatattta agcggattta tgcaccaaga ggatatgttg 780

ctatcttatc ttacagtaag ggaacacatg aatataatgg ccaatttaaa gcttgataga 840

agactatcgg gaaatgataa aaaacagcta atatacgaca ttctcagaca acttggacta 900

atgaagtgca ttgatcttaa aattggaggt atagatcagg ccaaatcatt atctggtgga 960

gagaagaagc gattggcatt tgccacagag ttgcttacag atccacttct tttattctgt 1020

gacgaaccta caactggtct agactcgtat tctgctcaga aattggtggt aattatgaac 1080

caaatggcca ttaccggaaa aacgattttg tgtaccatac atcaaccttc ttcagatata 1140

tttgccatgt ttagtcaact aattttagtt gctgatggac gaatagctta tatgggttct 1200

acaaacaatg ctcttgactt ctttgagagg atgggttatg tatgtccaac atcttacaac 1260

cctgctgatt tttatattaa gacgttatcc acaactccgg gttatgaaga taactgtaga 1320

caaacagtta aaagaatatg cgaccaattt gctgttagtg atgaagcaaa agaagtcgaa 1380

atagtagtac aatatgaact tcatatgggc agagttgcaa cacaaagaaa atttgaattg 1440

cgtcaaaatt tcaaagagat tcgttggctg tcaaaattgt tttggttaac gtacagatgg 1500

attttggaaa tttatagaaa cccgtcattg gaggcaatga aaattgcaca aagaatgctg 1560

attggtttca tagtcgggtt ctgttatctt gggacagatg ccttgacaca gaatggtgta 1620

cagtctgtga caggaataat ctttatgttt gttagcgaaa atacatttaa tcccatgtac 1680

agtgtactgc atcaatttcc ttcttactta cccctatttt taagggaata caagtctggg 1740

ttatatcatc ctgcgactta ctatttatca agaattttat cactgctgcc aggattcata 1800

ttggaaccat taatatttgt aataactgct tactggttag ctggactacg tactagtgga 1860

tacgcatttc taatgacatt atcggtcatt gttctgacta tgaacgtttc atccgcgtgt 1920

ggtgtcatgt tttcaaatgc ttttgattct gttcctacgg ctttagccta tttggtaccc 1980

tttgactacg cccatatgat tacgtctggt gtcttcgtaa agttgagtac ccttccactt 2040

gtattttctt ggataaaata tctctcctgg ttaatgtatt cgacggaggc gttgtacata 2100

ttgcaatgga aagatgttac taatataa 2128

<210> 40

<211> 1160

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 40

Met Glu Thr Met Val Asn Leu Lys Arg Leu Thr His Ile Phe Asn Thr

1 5 10 15

Thr Ala Lys Lys Tyr Asn Met Ile Ile Ser Ala Glu Lys Thr Lys Cys

20 25 30

Met Thr Thr Ser Lys Tyr Pro Leu Arg Cys Lys Ile Glu Ile Asp Gly

35 40 45

Lys Ile Leu Lys Gln Glu Ala Arg Phe Arg Tyr Leu Gly Ile Asp Ile

50 55 60

Thr Ser Tyr Gly Asp Val Glu Gln Glu Val Arg Gln Gln Ser Leu Lys

65 70 75 80

Ala Ser Lys Gly Glu Gly Ser Leu Asn Asp Thr Ile Trp Lys Asn Glu

85 90 95

His Gln Arg Gln Asp Thr Lys Ala Ala Ile Arg Pro Ile Leu Thr Tyr

100 105 110

Thr Ala Lys Thr Arg Pro Asp Thr Ser Lys Thr Arg Arg Leu Leu Glu

115 120 125

Thr Thr Glu Met Lys Ile Leu Arg Arg Ile Ser Gly Lys Ser Leu Leu

130 135 140

Asp Arg Glu Ile Ser Glu Asn Ile Arg Arg Ser Cys Asn Val Glu Asp

145 150 155 160

Ile Asn Gly Trp Val Thr Lys Arg Lys Gln Glu Trp Asn Asp His Ile

165 170 175

Ser Arg Met Val Glu Asp Arg Ile Val Ser Leu Val Thr Phe Ala Thr

180 185 190

Tyr Val Leu Val Asp Glu Lys His Val Leu Asp Ala Asn Lys Ala Tyr

195 200 205

Val Ser Ile Ser Leu Phe Asn Ile Ile Arg Phe Pro Leu Ser Met Leu

210 215 220

Pro Met Met Leu Ser Asn Leu Val Gln Thr Trp Val Ser Val Lys Arg

225 230 235 240

Ile Asn Lys Phe Met Asn Ala Glu Glu Leu Asp Pro Asp Asn Val Leu

245 250 255

His Glu Pro Ser Glu Glu Lys Arg Arg Ala Thr Tyr Asp Arg Asn Gly

260 265 270

Thr Phe Ser Trp Gly Glu Gly Pro Ile Leu Lys Asp Ile Asn Ile Asn

275 280 285

Ile Lys Lys Lys Thr Leu Thr Ala Ile Val Gly Thr Val Gly Ser Gly

290 295 300

Lys Ser Ser Leu Ile Ser Ala Phe Leu Gly Glu Met Asp Lys Leu Ser

305 310 315 320

Gly Arg Val Asn Ser Tyr Gly Lys Val Ala Tyr Val Ser Gln Gln Ala

325 330 335

Trp Ile Gln Asn Ala Thr Leu Arg Asp Asn Ile Leu Phe Gly Lys Pro

340 345 350

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

355 360 365

Pro Asp Leu Glu Met Leu Pro Ala Gly Asp Gln Thr Glu Ile Gly Glu

370 375 380

Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg Val Ser Val Ala

385 390 395 400

Arg Ala Val Tyr Ser Asn Ala Asp Ile Tyr Phe Leu Asp Asp Pro Leu

405 410 415

Ser Ala Val Asp Ser His Val Gly Lys His Ile Phe Glu Lys Val Ile

420 425 430

Gly Pro His Gly Leu Leu Lys Ser Lys Thr Lys Ile Leu Val Thr His

435 440 445

Gly Ile Thr Tyr Leu Pro Gln Thr Glu Lys Ile Ile Val Leu Lys Asp

450 455 460

Gly Arg Ile Ser Glu Thr Gly Thr Tyr Gln Glu Leu Leu Asp Arg Lys

465 470 475 480

Gly Ala Phe Ala Glu Phe Leu Leu Gln His Ile Thr Glu Glu Ala Glu

485 490 495

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

500 505 510

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

515 520 525

Val Arg His Arg Ser Arg Val Ser Glu Ser Gln Ser Glu Thr Gly Ser

530 535 540

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

545 550 555 560

Ser Asn His Lys Leu Ser Ile Asp Asp Gly Lys Lys Gly Pro Lys Lys

565 570 575

Gly Glu Lys Leu Ile Glu Thr Glu Lys Ala Glu Thr Gly Asn Val Ser

580 585 590

Trp Ala Val Tyr Lys His Tyr Leu Lys Ser Ile Gly Leu Val Phe Met

595 600 605

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

610 615 620

Ser Asn Val Trp Leu Gly Leu Trp Ser Asp Asp Gln Lys Ile Val Val

625 630 635 640

Asn Asp Thr Val Asp Thr Ala Arg Arg Asp Met Tyr Leu Gly Val Tyr

645 650 655

Gly Ala Leu Gly Leu Gly Gln Val Val Thr Ile Leu Phe Ala Ser Leu

660 665 670

Ala Leu Tyr Val Gly Ser Leu Asn Ser Ala Arg Ala Leu His Asn Leu

675 680 685

Ile Leu Gly Asn Val Leu Lys Ala Pro Cys Thr Thr Phe Phe Asp Val

690 695 700

Thr Pro Leu Gly Arg Ile Leu Asn Arg Phe Ser Lys Asp Ile Asp Thr

705 710 715 720

Leu Asp Asn Val Leu Pro Met Thr Met Arg Gly Trp Ile Thr Cys Phe

725 730 735

Tyr Ser Val Val Gly Thr Leu Ala Val Ile Ser Tyr Ser Thr Pro Val

740 745 750

Phe Ile Leu Val Ile Ile Pro Leu Cys Phe Ile Tyr Tyr Phe Met Gln

755 760 765

Arg Phe Tyr Val Ala Thr Ser Arg Gln Val Lys Arg Leu Glu Ser Val

770 775 780

Ser Arg Ser Pro Ile Tyr Ser His Phe Gly Glu Ser Val Ser Gly Ala

785 790 795 800

His Ala Ile Arg Ala Tyr Asn Gln Gln Glu Arg Phe Ile Val Glu Ser

805 810 815

Glu Asn Lys Val Asp Met Asn Gln Ile Cys Tyr Tyr Pro Gly Ile Ile

820 825 830

Ser Asn Arg Trp Leu Ala Val Arg Leu Glu Met Ile Gly Asn Leu Ile

835 840 845

Ile Phe Phe Ala Ala Leu Phe Ala Val Leu Lys Lys Asp Gln Ala Pro

850 855 860

Gly Leu Val Gly Leu Ser Val Thr Tyr Ser Leu Gln Ile Thr Gln Thr

865 870 875 880

Leu Asn Trp Leu Val Arg Met Thr Ser Asp Val Glu Thr Asn Ile Val

885 890 895

Ala Val Glu Arg Ile Lys Glu Tyr Gly Glu Ala Pro Gln Glu Ala Ala

900 905 910

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

915 920 925

Thr Val Gln Phe Lys Asn Tyr Ser Val Arg Tyr Arg Pro Gly Leu Asp

930 935 940

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

945 950 955 960

Gly Ile Val Gly Arg Thr Gly Ala Gly Lys Ser Ser Leu Thr Leu Ala

965 970 975

Leu Phe Arg Ile Ile Glu Ala Ala Gln Gly Asn Ile Leu Ile Asp Gly

980 985 990

Ile Arg Ile Asp Glu Leu Gly Leu His Thr Leu Arg Ser Arg Leu Thr

995 1000 1005

Ile Ile Pro Gln Asp Ala Val Leu Phe Ser Gly Thr Leu Arg Ile

1010 1015 1020

Asn Leu Asp Pro Phe Asn Lys His Ser Asp Glu Asp Val Trp Arg

1025 1030 1035

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

1040 1045 1050

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

1055 1060 1065

Val Gly Gln Arg Gln Leu Ile Cys Leu Ala Arg Ala Leu Leu Arg

1070 1075 1080

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

1085 1090 1095

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

1100 1105 1110

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

1115 1120 1125

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

1130 1135 1140

Glu Phe Asp Ser Pro Ala Arg Leu Leu Ala Asn Glu Asn Ser Ile

1145 1150 1155

Phe Ala

1160

<210> 41

<211> 859

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 41

Met Glu Asn Leu Leu Trp Leu His His Pro Arg Pro Leu Lys Met Gln

1 5 10 15

Ala Ile Arg Ile Leu Arg Leu Tyr Glu Asp Glu Gly Ile Leu Gln Phe

20 25 30

Ile Ile His Ile Pro Ser Ala Gln Ala Asp Asn Thr Arg Cys Thr Val

35 40 45

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

50 55 60

Arg Ser Ile Ile Leu Val Arg Ser Glu Asn Glu Glu Lys Lys Asn Leu

65 70 75 80

Glu Lys Ser Met Cys Phe Val Ser Arg Gly Lys Leu Gln Arg Tyr Cys

85 90 95

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

100 105 110

Met His Ala Tyr Asn Thr Ile Met Thr Lys Thr Pro Asn Lys Glu Asn

115 120 125

Ala Ser Phe Leu Val Cys Ala Val Gln Ala Ile Ser Trp Cys Ser His

130 135 140

Phe Leu Tyr Thr Val Gly Leu Arg Ser Arg Leu Gly Lys Ser Gln Arg

145 150 155 160

Gly Pro Thr Ala Met Gly Ile Val Trp Cys Met Val Phe Ala Met Thr

165 170 175

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

180 185 190

Glu Asn Thr Lys Met Glu Leu Gly Ile Thr Leu Tyr Tyr Thr Val Leu

195 200 205

Gln Ile Ile Tyr Ala Leu Ser Leu Ile Pro Gly Gly Ser Thr Thr Thr

210 215 220

Leu Asn Phe Pro Glu Arg Tyr Thr Glu Ile Thr Glu Arg Gln Pro Leu

225 230 235 240

Ile Asn Ala Asn Ala Tyr Gly Arg Phe Ser Glu Glu Gly Asp Pro Asn

245 250 255

Asp Leu Gly Val Ala Met Glu Asp Thr Asn Trp Leu Ser Arg Leu Ser

260 265 270

Phe Ser Trp Val Lys Ser Leu Val Asn Lys Gly Val Glu Asp Lys Ile

275 280 285

Val Thr Ser Asp Asp Leu Tyr Asp Leu Pro Asp Ser Ile Ser Ser Asp

290 295 300

Asn Asn Ser Asn Arg Leu Glu Asn Tyr Leu Arg Ile Glu His Ala Ala

305 310 315 320

Leu Cys Asp Ser His Phe Asn Phe Lys Met Ser Val Ile Gly Leu Lys

325 330 335

Met Arg Gly Ala Ile Ile Asn Thr Ile Tyr Arg Lys Thr Leu Ser Val

340 345 350

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

355 360 365

Tyr Met Ser Thr Asp Thr Asp Arg Ile Val Asn Ser Cys Pro Ser Phe

370 375 380

His Ala Val Trp Ser Ile Pro Phe Gln Leu Phe Val Ser Leu Tyr Leu

385 390 395 400

Leu Tyr Asn Gln Val Gly Leu Ala Phe Leu Ala Gly Val Leu Phe Ser

405 410 415

Ile Ile Leu Ile Pro Ile Asn Lys Cys Ile Ala Asn Lys Ile Gly Asp

420 425 430

Leu Ser Thr Lys Met Met Glu Gln Lys Asp Gly Arg Val Lys Leu Thr

435 440 445

Ser Glu Ile Leu Arg Gly Ile Lys Ala Ile Lys Leu Tyr Val Trp Glu

450 455 460

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

465 470 475 480

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

485 490 495

Gln Ser Glu Tyr Thr Ser Ser Glu Cys Thr Leu Glu Asn Ser Leu Cys

500 505 510

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

515 520 525

Val Ser Glu Arg Gly Asn Leu Asp Phe Thr Val Ile Ile Ser Tyr Trp

530 535 540

Lys Gly Ile Ser His Phe Val Ser Ile Ser Ile Leu Ile Ser Met Thr

545 550 555 560

Leu Met Gln Val Ser Arg Asn Phe Thr Asp Trp Trp Leu Ala Asn Gly

565 570 575

Val Thr Ser Pro Val Thr Asn Ser Thr Asn Leu Thr Val Tyr Leu Ala

580 585 590

Thr Ala Thr Asp Tyr Ile Asp Tyr Val Glu Asp Asp Asp Asn Met Ser

595 600 605

Glu Phe Leu Lys Val Tyr Val Glu Leu Ala Cys Val Asn Thr Leu Phe

610 615 620

Thr Leu Ile Arg Ser Phe Ile Phe Ala Tyr Gly Gly Ile Leu Ala Ala

625 630 635 640

Thr Lys Phe His Lys Val Leu Leu Lys Thr Val Leu Arg Ala Arg Cys

645 650 655

Thr Phe Phe Asp Val Thr Pro Ile Gly Arg Ile Ile Asn Arg Phe Ser

660 665 670

Ser Asp Thr Tyr Thr Val Asp Asp Ser Leu Pro Phe Ile Leu Asn Ile

675 680 685

Phe Leu Ala Gln Leu Phe Gly Leu Leu Gly Ser Leu Phe Ile Thr Met

690 695 700

Tyr Gly Leu Pro Trp Ile Cys Val Phe Leu Ile Pro Leu Val Pro Ile

705 710 715 720

Tyr Thr Tyr Leu Leu Asn Gln Tyr Arg Ile Thr Ser Arg Glu Leu Lys

725 730 735

Arg Ile Ser Ser Val Thr Leu Ser Pro Ile Tyr Asn His Phe Asn Glu

740 745 750

Thr Leu Gln Gly Leu Thr Thr Ile Arg Ala Met Arg Val Ile His Lys

755 760 765

Phe Lys His Asp Asn Asn Gln His Leu Glu Ala Asn Leu Lys Ala Gln

770 775 780

Phe Ala Ser Gln Ala Ala Ala Arg Trp Leu Gly Leu Arg Leu Gln Phe

785 790 795 800

Ile Gly Val Ile Ile Ile Thr Gly Val Ser Phe Ile Ala Val Ile Gln

805 810 815

His Gln Tyr Asp Ile Ala Asp Pro Gly Phe Ile Gly Leu Ala Ile Ser

820 825 830

Tyr Ala Leu Gly Ile Thr Gly Ser Leu Ser Gly Val Val Asn Ser Phe

835 840 845

Thr Glu Thr Glu Arg Glu Met Val Ala Val Glu

850 855

<210> 42

<211> 829

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 42

Asn Met Leu Asn Ile Ser Tyr Cys Pro Pro Asn Ile Ser Val Trp Asp

1 5 10 15

Ile Trp Val Asp His Gly Ile Pro Gln Cys Phe Met Asn Thr Val Thr

20 25 30

Ser Ser Val Leu Ala Ile Tyr Ile Leu Ile Ala Gly Ser Ile Gln Leu

35 40 45

His Ile Tyr Arg Arg Trp Gly Val Glu Ser Gly Thr Arg Phe Leu Pro

50 55 60

Arg Gly Arg Leu Tyr Tyr Leu Gln Thr Phe Leu Ile Leu Phe Ile Pro

65 70 75 80

Val Leu Glu Ile Val Arg Phe Ile Leu Thr Ala Thr Val Tyr Asp Asp

85 90 95

Lys His Ile Tyr Gly Tyr Met Ile Val Ser Leu Val Leu Thr Thr Phe

100 105 110

Ala Phe Pro Phe Ser Leu Trp Val Leu Lys Val Glu Leu Tyr Asn Ile

115 120 125

Leu Pro Ser Val Gln Thr Arg Gly His Gly Ile Val Leu Leu Leu Phe

130 135 140

Trp Thr Leu Ala Phe Ile Ser Glu Asn Leu Ala Phe Ile Asn Leu Thr

145 150 155 160

Gln Glu Asn Trp Trp Phe Lys Leu Lys Asp Leu Thr Asp Gln Leu Glu

165 170 175

Met Ala Leu Phe Ile Leu Arg Tyr Ile Ala Cys Leu Leu Ile Phe Phe

180 185 190

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

195 200 205

Leu Gly Gly His Gln Arg Asn Val Ile Asn Pro Asn Asn Glu Asn Ala

210 215 220

Ser Thr Trp Lys Asn Phe Trp Lys Lys Val Lys Ile Leu Ala Pro Phe

225 230 235 240

Leu Trp Pro Lys Lys Asn Phe Thr Leu Gln Phe Lys Val Leu Ile Cys

245 250 255

Phe Ile Leu Leu Leu Gly Gly Arg Ala Val Asn Leu Phe Val Pro Ile

260 265 270

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

275 280 285

Leu Phe Arg Trp Asp Trp Val Leu Ile Tyr Val Gly Leu Lys Phe Leu

290 295 300

Gln Gly Gly Gly Thr Gly Gly Met Gly Leu Leu Asn Asn Val Arg Ser

305 310 315 320

Phe Leu Trp Ile Arg Val Gln Gln Tyr Thr Thr Arg Glu Val Glu Val

325 330 335

Glu Leu Phe Arg His Leu His Ser Leu Ser Leu Lys Trp His Leu Gly

340 345 350

Arg Lys Thr Gly Glu Val Leu Arg Val Met Asp Arg Gly Thr Asp Ser

355 360 365

Ile Asn Asn Leu Leu Asn Tyr Ile Ile Phe Ser Ile Phe Pro Thr Ile

370 375 380

Val Asp Ile Ile Val Ala Ile Val Phe Phe Val Ser Ala Phe Asn Ile

385 390 395 400

Trp Phe Gly Leu Ile Val Phe Thr Thr Met Ile Leu Tyr Ile Val Leu

405 410 415

Thr Ile Val Ile Thr Glu Trp Arg Thr Lys Phe Gln Arg Arg Met Asn

420 425 430

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

435 440 445

Phe Glu Thr Val Lys Tyr Tyr Gly Ala Glu Asn Tyr Glu Val Asp Ala

450 455 460

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

465 470 475 480

Ile Thr Leu Asn Ile Leu Asn Thr Val Gln Asn Val Ile Ile Cys Gly

485 490 495

Gly Leu Leu Ala Gly Ser Leu Leu Cys Val Tyr Met Val Val Glu Pro

500 505 510

Lys Thr Leu Lys Ala Gly Asp Tyr Val Leu Phe Ala Thr Tyr Ile Val

515 520 525

Gln Leu Tyr Val Pro Leu Asn Trp Phe Gly Thr Tyr Tyr Arg Ala Ile

530 535 540

Gln Lys Asn Phe Val Asp Met Glu Asn Met Phe Asp Leu Leu Arg Glu

545 550 555 560

Glu Gln Glu Ile Ile Asp Ala Pro Gly Ala Thr Asn Leu Ser Val Pro

565 570 575

Arg Gly His Val Glu Phe Lys Asn Val Ser Phe Gly Tyr Leu Pro Glu

580 585 590

Lys Leu Ile Leu Lys Asn Ile Thr Phe Ser Val Pro Ser Gly Lys Thr

595 600 605

Val Ala Leu Val Gly Pro Ser Gly Ser Gly Lys Ser Thr Ile Ile Arg

610 615 620

Leu Leu Phe Arg Phe Tyr Asp Val Asp Thr Gly Val Ile Val Ile Asp

625 630 635 640

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

645 650 655

Gly Val Val Pro Gln Asp Thr Val Leu Phe Asn Asn Thr Val Glu Tyr

660 665 670

Asn Ile Gln Tyr Gly Arg Leu Thr Ala Thr Val Thr Asp Val Ile Glu

675 680 685

Ala Ala Arg Gly Ala Asp Ile His Glu Lys Ile Leu Thr Phe Pro Glu

690 695 700

Ala Tyr Glu Thr Lys Val Gly Glu Arg Gly Leu Arg Leu Ser Gly Gly

705 710 715 720

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

725 730 735

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

740 745 750

Arg Asn Ile Gln Glu Ser Leu Asn Arg Met Cys Ala Asn Lys Thr Thr

755 760 765

Ile Ile Val Ala His Arg Leu Ser Thr Ile Ile His Ala Asp Glu Ile

770 775 780

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

785 790 795 800

Leu Ile Gly Gln Glu Gly Ile Tyr Ala Asn Met Trp Arg Gln Gln Leu

805 810 815

Glu Asn Lys Asp Lys Glu Ser Leu Glu Asn Ser Ser Glu

820 825

<210> 43

<211> 1642

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 43

Met Ala Asp Asp Gly Lys Ser Leu Phe Phe Ser Gln Leu Lys Ala Met

1 5 10 15

Leu Lys Arg Asn Leu Leu Leu Lys Lys Arg Gln Lys Leu Lys Thr Ile

20 25 30

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

35 40 45

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

50 55 60

Gly Glu Glu Tyr Leu Phe Lys Ser Phe Asp Asn Gly Glu Cys His Lys

65 70 75 80

Ile Ala Tyr Ala Pro Asn Thr Thr Thr Asn Ile Asp Phe Val Asn Asn

85 90 95

Val Thr Glu Thr Trp Lys Lys Met Ser Asn Ser Lys Ser Ser Leu Glu

100 105 110

Trp Val Pro Tyr Ser Thr Glu Asp Lys Val Gln Glu Ala Tyr Asp Gln

115 120 125

Asp His Lys Phe Ala Pro Met Ala Val Ile Phe Lys Val Pro Tyr Val

130 135 140

His Thr Ser Leu Ser Tyr Gln Ile Arg Thr Asn Pro Tyr Tyr Ile Ser

145 150 155 160

Thr Pro Ser Ser Ser Asp Ile Gly Trp Asp Lys Gln Ser Cys Arg Gly

165 170 175

Gly Thr Asn Pro Asn Ser Asn Met Glu Asp Gly Ser Thr Cys Pro Ala

180 185 190

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

195 200 205

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

210 215 220

Ile Ser Leu Glu Ile Phe Pro Lys Ala Ala Thr Thr Val Gly Asp Phe

225 230 235 240

Ala Asp Met Ile Arg Ile Ile Ile Pro Ile Tyr Met Val Leu Ala Leu

245 250 255

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

260 265 270

Lys Lys Ile Lys Glu Gly Met Lys Leu Met Gly Leu Met Asp Lys Val

275 280 285

Phe Trp Leu Ser Trp Leu Ile Ile Tyr Ala Cys Phe Val Ile Leu Leu

290 295 300

Ala Thr Ile Cys Val Leu Leu Leu Phe Ala Phe Lys Val Phe Val His

305 310 315 320

Val Asn Leu Leu Leu Ile Phe Ile Leu Met Val Leu Tyr Gly Ile Ser

325 330 335

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

340 345 350

Thr Ala Gly Val Leu Ser Ser Phe Ile Leu Val Leu Ile Ile Val Leu

355 360 365

Tyr Phe Val Glu Gln Ile Val Ser Gly Lys His Pro Gln Tyr Leu Trp

370 375 380

Ala Leu Ser Leu Leu Ser Pro Ala Gly Phe Ala Ser Ala Leu Asp Lys

385 390 395 400

Val Ala Ile Ser Asp Ile Gln Gly Lys Gly Leu Asp Phe Ser Asn Met

405 410 415

Trp Asp Asn His Gly Ser Gly Val Pro Phe Gly Gly Ser Leu Val Met

420 425 430

Ile Met Val Asp Ile Val Leu Tyr Ala Ile Ile Gly Trp Tyr Leu Asp

435 440 445

Asn Val Ile Pro Ser Ser Tyr Gly Val Lys Arg Ser Pro Ile Phe Phe

450 455 460

Leu Met Pro Ser Tyr Trp Arg Ser Ser His Asn Gln Ile Pro Pro Asn

465 470 475 480

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

485 490 495

Met Gln Asp Lys Glu Ala Ile Arg Ile Ile Asn Leu Asn Lys Ser Phe

500 505 510

Thr Gln Cys Arg Lys Pro Thr Val Thr Ala Leu Asp Gly Ile Asn Leu

515 520 525

Ser Ile Tyr Lys Gly Gln Ile Thr Ala Ile Leu Gly His Asn Gly Ala

530 535 540

Gly Lys Thr Thr Leu Phe Asn Ile Leu Thr Gly Leu Ser Ser Pro Thr

545 550 555 560

Ser Gly Ser Ala Leu Val Phe Gly Tyr Asp Val Ser Asn Pro Asn Asp

565 570 575

Met Asp Lys Ile Arg Arg Met Thr Gly Val Cys Pro Gln His Asp Ile

580 585 590

Leu Phe Asp Asp Leu Thr Pro Arg Glu His Leu Glu Phe Phe Ala Ala

595 600 605

Ile Lys Gly Ile Ser Asn Arg Gln Ser Ala Ile Glu Lys Ile Ile Arg

610 615 620

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

625 630 635 640

Gly Gly Gln Lys Arg Lys Leu Ser Ile Gly Ile Ala Leu Ile Gly Asp

645 650 655

Pro Lys Ile Ile Ile Leu Asp Glu Pro Thr Ala Gly Val Asp Pro Tyr

660 665 670

Ser Arg Arg His Leu Trp Asn Val Leu Gln Asn Val Arg Arg Asp Lys

675 680 685

Val Ile Leu Leu Thr Thr His Phe Met Asp Glu Ala Asp Ile Leu Ala

690 695 700

Asp Arg Lys Ala Val Val Ser Lys Gly Lys Ile Arg Cys Cys Gly Ser

705 710 715 720

Ser Leu Phe Leu Lys Asn Lys Phe Gly Ile Gly Tyr His Leu Thr Phe

725 730 735

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

740 745 750

Gln Phe Val Lys Met Ala Arg Lys Asp Arg Arg His Gly Lys Glu Leu

755 760 765

Ser Phe Ile Leu Pro His Asn Ala Val Glu Asn Phe Ala Ser Leu Phe

770 775 780

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

785 790 795 800

Ser Tyr Gly Val Ser Met Thr Thr Leu Glu Glu Val Phe Leu Gly Leu

805 810 815

Gln Lys Glu Glu Glu Tyr Gly Asp Ala Thr Ala Tyr Gln Asp Leu Pro

820 825 830

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

835 840 845

Ile Arg Ser Leu Glu Ala Phe Arg Cys Thr Pro Ser Thr Ser Gln Asn

850 855 860

Leu Arg Thr Leu Ile Gly Leu Arg Leu Leu Arg Leu Arg Arg Glu Lys

865 870 875 880

Arg Lys Leu Trp Met Val Ile Ile Leu Pro Ile Ile Phe Thr Gly Leu

885 890 895

Gly Leu Tyr Leu Asn Lys Val Ile Asp Thr Gly Lys Lys Tyr Glu Pro

900 905 910

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

915 920 925

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

930 935 940

Asn Gln Leu Glu Lys Asn Gly Ala Glu Leu Asp Thr Tyr Asp Gly Glu

945 950 955 960

Phe Thr Ser Leu Leu Asp Ile Ala Pro His Tyr Ala Ser Phe Asn Ile

965 970 975

His Asp Phe Ser Ser Ser Ser Gln Asn Ile Ser Val Leu Tyr Asn Gly

980 985 990

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

995 1000 1005

Phe Thr Val Ala Met Arg Ser Met Ala Ala Thr Thr Gly Lys Ile

1010 1015 1020

Ala Val Thr Thr Tyr Pro Phe Glu Leu Pro Pro Thr Asn Val Ser

1025 1030 1035

Pro Thr Pro Ala Ser Pro Gly Asn Phe Ile Leu Gly Met Ile Phe

1040 1045 1050

Leu Phe Ala Pro Ile Val Leu Ala Val Asp Met Val Tyr Glu Arg

1055 1060 1065

Glu Ile Lys Ala Arg Asn Gln Leu Arg Val Asn Gly Leu Pro Phe

1070 1075 1080

Thr Val Tyr Phe Thr Ser Phe Phe Leu Val Gln Ile Leu Leu Met

1085 1090 1095

Ala Ile Ile Thr Ile Leu Leu Ile Ile Leu Ile Val Val Met Lys

1100 1105 1110

Pro Pro Thr Phe Ser Asn Ser Ser Ile Thr Val Leu Ala Ile Trp

1115 1120 1125

Val Ile Leu Tyr Cys Pro Ala Ser Val Leu Phe Cys Ser Cys Val

1130 1135 1140

Ser Tyr Met Phe Asp Lys Ser Glu Ser Ala Gln Ser Val Met Pro

1145 1150 1155

Asn Val Ser Thr Leu Leu Gly Leu Ile Pro Tyr Ile Ser Val Met

1160 1165 1170

Tyr Ala Lys Glu Ser Val Ala Thr Ile Leu His Tyr Val Phe Ala

1175 1180 1185

Phe Thr Asp Met Met Tyr Val Pro Tyr Gly Met Leu Tyr Tyr Ile

1190 1195 1200

Gln Lys Ile Asn Leu Glu Cys Ile Ser Asp Pro His Cys Glu Gly

1205 1210 1215

Thr Thr Phe Ser Ser Phe Met Ile Pro Glu Ile Ile Val Leu Phe

1220 1225 1230

Val Thr Leu Ile Ile Gln Ile Pro Leu Leu Phe Leu Ile Met Leu

1235 1240 1245

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

1250 1255 1260

Arg Ser Lys Lys Ile Ser Glu Asn Ile Val Glu Glu Ser Arg Asp

1265 1270 1275

Val Gly Val His Glu Asp Arg Asp Val Lys Asn Glu Arg Gln Arg

1280 1285 1290

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

1295 1300 1305

Thr Val Glu Asn Leu His Lys Val Tyr Gln Lys Gly Val Lys Ser

1310 1315 1320

Gly Ile Cys Ser Arg Ser Ala Glu Ser Leu Lys Val Ala Ile Lys

1325 1330 1335

Ser Ile Ser Leu Ala Val Asp Ser Ala Glu Val Phe Gly Leu Leu

1340 1345 1350

Gly His Asn Gly Ala Gly Lys Thr Thr Ala Met Lys Ile Ile Thr

1355 1360 1365

Ala Glu Glu Ala Pro Thr Arg Gly Arg Val Gln Ile Val Gly Arg

1370 1375 1380

Asn Ile Thr Ser Ser Ala Asn Ala Ala Phe Gln Tyr Leu Gly Tyr

1385 1390 1395

Cys Pro Gln His Asp Ala Gln Trp Lys Asn Ile Thr Val Lys Glu

1400 1405 1410

His Leu Glu Leu Tyr Ser His Ile Arg Gly Ile Pro Lys Asn Glu

1415 1420 1425

Ile Lys Arg Thr Val Asp Leu Tyr Leu Thr Gly Leu Gln Ile His

1430 1435 1440

Glu His Lys Asp Lys Gln Ala Glu Lys Cys Ser Gly Gly Thr Arg

1445 1450 1455

Arg Lys Leu Ser Tyr Ala Met Ala Met Ile Gly Asn Pro Lys Ile

1460 1465 1470

Val Leu Leu Asp Glu Pro Ser Thr Gly Met Asp Pro Gln Ser Lys

1475 1480 1485

Arg Phe Leu Trp Asp Thr Val Leu Ala Ser Phe Arg Gly Ser Arg

1490 1495 1500

Gly Ala Ile Leu Thr Thr His Ser Met Glu Glu Ala Asp Ala Leu

1505 1510 1515

Cys Ser Arg Ile Gly Ile Met Val Asn Gly Glu Leu Arg Cys Leu

1520 1525 1530

Gly Ser Ser Gln His Leu Lys Asn Leu Tyr Gly Ala Gly Tyr Asn

1535 1540 1545

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

1550 1555 1560

Lys Leu Lys Glu Leu Glu Asn Tyr Val Phe His Leu Phe Pro Asn

1565 1570 1575

Ala Val Leu Gln Glu Thr Phe Gly Asp Arg Leu Ile Phe Ser Val

1580 1585 1590

Pro Gln Gln Ser Val Pro Ser Leu Ala Asn Cys Phe Arg Ser Leu

1595 1600 1605

Glu Asp Gly Lys Arg Arg Leu Asn Ile Glu Glu Tyr Ser Phe Ser

1610 1615 1620

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

1625 1630 1635

Glu His Glu Asp

1640

<210> 44

<211> 1387

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 44

Glu Leu Met Lys Met Val Gly Met Lys Ser Trp Met Leu Trp Phe Gly

1 5 10 15

Trp Phe Ile Tyr Ala Met Ile Pro Met Phe Phe Ala Val Ser Val Ile

20 25 30

Ser Ile Phe Met Lys Val Pro Met Phe Gly Ser Asp Ser Pro Leu Val

35 40 45

Glu Phe Ala Asn Gly Gly Ile Leu Phe Val Phe Leu Ile Leu Tyr Cys

50 55 60

Met Ala Ala Val Ala Phe Cys Phe Ala Ile Ser Ser Phe Phe Ser Arg

65 70 75 80

Pro Thr Ile Ala Leu Val Ala Gly Ile Leu Val Trp Ile Leu Ser Phe

85 90 95

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

100 105 110

Trp Leu Ser Asn Val Leu Leu Asn Leu Leu Pro Asn Met Ser Leu His

115 120 125

Tyr Gly Tyr Ser Ile Ile Ser Val Phe Glu Glu Arg Glu Val Gly Ile

130 135 140

Asn Trp Ser Asn Phe Phe Lys Pro Gly Ser Gly Ser Ser Asp Asp Ile

145 150 155 160

Thr Met Leu Asn Val Tyr Val Met Leu Ile Val Asp Ile Val Ile Tyr

165 170 175

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

180 185 190

Val Arg Lys Ser Ile Leu Phe Pro Leu Gln Asn Phe Met Lys Leu Cys

195 200 205

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

210 215 220

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

225 230 235 240

Asn His Leu His Lys Arg Tyr Lys Gln Lys Gln Ala Val Asn Asn Leu

245 250 255

Asn Leu Asp Ile Tyr Lys Asn Gln Ile Thr Val Leu Leu Gly His Asn

260 265 270

Gly Ala Gly Lys Ser Thr Thr Met Ser Ile Ile Thr Gly Leu Ile Pro

275 280 285

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

290 295 300

Met Asp Glu Ile Arg Lys Ser Leu Gly Leu Cys Pro Gln His Asn Leu

305 310 315 320

Leu Phe Thr Asp Leu Thr Val Lys Glu His Leu Leu Phe Phe Ala Lys

325 330 335

Leu Lys Gly Lys Ser Thr Lys Glu Ala Asn Ile Glu Ala Lys Ser Leu

340 345 350

Leu Ala Lys Leu Asn Met Pro Asp Lys Glu His Ser Met Ala Gln Thr

355 360 365

Leu Ser Gly Gly Met Gln Arg Lys Leu Cys Leu Ala Met Ala Leu Ile

370 375 380

Gly Asp Ser Lys Val Leu Ile Leu Asp Glu Pro Ser Ser Gly Met Asp

385 390 395 400

Pro Gln Ser Arg Arg Glu Leu Trp Asp Leu Leu Leu Gln Trp Arg Gly

405 410 415

Glu Lys Thr Ile Leu Ile Thr Thr His Phe Met Glu Glu Ala Asp Ala

420 425 430

Leu Gly Asp Trp Ile Ala Ile Met Ser Asp Gly Ser Leu Asn Cys Tyr

435 440 445

Gly Thr Pro Met Phe Leu Lys Lys Lys Tyr Asp Thr Gly Tyr His Leu

450 455 460

Asn Leu Met Ile Glu Glu Asp Ala Asp Ile Asp Ala Ile Ser Arg Arg

465 470 475 480

Val Lys His Phe Met Pro Asp Ala His Leu Thr Ser Asn Asn Gly Asn

485 490 495

Asn Leu Val Phe Val Leu Pro Tyr Gln Asn Thr Asn Met Thr Gly Leu

500 505 510

Leu Gly Asp Leu Glu Lys Asn Arg Ser Glu Leu His Leu Ser Asn Ile

515 520 525

Ser Ile Thr Ile Thr Thr Leu Glu Asp Val Phe Leu Lys Thr Arg Gln

530 535 540

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

545 550 555 560

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

565 570 575

Lys Val Asn Phe Ser Leu Lys Lys Trp Ser Thr Tyr Ile Val Pro Val

580 585 590

Gly Val Ala Val Val Leu Phe Ser Leu Thr Val Phe Leu Ser Ser Gly

595 600 605

Asn Asp Tyr Tyr Ser Glu Arg Gly Pro Glu Leu Pro Leu Lys Leu Ser

610 615 620

Ser Tyr Lys Gln Thr Ser Val Tyr Tyr Ser Gly Asp Thr Ser Asn Glu

625 630 635 640

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

645 650 655

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

660 665 670

Arg Ser Leu Glu Asn Ile Ala Tyr Tyr Lys Glu His Met Ile Ala Gly

675 680 685

Ala Asn Phe Val Tyr Asp Arg Asp Phe Thr Leu Val Ala Leu Tyr Asn

690 695 700

Gly Ile Ala Thr His Ser Thr Pro Ile Ser Leu Asn Leu Ile Thr Asp

705 710 715 720

Ser Leu Ala Lys Thr Leu Leu Gly Pro Glu Tyr Gly Ile Ser Val Ser

725 730 735

Asn Trp Pro Leu Pro Gln Val Gln Glu Arg Leu Ser Ser Gln Glu Tyr

740 745 750

Ser Glu Ala Lys Val Ala Val Leu Trp Leu Ile Leu Leu Pro Val Gly

755 760 765

Cys Leu Phe Ile His Gly Ile Phe Ile Ile Phe Pro His Thr Glu Ile

770 775 780

Ser Thr Arg Phe Leu Gln Ile Gln Tyr Met Ala Gly Val Lys Pro Phe

785 790 795 800

Phe Tyr Trp Leu Val Asn Trp Ile Ala Asp Met Thr Phe Tyr Ile Phe

805 810 815

Leu Met Phe Ile Leu Ser Phe Leu Leu Trp Ile Cys Ser Pro Val Phe

820 825 830

Gln His Asn Gly Thr Glu Leu Gly Tyr Leu Phe Ser Ile Phe Leu Cys

835 840 845

Tyr Gly Ile Ala Gly Ile Pro Phe Ala Tyr Ile Phe Ser Arg Lys Lys

850 855 860

Thr Ala Ser Gly Ala Phe Ala Leu Phe Val Ile Met Gly Met Phe Leu

865 870 875 880

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

885 890 895

Glu Tyr Tyr Val Asn Ile Gly Asn Lys Ile Lys Tyr Val Cys Phe Phe

900 905 910

Leu Leu Pro Gln Val Ser Leu Ser Asp Ala Leu Val Ser Phe Ile Arg

915 920 925

Arg Thr Val Asn Ile Tyr Asn Phe Lys Ile Ser Pro Gln Arg Leu Arg

930 935 940

Val Met Cys Asn Gln Met Glu Ser His Ala Cys Cys Val Glu Ser Ser

945 950 955 960

Ile Glu Cys Gln Asn Tyr Lys Ser Tyr Asn Asn Leu Phe Ser Glu His

965 970 975

Tyr Met Phe Met Ile Gly Cys Gly Val Phe Tyr Leu Thr Ile Asn Ile

980 985 990

Ile Leu Asp Thr Tyr Phe Met Lys Lys Leu Lys Ala Met Val Met Arg

995 1000 1005

Gln Cys Asn Leu Ile Leu Lys Ser Phe Lys Asp Lys Asp Thr Ala

1010 1015 1020

Leu Ile Pro Lys Asn Asp Lys Tyr Val Asn Glu Asp Val Gly Asp

1025 1030 1035

Gly Tyr Asn Thr Leu Arg Ala Lys Lys Ile Met Lys Leu Tyr Ala

1040 1045 1050

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

1055 1060 1065

His Cys Leu Gly Ile Leu Gly Val Asn Gly Ala Gly Lys Thr Thr

1070 1075 1080

Thr Phe Lys Met Leu Thr Arg Glu Glu Val Val Asp Asp Gly Glu

1085 1090 1095

Ile Lys Ile Glu Leu Asp Asn Asn Lys Gln Pro Leu Asp Ile Thr

1100 1105 1110

Gly Ser Glu Tyr Leu Glu Thr Ile Gly Tyr Cys Pro Gln Ser Asp

1115 1120 1125

Ser Leu Asn Phe Val Leu Thr Gly Arg Gln Leu Leu Ser Thr Val

1130 1135 1140

Ala Lys Leu Arg Gly Val Thr Asp Glu Thr Met Ile Asp Arg Phe

1145 1150 1155

Leu Glu Ala Phe Asp Leu Lys Gln Tyr Ala Asp Ile Pro Cys Gly

1160 1165 1170

His Tyr Ser Gly Gly Asn Lys Arg Lys Leu Ser Leu Ala Ile Ser

1175 1180 1185

Leu Ile Gly Asn Arg Lys Phe Val Leu Leu Asp Glu Pro Thr Asn

1190 1195 1200

Gly Val Asp Pro Ala Thr Arg Arg Lys Cys Trp Asp Leu Ile Lys

1205 1210 1215

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

1220 1225 1230

Ser His Ser Met Thr Glu Cys Glu Ala Leu Cys Asp Glu Leu Lys

1235 1240 1245

Ile Met Lys Lys Gly Ser Phe Val Glu Glu Gly Arg Leu Val Asp

1250 1255 1260

Leu Lys Asn Arg Tyr Gly Gly Phe Thr Leu Lys Leu Lys Leu Val

1265 1270 1275

Ser Asn Gln Gly Gln Pro Ile Asn Leu Val Asp Asp Asp Val Asp

1280 1285 1290

Glu Val Asp Gly Val Ile Ser Asn Lys Phe Lys Ser Val Asp Asp

1295 1300 1305

Leu Lys Ser Tyr Phe Thr Asn His Asp Arg Gly Glu Ile Lys Asp

1310 1315 1320

Glu His Ser Gly Leu Leu His Val Tyr Ile Lys Asp Lys Thr Lys

1325 1330 1335

Lys Trp Ser Asp Ile Phe Gln Glu Val Glu Ala Leu Lys Thr His

1340 1345 1350

Asn Ser His Leu Ile Glu Asp Tyr Ala Ile Ser Glu Ala Ser Leu

1355 1360 1365

Glu Asp Val Phe Leu Lys Val Ala Arg Glu Asp Glu Glu Asp Val

1370 1375 1380

Thr Lys Lys Lys

1385

<210> 45

<211> 1362

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 45

Met Trp Phe Thr Val Ser Trp His Trp His Val Lys Gln Ile Gly Arg

1 5 10 15

Ile Ser Asp Asp Ile Arg Lys Pro Trp Phe Leu Pro Phe Phe Lys Phe

20 25 30

Gly Tyr Lys His Asp Ile Thr Ile Lys Asp Val Tyr Asn Thr Thr Gln

35 40 45

Gly Asp Leu Ser Ala Ala Leu Gly Asp Ala Leu Gln Lys Asn Trp Glu

50 55 60

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

65 70 75 80

Lys Asn Ala Ile Leu Lys Thr Phe Trp Lys Ser Tyr Ser Arg Ser Gly

85 90 95

Ile Ala Leu Gly Phe Gln Phe Val Val Ile Arg Met Leu Gln Pro Ile

100 105 110

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

115 120 125

Ile Gly Trp Trp Leu Ala Thr Gly Val Ile Gly Met Ala Phe Leu Asn

130 135 140

Val Ile Ile Thr His Ser Cys Thr Leu Asp Thr Gln Arg Val Gly Met

145 150 155 160

Arg Val Arg Ile Ala Val Cys Ser Leu Ile Tyr Arg Lys Leu Leu Lys

165 170 175

Leu Ser His Asn Ser Leu Gly Gln Thr Ala Ser Gly Gln Leu Val Asn

180 185 190

Leu Leu Ser Asn Asp Val Gln Arg Phe Asp Leu Ala Ala Gln Phe Ile

195 200 205

His Tyr Ala Trp Ile Met Pro Leu Thr Ala Gly Ile Ser Phe Tyr Ile

210 215 220

Leu Tyr Arg Tyr Val Gly Ile Ile Ala Ala Val Thr Gly Met Val Phe

225 230 235 240

Ile Thr Leu Glu Ser Leu Pro Leu Gln Gly Ser Phe Ser Lys Trp Gln

245 250 255

Gly Lys Leu Arg Tyr Lys Ile Ala Leu Lys Thr Asp Lys Arg Val Lys

260 265 270

Leu Met Ser Glu Ile Thr Ser Gly Ile Gln Val Ile Lys Met Tyr Ala

275 280 285

Trp Glu Lys Pro Phe Glu Lys Val Val Glu Leu Ser Arg Lys Tyr Glu

290 295 300

Ile Asp Ile Ile Thr Thr Thr Ser Tyr Cys Tyr Gly Val Leu Ser Ala

305 310 315 320

Met Gly Ile Phe Thr Glu Arg Met Ile Leu Tyr Ile Thr Val Ile Thr

325 330 335

Phe Val Leu Val Gly Gln Arg Leu Thr Gly Asp Val Val Phe Ser Leu

340 345 350

Ala Gln Leu Phe Asn Thr Val Gln Leu Ile Met Ala Ile Phe Phe Pro

355 360 365

Arg Ala Leu Ser Phe Tyr Ser Glu Ala Lys Val Ser Ile Thr Arg Leu

370 375 380

Glu Asp Phe Leu Leu Leu Asp Glu Asn Glu Lys Ile Pro Glu Pro Glu

385 390 395 400

Lys Ile Ser Asn Pro Asp Glu Leu Gly Glu Ile Gln Leu Thr Asn Ile

405 410 415

Ser Ala Ser Trp Ala Ser Lys Pro Ile Ala Pro Thr Leu Met Asp Leu

420 425 430

Asn Leu His Ile Gln Pro Gly Thr Leu Cys Cys Val Val Gly Asn Val

435 440 445

Gly Cys Gly Lys Ser Ser Leu Leu Gln Leu Leu Leu Arg Glu Leu Pro

450 455 460

Ala Ser Arg Gly Gln Met Lys Ile Asn Gly Lys Ile Ser Tyr Ala Ser

465 470 475 480

Gln Glu Pro Trp Leu Phe Val Ser Asn Val Lys Glu Asn Ile Leu Phe

485 490 495

Gly Lys Pro Phe Leu Lys Asn Arg Tyr His Asp Val Val Lys Val Cys

500 505 510

Ser Leu Glu Arg Asp Phe Lys Gln Phe Pro Phe Gly Asp Arg Ser Leu

515 520 525

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

530 535 540

Asn Leu Ala Arg Ala Val Tyr Thr Glu Ala Asp Ile Tyr Leu Phe Asp

545 550 555 560

Asp Pro Leu Ser Ala Val Asp Thr Lys Val Ala Arg His Leu Phe Asp

565 570 575

Glu Cys Ile Ser Lys Tyr Leu Tyr Gly Lys Thr Arg Ile Leu Val Thr

580 585 590

His Gln Leu Gln Phe Met Lys Lys Ala Asp Leu Ile Ile Ile Ile Asn

595 600 605

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

610 615 620

Asp Leu Asn Ala Leu Gln Gln Glu Pro Glu Ala Asp Asp Lys Glu Lys

625 630 635 640

Glu Lys Val Pro Glu Thr Gly Asp Lys Ile Arg Lys Asn Ser Thr Val

645 650 655

Pro His Phe Gln Ser Met Ser Ser Leu Ala Ser Ser Val Phe Ser Asp

660 665 670

Asp Pro Asn Glu Glu Asp Glu Leu Ile Glu Lys Gly Ala Ile Ser Asn

675 680 685

Ala Thr Tyr Val Glu Tyr Trp Lys Ser Gly Gly Gly Val Leu Phe Leu

690 695 700

Phe Phe Thr Ile Phe Ile Phe Leu Val Ala Gln Met Ile Thr Asn Ala

705 710 715 720

Ser Asp Leu Trp Leu Thr His Trp Thr Asn Asn Glu Ala Lys Arg Tyr

725 730 735

His Leu Ser Asp Ser Leu His Asn Ala Thr Ser Asn Lys Thr Leu Glu

740 745 750

Arg Pro Leu Leu Met Ser Thr Gly Leu Ser Asp Glu Met Ser Thr Ser

755 760 765

Ala Thr Leu Leu Thr Asp Ser Thr Pro Ser Ala Asp Ser Ser Leu Leu

770 775 780

Ser Leu Ser Gln Lys Arg Tyr Leu Thr Asp Leu Ile Arg Asn Ala Thr

785 790 795 800

Leu Glu Glu Leu Asn Asp Ile Pro Ser Thr Glu Tyr Tyr Ile Tyr Val

805 810 815

Tyr Thr Gly Ile Ile Leu Ala Ser Val Val Phe Leu Thr Trp Arg Ser

820 825 830

Phe Leu Tyr Tyr Gln Ile Cys Met Thr Ala Ser Lys Val Leu His Asn

835 840 845

Lys Met Phe Asn Asn Val Leu Gln Ala Pro Met Arg Phe Phe Asp Thr

850 855 860

Asn Pro Ser Gly Arg Ile Leu Asn Arg Phe Ser Lys Asp Met Gly Ala

865 870 875 880

Val Asp Glu Leu Leu Pro Arg Cys Gln Ile Asp Ala Ile Gln Ile Phe

885 890 895

Met Val Met Ile Gly Ile Leu Ala Met Val Phe Ile Val Thr Pro Trp

900 905 910

Met Ile Ile Pro Ala Val Ile Leu Ala Pro Leu Tyr Tyr Phe Phe Arg

915 920 925

Val Val Tyr Leu Thr Ser Ala Gln Ser Leu Lys Arg Leu Glu Gly Val

930 935 940

Ser Arg Ala Pro Val Phe Ser His Ile Ser Ala Ser Leu Tyr Gly Ile

945 950 955 960

Thr Thr Ile Arg Ala Ser Asn Ala Glu Lys Met Ile Thr Thr Glu Phe

965 970 975

Asp Ile Leu Gln Asp Gln His Thr Ser Thr Trp Tyr Leu Phe Ile Val

980 985 990

Ser Ser Thr Ala Phe Gly Phe Tyr Leu Asp Val Met Ser Cys Phe Phe

995 1000 1005

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

1010 1015 1020

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

1025 1030 1035

Leu Thr Gly Met Val Gln Tyr Gly Val Arg Gln Ser Ala Glu Val

1040 1045 1050

Ala Ser Asn Met Ile Ser Val Glu Arg Val Leu Gln Tyr Thr Lys

1055 1060 1065

Leu Asp Lys Glu Gly Pro Phe Glu Thr Leu Pro Gly Lys Lys Pro

1070 1075 1080

Pro Arg Asn Trp Pro Glu Lys Gly Arg Ile Ile Phe Lys Asn Thr

1085 1090 1095

Tyr Leu Lys Tyr Ala Pro Glu Leu Pro Pro Val Leu Lys Asp Leu

1100 1105 1110

Asn Ile Glu Val Asn Ser Gly Glu Lys Ile Gly Ile Val Gly Arg

1115 1120 1125

Thr Gly Ala Gly Lys Ser Thr Leu Ile Ala Ser Leu Phe Arg Leu

1130 1135 1140

Ala Pro Ile Glu Gly Thr Ile Ser Ile Asp Asp Val Asp Thr Ala

1145 1150 1155

Glu Ile Gly Leu Asn Asp Leu Arg Leu Asn Ile Ser Ile Ile Pro

1160 1165 1170

Gln Glu Pro Ile Leu Phe Ser Ala Ser Leu Arg Tyr Asn Leu Asp

1175 1180 1185

Pro Phe Glu Lys His Gly Asp Glu Val Leu Trp Lys Ala Leu Glu

1190 1195 1200

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

1205 1210 1215

Ser Glu Gly Gly Ser Asn Phe Ser Ala Gly Gln Arg Gln Leu Ile

1220 1225 1230

Cys Leu Ala Arg Ala Ile Ile Arg Asn Asn Lys Val Leu Val Met

1235 1240 1245

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

1250 1255 1260

Gln Lys Thr Ile Arg Glu Arg Phe Gln Asp Cys Thr Val Leu Thr

1265 1270 1275

Ile Ala His Arg Leu Asn Thr Ile Met Asp Ser Asp Arg Val Leu

1280 1285 1290

Val Met Asp Ala Gly Gln Ala Met Glu Phe Asp His Pro Tyr Gln

1295 1300 1305

Leu Leu Gln Asn Pro Glu Gly Tyr Phe Ser Lys Met Val Gln Glu

1310 1315 1320

Thr Gly Pro Ala Met Ala Glu Leu Leu Ser Asn Val Ala Lys Asn

1325 1330 1335

Asp Tyr Ile Lys Lys Lys Gly Pro Leu Thr Ser Glu Val Pro Gln

1340 1345 1350

Asn Leu Ala Ile Glu Asp Asn Lys Asp

1355 1360

<210> 46

<211> 1323

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 46

Met Glu Met Thr Lys Glu Lys Tyr Asn Pro Asn Pro Arg Glu Lys Ala

1 5 10 15

Asn Ile Leu Ser Thr Leu Phe Phe Gly Tyr Thr Tyr Asp Val Phe Lys

20 25 30

Lys Gly Leu Ser Lys Thr Leu Glu Val Asp Asp Leu Tyr Asn Pro Leu

35 40 45

Lys Ile Asp Arg Ser Lys Ile Leu Gly Asp His Leu Gln Ser Asn Trp

50 55 60

Asp Lys Gln Leu Glu Lys Ala Lys Lys Thr Asn Ser Lys Pro Ser Leu

65 70 75 80

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

85 90 95

Ile Phe Asp Ile Thr Thr Asp Leu Val Leu Arg Leu Ile Gln Pro Leu

100 105 110

Met Leu Gly Lys Leu Leu Asp His Phe Lys Pro Glu Ala Gln Val Ser

115 120 125

Lys Asn Glu Ala Leu Met Tyr Ala Gly Ile Ile Val Ala Ile Asn Val

130 135 140

Leu Asn Phe Leu Ile Gly Asn Gln Tyr Met Val Glu Gly Phe His Ser

145 150 155 160

Gly Met Arg Ile Arg Ala Ala Cys Cys Ala Val Ile Tyr Arg Lys Ser

165 170 175

Leu Lys Leu Ser Lys Thr Ala Leu Gly Glu Thr Ala Ser Gly Lys Leu

180 185 190

Val Asn Leu Leu Ser Asn Asp Val Ser Arg Phe Asp Leu Val Ser Leu

195 200 205

Leu Ile His Gln Ile Trp Ile Ala Pro Val Leu Ser Leu Ile Val Met

210 215 220

Val Leu Leu Tyr Gln Arg Thr Gly Tyr Ala Gly Ile Val Gly Val Val

225 230 235 240

Ala Val Phe Val Ile Val Pro Ile Gln Thr Tyr Thr Gly Lys Leu Ser

245 250 255

Ala Lys Tyr Arg Lys Gln Thr Ala Met Lys Thr Asp Glu Arg Val Arg

260 265 270

Leu Met Asp Glu Ile Ile Ser Gly Ile Gln Val Ile Lys Met Tyr Ala

275 280 285

Trp Glu Ile Pro Phe Arg Lys Val Ile Arg Ile Ala Arg Arg Asn Glu

290 295 300

Ile Lys Ile Ile Thr Lys Ser Ala Tyr Val Arg Ala Ser Phe Met Ala

305 310 315 320

Leu Asn Leu Phe Thr Thr Arg Leu Ala Leu Phe Cys Thr Leu Leu Thr

325 330 335

Val Ile Leu Ser Asn Glu Pro Ile Thr Ala Ser Met Val Phe Val Val

340 345 350

Met Ser Tyr Phe Asn Val Ile Ser Met Ala Met Ser Thr Met Phe Thr

355 360 365

Arg Gly Val Ser Glu Ile Ala Glu Cys Leu Val Ala Val Arg Arg Ile

370 375 380

Arg Asp Phe Leu Leu Asn Glu Glu Tyr Asp Pro Asn Arg Ser Thr Ser

385 390 395 400

Gly Met Asn Gly Asn Val Lys Ser Val Asp Asp Phe Lys Glu Ile Ile

405 410 415

Ser Leu Lys Asn Leu Thr Val Lys Trp Asn Leu Ser Phe Ser Asp Asn

420 425 430

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

435 440 445

Ile Ile Gly Pro Val Gly Ser Gly Lys Ser Ser Leu Leu Gln Thr Leu

450 455 460

Leu Gly Glu Leu Asp Ile Thr Gln Gly Glu Met Arg Val Gln Gly Gln

465 470 475 480

Ile Ser Tyr Ala Ser Gln Glu Pro Trp Val Phe Ala Ala Thr Val Arg

485 490 495

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

500 505 510

Val Ile Gln Ala Cys Ser Leu Glu Lys Asp Phe Glu Gln Phe Pro Asn

515 520 525

Gly Asp Leu Thr Leu Val Gly Asp Arg Gly Ser Ser Leu Ser Gly Gly

530 535 540

Gln Lys Ala Arg Ile Asn Leu Ala Arg Ala Val Tyr Arg Glu Ala Asp

545 550 555 560

Val Tyr Leu Leu Asp Asp Pro Leu Ser Ala Val Asp Ile His Val Ser

565 570 575

Lys His Leu Tyr Glu Lys Cys Ile Asn Gly Tyr Leu Ala Ser Arg Thr

580 585 590

Arg Ile Leu Val Thr His Gln Val His Tyr Leu Lys Asp Ala Asp Asn

595 600 605

Ile Ile Ile Leu Asn Asn Gly Arg Ile Glu Asp Glu Gly Thr Phe Asn

610 615 620

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

625 630 635 640

Pro Glu Asn Lys Glu Glu Lys Lys Pro Glu Arg Gln Lys Tyr Ser Arg

645 650 655

Gln Leu Ser Gln Arg Ser Arg Lys Asp Ser Met Ser Ser Ile Ile Ser

660 665 670

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

675 680 685

Glu Glu Glu Ala Glu Lys Glu Pro Glu Phe Asp Val Lys Asp Leu Gln

690 695 700

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

705 710 715 720

Met Leu Ala Gly Ser Asn Val Phe Ala Val Phe Ile Cys Val Leu Leu

725 730 735

Tyr Leu Gly Thr Gln Leu Ala Ala Ser Gly Thr Asp Tyr Trp Val Ser

740 745 750

Tyr Trp Val Asn Val Glu Glu Phe Arg Asn Ser Ser Glu Ser Leu Asn

755 760 765

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

770 775 780

Asn Cys Leu Tyr Ile Tyr Ser Val Ile Leu Gly Ala Leu Phe Val Leu

785 790 795 800

Ala Met Thr Arg Ser Phe Phe Phe Tyr Lys Met Ala Met Leu Ser Ser

805 810 815

Lys Lys Leu His Gly Thr Ile Phe Asp Asn Val Ile Asp Ala Thr Met

820 825 830

Arg Phe Phe Asp Thr Asn Pro Gly Gly Arg Ile Leu Asn Arg Phe Ser

835 840 845

Lys Asp Met Gly Ala Val Asp Glu Leu Leu Pro Lys Ala Ile Leu Asp

850 855 860

Ser Ser Gln Ile Leu Leu Ser Met Cys Gly Ser Leu Val Leu Ile Ile

865 870 875 880

Val Val Asn Pro Tyr Phe Leu Ile Leu Ile Gly Val Leu Ser Ala Val

885 890 895

Phe Gly Val Met Arg His Ile Tyr Leu Lys Ser Ser Lys Asn Ile Lys

900 905 910

Arg Leu Glu Gly Ile Met Arg Ser Pro Val Phe Thr His Leu Arg Ala

915 920 925

Thr Ile Glu Gly Leu Thr Thr Ile Arg Ala Phe Gly Ala Gln Ser Thr

930 935 940

Leu Met Asp Glu Phe Asp His His Gln Asp Tyr His Ser Ser Ala Trp

945 950 955 960

Tyr Met Phe Ile Val Ser Ser Thr Ala Phe Gly Phe Tyr Leu Asp Cys

965 970 975

Phe Cys Thr Ile Phe Leu Ala Val Leu Thr Phe Cys Leu Leu Leu Phe

980 985 990

Gly Glu Thr Phe Asn Leu Arg Gly Gly Glu Val Gly Leu Ala Ile Thr

995 1000 1005

Gln Ala Thr Ala Leu Ala Asn Phe Leu Gln Trp Gly Met Arg Gln

1010 1015 1020

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

1025 1030 1035

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

1040 1045 1050

Lys Pro Ala Lys Glu Trp Pro Gln Lys Gly Glu Ile Thr Phe Lys

1055 1060 1065

Asp Thr Cys Leu Arg Tyr Phe Glu Gly Gly Pro Leu Val Leu Lys

1070 1075 1080

His Leu Asn Leu Ser Ile Gln Pro Lys Glu Lys Val Gly Val Val

1085 1090 1095

Gly Arg Thr Gly Ala Gly Lys Ser Ser Leu Ile Gln Ala Leu Phe

1100 1105 1110

Arg Leu Ala Pro Ile Glu Gly Thr Ile Lys Ile Asp Glu Ile Asp

1115 1120 1125

Thr Lys Asp Ile Thr Leu Asn Asp Leu Arg Leu Lys Ile Ser Ile

1130 1135 1140

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

1145 1150 1155

Leu Asp Pro Phe Glu Glu Tyr Thr Asp Glu Val Leu Tyr Lys Ala

1160 1165 1170

Ile Glu Asp Val Glu Leu Arg Asp Pro Ala Asn Val Ile Asn Arg

1175 1180 1185

Leu Glu Asn Arg Val Met Asp Arg Gly Ser Asn Tyr Ser Val Gly

1190 1195 1200

Gln Arg Gln Leu Ile Cys Leu Ala Arg Ala Ile Leu Lys Asn Asn

1205 1210 1215

Lys Val Leu Met Leu Asp Glu Ala Thr Ala Asn Val Asp Pro Gln

1220 1225 1230

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

1235 1240 1245

Cys Thr Val Ile Thr Val Ala His Arg Leu Asn Thr Ile Met Asp

1250 1255 1260

Ser Asp Lys Val Leu Val Met Asp Ala Gly Gln Ile Ser Glu Phe

1265 1270 1275

Asp His Pro His Leu Leu Leu Gln Lys Lys Asp Gly Val Phe Lys

1280 1285 1290

Ser Met Val Cys Glu Thr Gly Arg Ala Thr Ala Glu Gln Leu Lys

1295 1300 1305

Lys Ile Ala Ser Asp Ser Tyr Gln Lys Leu Gln Ala Leu Pro Glu

1310 1315 1320

<210> 47

<211> 1349

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 47

Met Asp Ser Thr Lys Lys His Val Lys Val Ser Pro Glu Glu Ser Ala

1 5 10 15

Ser Pro Phe Ser Val Leu Phe Tyr Trp Trp Ile Leu Pro Phe Leu Lys

20 25 30

Tyr Gly Tyr Gln Asn Asp Ile Gly Met Lys Asp Val Tyr Asn Thr Thr

35 40 45

Gln Ala Asp Gln Ser Gly Pro Leu Gly Asp Glu Leu Gln Glu Asn Trp

50 55 60

Glu Arg Glu Ile Leu Ser Tyr Ile Asp Lys Gln Lys Asn Lys Pro Ser

65 70 75 80

Leu Lys Asn Ala Ile Phe Arg Thr Phe Trp Lys Ser Phe Ile Leu Ser

85 90 95

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

100 105 110

Val Val Leu Ala Lys Tyr Ile Asn Phe Phe Asp Thr Asn Asn Lys Pro

115 120 125

Tyr Leu Gly Trp Ile Trp Gly Cys Gly Val Val Leu Leu Ala Leu Ala

130 135 140

Asn Val Val Leu Tyr His Ser Thr Met Leu Ala Thr Gln Arg Ile Gly

145 150 155 160

Met Arg Ile Arg Thr Ala Val Ser Ser Leu Thr Tyr Arg Lys Leu Leu

165 170 175

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

180 185 190

Asn Leu Met Ser Asn Asp Val Gln Arg Phe Asp Val Cys Ala Ala Ser

195 200 205

Ile His Phe Ile Trp Ile Met Pro Ile Tyr Ala Val Leu Thr Phe Tyr

210 215 220

Ile Leu Tyr Ile Tyr Val Gly Ile Ile Ala Ala Val Thr Gly Met Ala

225 230 235 240

Phe Ile Ser Leu Glu Ser Ile Pro Leu Gln Gly Thr Ile Ser Arg Trp

245 250 255

Gln Gly Val Leu Arg Tyr Lys Ile Ala Leu Arg Thr Asp Lys Arg Ile

260 265 270

Lys Leu Met Ser Glu Leu Thr Ser Gly Ile Gln Val Ile Lys Met Tyr

275 280 285

Ala Trp Glu Lys Pro Phe Glu Lys Ile Val Glu Met Ser Arg Lys Tyr

290 295 300

Glu Ile Asp Val Ile Ala Lys Thr Ser Tyr Leu Tyr Gly Ile Leu Ser

305 310 315 320

Ala Thr Ser Val Phe Thr Glu Arg Leu Ile Leu Tyr Val Thr Leu Ile

325 330 335

Pro Phe Val Leu Leu Gly His Arg Leu Thr Gly Gly Ile Ala Phe Ser

340 345 350

Leu Ala Asn Leu Phe Asn Asn Ile Gln Leu Val Met Ala Ile Asn Phe

355 360 365

Pro Arg Ala Leu Ser Ser Tyr Asn Glu Ala Asn Val Ser Ile Ala Arg

370 375 380

Leu Glu Lys Phe Leu Leu Leu Glu Glu Val Glu Glu Glu Thr Val Val

385 390 395 400

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

405 410 415

Thr Ala Ser Trp Ser Pro Lys Ser Ile Val Pro Thr Leu Ile Asp Ile

420 425 430

Asp Leu His Leu Arg Cys Gly Thr Leu Cys Cys Val Val Gly Asn Val

435 440 445

Gly Ser Gly Lys Ser Ser Leu Leu Gln Leu Phe Leu Arg Glu Leu Pro

450 455 460

Ile Thr Ser Gly Tyr Met Asn Ile Ala Gly Lys Ile Ser Tyr Ala Ser

465 470 475 480

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

485 490 495

Gly Lys Ser Phe Asn Lys Lys Arg Tyr Gln Asp Val Ile Lys Val Cys

500 505 510

Ser Leu Glu Arg Asp Leu Lys Gln Leu Pro Tyr Gly Asp Lys Thr Leu

515 520 525

Val Gly Glu Arg Gly Lys Ser Leu Ser Gly Gly Gln Lys Ala Arg Ile

530 535 540

Asn Leu Ala Arg Ala Val Tyr Thr Glu Ala Asp Ile Tyr Leu Phe Asp

545 550 555 560

Asp Pro Leu Ser Ala Val Asp Thr Lys Val Gly Lys His Leu Phe Asp

565 570 575

Glu Cys Ile Thr Lys Tyr Leu Asn Gly Lys Thr Arg Ile Leu Val Thr

580 585 590

His Gln Leu Gln Tyr Met Lys Lys Ser Asp Leu Ile Val Ile Ile Asn

595 600 605

Asn Gly Lys Ile Asp Lys Val Ala Lys Phe His Asp Leu Thr Glu His

610 615 620

Glu Leu Asn Leu Leu Gln Gln Thr Pro Glu Ile Asp Asp Lys Asp Lys

625 630 635 640

Glu Lys Met Pro Thr Ile Leu Glu Thr Lys Ile Pro Lys Val Ser Ser

645 650 655

Thr Ala Thr Leu Gln Ser Ala Ser Ser Leu Ala Ser Ser Ile Pro Thr

660 665 670

Glu Glu Pro Gly Glu Thr Gly Glu Leu Ile Glu Lys Gly Asp Leu Ser

675 680 685

Thr Ser Leu Tyr Trp Glu Tyr Phe Arg Ser Gly Thr Gly Ile Gly Phe

690 695 700

Leu Leu Phe Thr Gly Phe Met Phe Ile Phe Ser Gln Ile Ile Thr Asn

705 710 715 720

Ala Ser Asp Leu Trp Leu Ser His Trp Thr Asn Val Glu Ala Arg Arg

725 730 735

Tyr Val Ser Thr Leu Asn Leu Ser Thr Glu Phe Ile Ser Thr Thr Ile

740 745 750

Ser Ser Ile Asn Asn His Thr Val Thr Asp Met Leu Lys Ser Val Ala

755 760 765

Phe Thr Asn Thr Asn Pro Gln Glu Ile Ile Thr Thr Ala Asp Pro Ser

770 775 780

Val Ser Ala Ser Lys Leu Ser Ser Asn Met Ser Ser Leu Glu Leu Glu

785 790 795 800

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

805 810 815

Ile Ala Val Val Ile Leu Gln Ile Trp Arg Cys Phe Leu Tyr Tyr Gln

820 825 830

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

835 840 845

Ile Leu Gln Ala Pro Met Arg Phe Phe Asp Thr Asn Pro Ser Gly Arg

850 855 860

Ile Leu Asn Arg Phe Ser Lys Asp Met Gly Ala Val Asp Glu Leu Leu

865 870 875 880

Pro Ser Cys Gln Ile Asp Ala Ile Gln Ile Leu Met Val Phe Val Gly

885 890 895

Ile Leu Val Met Val Val Ile Val Asn Pro Trp Met Ile Leu Thr Thr

900 905 910

Ile Ile Ile Gly Pro Val Leu Phe Ile Leu Arg Lys Met Tyr Leu Lys

915 920 925

Thr Ala Gln Ser Val Lys Arg Leu Glu Gly Ile Ser Lys Ala Pro Val

930 935 940

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

945 950 955 960

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

965 970 975

Gln His Ser Ser Thr Trp Phe Leu Phe Leu Val Ser Gly Arg Val Phe

980 985 990

Gly Phe Tyr Leu Asp Val Ile Cys Cys Ile Phe Leu Ala Ile Val Thr

995 1000 1005

Ile Gln Phe Leu Leu Phe Arg Asp Glu Asn Thr Leu Ser Gly Asn

1010 1015 1020

Val Gly Leu Ala Ile Ser His Ser Tyr Ile Leu Thr Gly Met Val

1025 1030 1035

Gln Met Gly Ile Arg Gln Ser Val Glu Val Ala Ser His Met Ile

1040 1045 1050

Ser Val Glu Arg Ile Leu Gln Tyr Thr Lys Leu Glu Lys Asp Gly

1055 1060 1065

Val Phe Glu Ser Leu Pro Ala Lys Lys Pro Pro Arg Asp Trp Pro

1070 1075 1080

Asn Lys Gly Lys Ile Ile Phe Lys Asn Thr Phe Leu Arg Tyr Ala

1085 1090 1095

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

1100 1105 1110

Ser Gly Glu Lys Val Gly Ile Val Gly Arg Thr Gly Ala Gly Lys

1115 1120 1125

Ser Thr Leu Ile Ala Ser Leu Phe Arg Leu Ala Pro Val Asp Gly

1130 1135 1140

Glu Ile Ile Ile Asp Asp Ile Glu Thr Gly Gly Ile Gly Leu His

1145 1150 1155

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

1160 1165 1170

Phe Ser Ala Ser Val Arg Tyr Asn Leu Asp Pro Phe Glu Lys His

1175 1180 1185

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

1190 1195 1200

Gly Val Val Thr Asp Leu Asn Gln Pro Val Ser Glu Gly Gly Ser

1205 1210 1215

Asn Phe Ser Ala Gly Gln Arg Gln Leu Ile Cys Leu Ala Arg Ala

1220 1225 1230

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

1235 1240 1245

Asn Val Asp Pro Gln Thr Asp Ala Leu Ile Gln Lys Thr Ile Arg

1250 1255 1260

Glu Arg Phe Arg Asp Cys Thr Val Leu Thr Val Ala His Arg Leu

1265 1270 1275

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

1280 1285 1290

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

1295 1300 1305

Asp Gly Tyr Phe Ala Lys Phe Leu Lys Asp Ala Gly Pro Pro Met

1310 1315 1320

Ala Asn Lys Leu Arg Asp Ile Ala Lys Glu Asp Tyr Tyr Lys Lys

1325 1330 1335

His Thr Ser Pro Glu Val Ile Val Asp Asp Asp

1340 1345

<210> 48

<211> 647

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 48

Met Val Lys Val Pro Ala Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser

1 5 10 15

Cys Val Gly Val Trp Gln His Gly Lys Val Glu Ile Ile Ala Asn Asp

20 25 30

Gln Gly Asn Arg Thr Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu

35 40 45

Arg Leu Leu Gly Asp Ala Ala Lys Asn Gln Val Ala Met Asn Pro Ser

50 55 60

Asn Thr Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Lys Phe Asp Asp

65 70 75 80

Pro Lys Ile Gln Gln Asp Ile Gln His Trp Ser Phe Lys Val Ile Asn

85 90 95

His Cys Gly Lys Pro Lys Ile Gln Val Asp Cys Lys Gly Glu Gln Lys

100 105 110

Val Phe Ser Pro Glu Glu Ile Ser Ser Met Val Leu Thr Lys Met Lys

115 120 125

Glu Thr Ala Glu Ala Tyr Leu Gly Thr Thr Val Arg Asp Ala Val Ile

130 135 140

Thr Val Pro Ala Tyr Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp

145 150 155 160

Ala Gly Ala Ile Ala Gly Leu Asn Val Leu Arg Ile Ile Asn Glu Pro

165 170 175

Thr Ala Ala Ala Leu Ala Tyr Gly Leu Asp Lys Asn Leu Lys Gly Glu

180 185 190

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

195 200 205

Ile Leu Thr Ile Asp Glu Gly Ser Leu Phe Glu Val Arg Ser Thr Ala

210 215 220

Gly Asp Thr His Leu Gly Gly Glu Asp Phe Asp Asn Arg Leu Val Asn

225 230 235 240

His Phe Ala Asp Glu Phe Lys Arg Lys Phe Lys Lys Asp Leu Lys Asn

245 250 255

Asn Pro Arg Ala Leu Arg Arg Leu Arg Thr Ala Ala Glu Arg Ala Lys

260 265 270

Arg Thr Leu Ser Ser Ser Ser Glu Ala Thr Ile Glu Ile Asp Ala Leu

275 280 285

Phe Glu Gly Ile Asp Phe Tyr Thr Lys Ile Ser Arg Ala Arg Phe Glu

290 295 300

Glu Leu Cys Ser Asp Leu Phe Arg Gly Thr Leu Gln Pro Val Glu Lys

305 310 315 320

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

325 330 335

Leu Val Gly Gly Ser Thr Arg Ile Pro Lys Ile Gln Gln Leu Leu Gln

340 345 350

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

355 360 365

Ala Val Ala Tyr Gly Ala Ala Val Gln Ala Ala Val Leu Ser Gly Glu

370 375 380

Thr Asp Ser Lys Ile Gln Asp Val Leu Leu Val Asp Val Ala Pro Leu

385 390 395 400

Ser Leu Gly Ile Glu Thr Ala Gly Gly Val Met Ser Asn Ile Ile Ser

405 410 415

Arg Asn Ser Arg Ile Pro Cys Lys Gln Ser Gln Thr Phe Thr Thr Tyr

420 425 430

Ala Asp Asn Gln Pro Ala Val Thr Ile Gln Val Phe Glu Gly Glu Arg

435 440 445

Ala Met Thr Lys Asp Asn Asn Arg Leu Gly Thr Phe Asp Leu Thr Gly

450 455 460

Ile Pro Pro Ala Pro Arg Gly Val Pro Lys Ile Glu Val Thr Phe Asp

465 470 475 480

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

485 490 495

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

500 505 510

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

515 520 525

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

530 535 540

Gly Tyr Val Phe Gln Leu Lys Gln Ala Val Gln Asp Cys Gly Asp Lys

545 550 555 560

Leu Ser Ser Glu Asp Lys Ser Ala Ile Glu Arg Glu Cys Asp Ser Cys

565 570 575

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

580 585 590

Asp Lys Gln Lys Glu Leu Thr Lys Val Cys Ser Pro Ile Met Ser Lys

595 600 605

Leu Tyr Gln Gly Ser Gln Asn Asn Asn Gln His Ser Gly Gly Met Pro

610 615 620

Gly Gly Cys Gly Gln Gln Tyr Gly Gly Phe Gly Gly Arg Gln Ser Gly

625 630 635 640

Pro Thr Val Glu Glu Val Asp

645

<210> 49

<211> 638

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 49

Met Val Leu Pro Thr Asp Leu Thr Ile Met Asp Asp Arg Leu Arg Trp

1 5 10 15

Lys Gln Val Val Gln Lys Lys Val Gln Lys Ala Val Glu Glu Glu Val

20 25 30

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

35 40 45

Asp Lys Lys Phe Phe Ser Gln Leu Ser Gln Leu Phe Gly Ile Ala Cys

50 55 60

Pro Gly Trp Thr Ser Gln Glu Ser Gly Leu Phe Phe Leu Ile Ala Leu

65 70 75 80

Ser Leu Val Ser Arg Ser Met Cys Asp Leu Trp Leu Ile Asn His Gly

85 90 95

Thr Lys Ile Glu Ser Ser Ile Ile Ala Met Asp Pro Thr Leu Phe Lys

100 105 110

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

115 120 125

Asn Asn Val Leu Lys Tyr Ser Ile Gly Ala Leu Lys Ile Gln Leu Arg

130 135 140

Thr Asn Met Thr Arg His Leu Tyr Glu Glu Tyr Leu Lys Asn Tyr Thr

145 150 155 160

Tyr Tyr Arg Ile Ser Asn Leu Asp Asn Arg Ile Ser Asn Ala Asp Gln

165 170 175

Leu Leu Thr Thr Asp Ile Asp Lys Phe Cys Glu Gly Val Thr Asp Leu

180 185 190

Tyr Cys Asn Thr Ala Lys Pro Leu Leu Asp Ile Cys Ile Tyr Val Tyr

195 200 205

Lys Leu Ser Thr Thr Leu Gly Gly Gly Thr Pro Gly Ile Met Leu Val

210 215 220

Tyr Leu Leu Val Ser Gly Val Leu Leu Thr Asn Leu Arg Lys Pro Thr

225 230 235 240

Ala Arg Leu Thr Ala Gly Glu Gln Lys Leu Glu Gly Glu Phe Arg His

245 250 255

Ile Asn Ser Arg Leu Ile Thr His Ser Glu Glu Val Ala Phe Tyr Asn

260 265 270

Gly Asn Ser Arg Glu Lys Ala Thr Leu Met Ala Ser Tyr Asn Lys Leu

275 280 285

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

290 295 300

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

305 310 315 320

Val Val Ser Leu Pro Phe Met Thr Ser Gly His Ser Phe Arg Leu Leu

325 330 335

Gly Thr Asn Glu Arg Ser Arg Leu Tyr Tyr Thr Tyr Gly Arg Met Leu

340 345 350

Val Lys Leu Ala Glu Ala Ile Gly Arg Leu Val Leu Ala Gly Arg Asp

355 360 365

Leu Thr Arg Leu Ala Gly Phe Thr Ala Arg Val Thr Gln Leu Lys Thr

370 375 380

Val Leu Ser Glu Leu Asn Ser Gly Lys Tyr Gln Arg Thr Met Val Ser

385 390 395 400

Gly Ser Glu Ser Leu Gln Ile Asn Gly Gly Lys Leu Ile Phe Arg Asn

405 410 415

Asn Ile Ile Lys Phe His Lys Val Pro Leu Ile Thr Pro Asn Gly Asp

420 425 430

Val Leu Ile Asn Glu Ile Thr Phe Glu Ile Asn Ser Gly Met Asn Val

435 440 445

Leu Val Cys Gly Pro Asn Gly Ala Gly Lys Ser Ser Leu Phe Arg Ile

450 455 460

Leu Gly Glu Leu Trp Pro Leu Phe Gly Gly Glu Leu Thr Lys Pro Pro

465 470 475 480

Arg Gly Lys Leu Phe Tyr Ile Pro Gln Arg Pro Tyr Met Thr Leu Gly

485 490 495

Cys Leu Arg Asp Gln Leu Thr Tyr Pro His Ser Gly Ala Glu Ala Ala

500 505 510

Arg Arg Gly Thr Thr Asp Ala Lys Leu Glu Glu Tyr Leu Gln Arg Val

515 520 525

Gln Leu Gly Tyr Ile Leu Glu Arg Glu Gly Gly Leu Asp Ala Val Ala

530 535 540

Asp Trp Leu Asp Val Leu Ser Gly Gly Glu Lys Gln Arg Ile Ala Met

545 550 555 560

Ala Arg Leu Phe Tyr His Gln Pro Gln Phe Ala Ile Leu Asp Glu Cys

565 570 575

Thr Ser Ala Val Ser Val Asp Val Glu Gly Ser Met Tyr Lys Tyr Cys

580 585 590

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

595 600 605

Trp Gln His His Glu Tyr Val Leu His Leu Asp Gly Arg Gly Gly Tyr

610 615 620

Ser Phe Lys Pro Ile Asp Asn Cys Asp Glu Gln Phe Gly Ser

625 630 635

<210> 50

<211> 754

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 50

Met Pro Thr Val Ile Ser Lys Phe Leu Glu Gln Gly Glu Ser Thr Phe

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Leu Gln Arg Lys Asn Gly Leu Val Lys Asn Lys Lys Leu Lys Gly Arg

65 70 75 80

Leu Lys Asn Ser Ile Pro Asn Phe Asn Leu Ala Phe Ile Leu Gln Phe

85 90 95

Ile Lys Leu Val Arg Ile Met Ile Pro Ser Phe Ile Cys Thr Glu Thr

100 105 110

Val Leu Leu Ser Gly His Thr Thr Phe Leu Phe Leu Arg Thr Phe Leu

115 120 125

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

130 135 140

Met Lys Asp Pro Gln Asn Phe Val Lys Gln Leu Gly Lys Trp Phe Ala

145 150 155 160

Val Ala Ile Pro Ala Thr Phe Ile Asn Ser Met Ile Arg Tyr Leu Glu

165 170 175

Ser Arg Ile Ala Leu Ser Phe Arg Thr Arg Leu Val Asp His Ser Tyr

180 185 190

Lys Leu Tyr Phe Lys Asn Gln Ser Tyr Tyr Arg Val Thr Val Leu Asp

195 200 205

Gly Arg Leu Asp Asn Cys Ala Gln Arg Leu Thr Asp Asp Ile Glu Thr

210 215 220

Val Ala Asn Thr Val Ser His Leu Tyr Gly Gln Ile Thr Lys Pro Cys

225 230 235 240

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

245 250 255

Arg His Ser Asn Leu Val Ile Gly Pro Val Ile Ile Cys Gly Val Val

260 265 270

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

275 280 285

Leu Val Ala Gln Glu Ala Glu Lys Lys Gly Tyr Leu Arg His Val His

290 295 300

Gly Arg Ile Val Ser Asn Ala Glu Glu Ile Ala Phe Tyr Gly Gly His

305 310 315 320

Gln Val Glu Glu Ser Gln Leu Arg Gln Ala Phe Arg Val Leu Ser Lys

325 330 335

His Leu Glu His Met Phe Gly Val Lys Leu Trp Phe Ile Met Leu Glu

340 345 350

Gln Phe Leu Met Lys Tyr Val Trp Ser Gly Ala Gly Ile Ile Val Val

355 360 365

Ser Leu Pro Ile Leu Leu Ala Ala Gly Asn Arg Arg Lys Ile Ser Asn

370 375 380

Lys Ser Leu Leu Ser Ile Pro Asp Phe Ser Ser Lys Ser Glu Ser Leu

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Ala Gly His Thr Ala Arg Val Ala Asn Met Phe Glu Val Leu Glu Glu

450 455 460

Ala Ser Asn Gly Ile Tyr His Lys Thr Leu Val Ala Lys Lys Glu Lys

465 470 475 480

Ser Ala Asp Phe Glu Ile Glu Phe Arg Gly Asp Gln Pro Leu Ala Lys

485 490 495

Gly Lys Ile Ile Tyr Ser Thr Asn Asn Glu Ile Ile Leu Lys Asn Val

500 505 510

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

515 520 525

Glu Leu Thr Pro Gly Gln His Leu Leu Ile Thr Gly Pro Asn Gly Cys

530 535 540

Gly Lys Ser Ser Leu Phe Arg Ile Leu Ser Gly Leu Trp Pro Ile Tyr

545 550 555 560

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

565 570 575

Arg Pro Tyr Met Val Ile Gly Asn Leu Arg Asp Gln Val Ile Tyr Pro

580 585 590

Asp Thr Tyr Thr Asp Met Val Lys Lys Gly Ile Thr Glu Glu Asn Leu

595 600 605

Leu Lys Ile Met Thr Met Val His Leu Asp His Ile Val Glu Arg Asp

610 615 620

Gly Phe Tyr Glu Met Lys Asp Trp Thr Asp Ile Leu Ser Gly Gly Glu

625 630 635 640

Lys Gln Arg Met Ala Ile Ala Arg Leu Phe Tyr His Lys Pro Lys Tyr

645 650 655

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

660 665 670

Phe Ile Tyr Gln Ser Ala Ile Asp Met Gly Ile Thr Leu Leu Thr Ile

675 680 685

Thr His Arg Pro Thr Leu Trp Lys Phe His Thr His Ile Leu Gln Phe

690 695 700

Asp Gly Thr Gly Ser Trp Glu Phe Ser Gln Leu Asn His Thr Ser Arg

705 710 715 720

Leu Thr Leu Lys Lys Glu Lys Glu Asp Leu Leu Lys Ala Glu Asn Asn

725 730 735

Glu Glu Arg Ser Lys Arg Leu Asp Glu Leu Asn Lys Leu Leu Gly Glu

740 745 750

Asp Ser

<210> 51

<211> 921

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 51

Met Ser Lys Lys Arg Gly Ala Lys Lys Gly Lys Asn Leu Asp Asp Asp

1 5 10 15

Phe Glu Glu Thr Ser Ser Ile Ile Ser Glu Lys Glu Lys Ile Asn Ser

20 25 30

Lys Thr Pro Lys Asn Lys Thr Thr Lys Lys Gly Lys Lys Gly Lys Asp

35 40 45

Asp Trp Ser Asp Asp Glu Glu Ile Glu Gln Gly Lys Glu Val Asn Gln

50 55 60

Ser Glu Pro Ala Glu Ser Lys Pro Val Ala Lys Lys Lys Gly Lys Lys

65 70 75 80

Gly Lys Gly Asn Lys Asn Asp Asp Trp Ser Asp Lys Glu Glu Ala Asp

85 90 95

Ile Lys Leu Ser Asp Ser Glu Thr Glu Ser Met Pro Val Ala Val Lys

100 105 110

Lys Ser Ala Lys Lys Asn Lys Lys Lys Lys Asp Asp Trp Ser Asp Lys

115 120 125

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

130 135 140

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

145 150 155 160

Asp Lys Glu Asp Ile Glu Ile Glu Thr Lys Leu Ser Asp Ser Glu Glu

165 170 175

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

180 185 190

Asn Lys Asn Gln Val Arg Asp Glu Ser Pro Asp Ile Glu Glu Met Glu

195 200 205

Ala His Asp Glu Gly Phe Lys Glu Asp Asp Val Ser Glu Glu Glu Ile

210 215 220

Val Lys Pro Val Lys Gln Asp Lys Lys Lys Ser Lys Gly Lys Glu Ser

225 230 235 240

Lys Lys Asp Glu Ser Lys Val Asp Val Glu Val Lys Glu Glu Ile Lys

245 250 255

Glu Ser Lys Glu Asp Ser Pro Ile Val Glu Asp Lys Val Ile Glu Asn

260 265 270

Gly Lys Pro Glu Lys Gln Asn Lys Glu Leu Glu Asn Lys Lys Asp Asn

275 280 285

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

290 295 300

Asp Lys Leu Asp Glu Ser Lys Glu Lys Lys Leu Thr His Lys Glu Lys

305 310 315 320

Lys Lys Met Lys Lys Leu Gln Glu Tyr Glu Lys Gln Met Glu Thr Met

325 330 335

Leu Lys Lys Gly Gly Gln Gly His Ser Glu Leu Asp Ser Asn Phe Thr

340 345 350

Val Ser Gln Thr Gln Lys Thr Ala Gly Gln Leu Ala Ala Phe Glu Asn

355 360 365

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

370 375 380

Asp Leu Phe Val Asn Ala Asn Leu Leu Ile Ala Gln Gly Arg His Tyr

385 390 395 400

Gly Leu Val Gly Pro Asn Gly His Gly Lys Thr Thr Leu Leu Arg His

405 410 415

Val Ala Gln Arg Ala Phe Asp Ile Pro Pro Asn Ile Asp Ile Leu Tyr

420 425 430

Cys Glu Gln Glu Val Val Ala Asp Asp Asn Thr Ala Val Glu Thr Val

435 440 445

Leu Ala Ala Asp Val Lys Arg Asn Asp Leu Leu Ala Glu Cys Lys Lys

450 455 460

Leu Glu Ala Ala Ala Asn Ser Gly Asp Leu Glu Ile Gln Glu Arg Leu

465 470 475 480

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

485 490 495

Pro Arg Ala Arg Arg Ile Leu Ala Gly Leu Gly Phe Asp Lys Glu Met

500 505 510

Gln Asp Arg Ala Thr Lys Asn Phe Ser Gly Gly Trp Arg Met Arg Val

515 520 525

Ser Leu Ala Arg Ala Leu Tyr Ile Glu Pro Thr Leu Leu Leu Leu Asp

530 535 540

Glu Pro Thr Asn His Leu Asp Leu Asn Ala Val Ile Trp Leu Asp Asn

545 550 555 560

Tyr Leu Gln Ala Trp Lys Lys Thr Leu Leu Ile Val Ser His Asp Gln

565 570 575

Ser Phe Leu Asp Asn Val Cys Asn Glu Ile Ile His Leu Asp Asn Lys

580 585 590

Lys Leu Tyr Tyr Tyr Lys Gly Asn Tyr Ser Met Phe Lys Lys Met His

595 600 605

Val Gln Lys Lys Arg Glu Met Ile Lys Glu Tyr Glu Lys Gln Glu Lys

610 615 620

Arg Ile Lys Glu Leu Lys Ser Ser Gly Ser Ser Lys Lys Gln Ala Glu

625 630 635 640

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

645 650 655

Lys Ile Gln Lys Gln Glu Glu Asp Thr Thr Pro Thr Glu Leu Leu Gln

660 665 670

Arg Pro Lys Asp Tyr Leu Val Lys Phe Arg Phe Pro Glu Pro Pro Pro

675 680 685

Leu Gln Pro Pro Val Leu Gly Leu His Asn Thr Arg Phe Ala Tyr Pro

690 695 700

Gly Gln Lys Pro Leu Phe Val Asp Thr Asp Phe Gly Ile Asp Met Ser

705 710 715 720

Ser Arg Val Ala Ile Val Gly Pro Asn Gly Val Gly Lys Ser Thr Phe

725 730 735

Leu Lys Leu Leu Thr Gly Asp Leu Ser Pro Asp Lys Gly Glu Asn Arg

740 745 750

Lys Asn His Arg Leu Arg Ile Gly Arg Phe Asp Gln His Ser Gly Glu

755 760 765

His Leu Thr Ala Glu Glu Thr Pro Ser Glu Tyr Leu Met Arg Leu Phe

770 775 780

Asp Leu Pro Tyr Glu Lys Ala Arg Lys Gln Leu Gly Thr Phe Gly Leu

785 790 795 800

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

805 810 815

Lys Ala Arg Val Ala Leu Ala Glu Leu Cys Leu Asn Ala Pro Asp Val

820 825 830

Leu Ile Leu Asp Glu Pro Thr Asn Asn Leu Asp Ile Glu Ser Ile Asp

835 840 845

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

850 855 860

Ser His Asp Glu Arg Leu Ile Arg Glu Thr Asn Cys Ser Leu Tyr Val

865 870 875 880

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

885 890 895

Arg Lys Glu Leu Leu Glu Ser Leu Gly Glu Val Ile Asn Ser Pro Ser

900 905 910

Ile Ala Ala Asn Ala Ala Val Ala Gln

915 920

<210> 52

<211> 663

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 52

Tyr Gly Leu Leu Gly Ala Ser Gly Cys Gly Lys Thr Thr Leu Leu Ser

1 5 10 15

Ser Ile Val Gly Arg Lys Lys Ile Asp Gly Gly Glu Ile Trp Val Leu

20 25 30

Gly Gly Lys Pro Gly Glu Ala Gly Ser Gly Val Pro Gly Pro Arg Val

35 40 45

Gly Tyr Met Pro Gln Asp Ile Ala Leu Val Gly Glu Phe Thr Val Lys

50 55 60

Asp Ala Ile Tyr Tyr Phe Gly Arg Ile Phe Ser Met Glu Asp Ser Leu

65 70 75 80

Ile Ala Lys Arg Tyr Arg Asn Leu His Thr Leu Leu Glu Leu Pro Pro

85 90 95

Asp Asp Arg Tyr Leu Lys Asn Cys Ser Gly Gly Gln Gln Arg Arg Val

100 105 110

Ser Leu Ala Ala Ser Leu Val His Lys Pro Glu Leu Leu Ile Met Asp

115 120 125

Glu Pro Thr Val Gly Val Asp Pro Val Leu Arg Asp Arg Ile Trp Lys

130 135 140

His Leu Val Asp Ile Thr Lys Lys Asp Asn Thr Ser Val Ile Ile Thr

145 150 155 160

Thr His Tyr Ile Glu Glu Cys Arg Gln Ala Asn Lys Ile Gly Leu Met

165 170 175

Arg Glu Gly Lys Leu Leu Ala Glu Glu Ser Pro Thr Arg Leu Leu Thr

180 185 190

Leu Phe Asn Ser Glu Thr Leu Glu Asp Val Phe Leu Leu Leu Ser Lys

195 200 205

Arg Gln Glu Glu Gly Arg Leu Gln Glu Leu Thr Ser His Arg Val Val

210 215 220

Asp Asp Gln Asn Asn Ser Met Leu Ala Asn Asp Thr Ala Gly Ser Thr

225 230 235 240

Thr Ser Val Ala Thr Ser Val Ser Thr Phe Glu Ile Gly His Gly Ser

245 250 255

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

260 265 270

Asn Lys Ser Arg Met Lys Ala Leu Phe Asp Lys Asn Leu Lys Gln Phe

275 280 285

Tyr Arg Asn Ile Thr Gly Ile Ile Phe Leu Met Thr Phe Pro Ile Leu

290 295 300

Gln Val Gly Val Phe Met Gly Ala Val Gly Gly Asp Ile Arg Ser Ile

305 310 315 320

Pro Leu Gly Ile Val Asn Asp Glu Ala Met Ser Val Thr Cys Pro Gly

325 330 335

Phe Ser Phe Asn Gly Thr Ala Thr Ala Thr Asp Asp Arg Ala Cys Gln

340 345 350

Leu Arg Asn Ile Ser Cys Arg Phe Leu Ser Tyr Leu Asp His Pro Met

355 360 365

Ile Glu Lys Val His Phe Glu Thr Leu Glu Asp Ala Lys Asp Ala Val

370 375 380

Leu His Gly Lys Ile Val Gly Ala Met Tyr Met Ser Ser Asn Phe Thr

385 390 395 400

Ser Phe Leu Glu Glu Arg Ile Asp Lys Gly Lys Asp Ile Glu Lys Asp

405 410 415

Ile Leu Ser Phe Ser Glu Ile Lys Val Trp Met Asp Met Ser Ser Arg

420 425 430

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

435 440 445

Lys Phe Gln Asn Ser Leu Phe Asp Asp Cys Asp Phe Val Pro Gly Phe

450 455 460

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

465 470 475 480

Pro Tyr Thr Val Phe Met Ile Pro Gly Ser Leu Ile Thr Ile Met Phe

485 490 495

Phe Met Gly Ala Ile Met Thr Ser Gln Ile Ile Ile Thr Asp Arg His

500 505 510

Asp Gly Val Trp Asp Arg Ser Ile Val Ala Gly Val Thr Ser Leu Glu

515 520 525

Ile Thr Ile Thr His Leu Val Leu Gln Ala Ser Ile Cys Ile Ile Gln

530 535 540

Thr Ala Glu Leu Leu Val Val Val Tyr Leu Ile Tyr Gln Gln Glu Tyr

545 550 555 560

Ser Gly Ser Leu Trp Leu Met Tyr Val Met Val Tyr Leu Gln Gly Ile

565 570 575

Cys Gly Met Ala Tyr Gly Phe Trp Val Ser Val Ile Ser Thr Asp His

580 585 590

Ser Met Ala Asn Thr Val Leu Thr Gly Ile Phe Leu Pro Met Met Met

595 600 605

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

610 615 620

Trp Gly Thr Asn Arg Cys Arg Lys Phe Leu Pro Gly Val Glu Lys Ala

625 630 635 640

Ala Tyr Ala Phe Ser Glu Gln Pro Leu Thr Arg Gly Glu Ile Val Asp

645 650 655

Lys Ser Ala Gly Cys Thr Leu

660

<210> 53

<211> 827

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 53

Met Val Leu Pro Thr Ile Ala Asn Leu Glu Gln Asn Pro Glu Ala Leu

1 5 10 15

Pro Phe Thr Glu Asp Phe Asn Lys Leu Phe Glu Ala Phe Phe Gln Thr

20 25 30

Tyr Ser Arg Gln Lys Glu Leu Glu Thr Leu Ala Glu Asp Met Thr Ser

35 40 45

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

50 55 60

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

65 70 75 80

Ala Trp Lys Leu Ala Asp Ala Ala His Ala Arg Glu Gln Leu Ala His

85 90 95

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

100 105 110

Ile Glu Phe Lys Asn Lys Met Asn Gln Asp Thr Asp Glu Tyr Gly Leu

115 120 125

Leu Gly Ala Ser Gly Cys Gly Lys Thr Thr Leu Leu Asn Cys Ile Val

130 135 140

Gly Arg Lys Arg Leu Asn Ser Gly Glu Leu Trp Val Leu Gly Gly Thr

145 150 155 160

Pro Gly Ser Arg Gly Ser Gly Val Pro Gly Pro Arg Val Gly Tyr Met

165 170 175

Pro Gln Glu Thr Ala Leu Asn Gly Glu Phe Thr Ile Arg Glu Thr Leu

180 185 190

Lys Tyr Phe Gly Trp Ile Ser Arg Met Thr Thr Asp Glu Val Glu Ala

195 200 205

Lys Val Asp Phe Phe Ile Asn Phe Leu Met Leu Pro Asp Ala Asp Arg

210 215 220

Gln Val Lys Thr Leu Ser Gly Gly Gln Gln Arg Arg Val Ser Leu Ala

225 230 235 240

Ala Thr Leu Leu His Glu Pro Glu Leu Leu Ile Leu Asp Glu Pro Thr

245 250 255

Val Gly Val Asp Pro Leu Leu Arg Thr Asn Ile Trp Asn His Leu Val

260 265 270

Glu Ile Thr Lys Tyr Gly Arg Thr Thr Val Ile Ile Thr Thr His Tyr

275 280 285

Ile Asp Glu Thr Arg Gln Ala His Leu Ile Gly Leu Met Arg Gly Gly

290 295 300

Tyr Phe Leu Ala Glu Glu Ser Pro Glu Arg Leu Ile Thr Gln Phe Gly

305 310 315 320

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

325 330 335

Met Gly Lys Arg Arg Arg Ser Ser Ile Ala Lys Ser Val Val Glu Thr

340 345 350

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

355 360 365

Asp Asp Asp Leu Gly Glu Ile Ser Gly Glu Phe Gly Asp Ser Val Ser

370 375 380

Val Thr Ser Arg Thr Gly Arg Arg Val Ser Ile Ala Pro Glu Pro Thr

385 390 395 400

Ala Glu His Ile Ile Pro Glu Leu Pro Pro Asp Glu Glu Pro Glu Val

405 410 415

Ser Phe Lys Asp Tyr Phe Lys Phe Val Gln Ser His His Met Arg Ala

420 425 430

Leu Ile Trp Lys Asn Phe Leu Trp Met Trp Arg Asn Ile Pro Met Met

435 440 445

Met Phe Ile Ile Gly Leu Pro Ile Ser Gln Thr Ile Leu Phe Cys Trp

450 455 460

Thr Ile Gly His Asp Pro Gln Gly Ile Lys Val Ser Val Val Asn Asn

465 470 475 480

Glu Ile Asn Phe Pro Asp Glu Thr Cys His Gln Gln Pro Leu Asn Cys

485 490 495

Asn Gly Thr Lys Ile Ser Cys Asn Tyr Leu Asn Glu Val Ala Lys Ser

500 505 510

Tyr Ser Ile Thr Trp Glu Phe Met Lys Thr Glu Glu Glu Ala Arg His

515 520 525

Arg Val Glu Arg Gly Lys Ser Trp Gly Val Val Val Val Pro His Asn

530 535 540

Phe Thr Asp Ala Leu Trp Ser Arg Ile Glu Asn Ser Arg Asp Thr Pro

545 550 555 560

Pro Glu Asp Ile Met Ala Ser Thr Ile Ser Val Tyr Glu Asp Lys Ser

565 570 575

Asn Glu Asn Ile Ala Thr Phe Leu Thr Arg Asp Met Leu Tyr Gly Phe

580 585 590

Leu Thr Phe Ile Ser Glu Phe Val Lys Ser Cys Gly Tyr Asn Glu Lys

595 600 605

Ser Val Gly Val Pro Ile Arg Tyr Asn Glu Pro Ile Tyr Gly Tyr Gln

610 615 620

Asp Pro Asp Phe Thr Asp Phe Ala Ala Pro Gly Val Val Leu Thr Ile

625 630 635 640

Ile Phe Phe Met Ala Val Ala Leu Thr Ser Gly Ser Met Ile Met Glu

645 650 655

Arg Asn Glu Gly Ile Leu Glu Arg Ser Leu Val Asn Gly Ile Ser Gly

660 665 670

Thr Glu Leu Leu Phe Ser Gln Ile Ile Thr Gln Phe Val Val Met Leu

675 680 685

Gly Gln Ser Ile Ala Val Leu Leu Val Ala Phe Ile Leu Phe Gly Leu

690 695 700

Thr Gln Arg Gly Asp Trp Thr Thr Val Thr Thr Leu Thr Val Leu Ala

705 710 715 720

Gly Ile Cys Gly Met Cys Phe Gly Phe Val Val Ala Cys Ser Cys Glu

725 730 735

Asn Glu Arg Ser Ala Thr Tyr Leu Ala Met Gly Ser Phe Leu Pro Ile

740 745 750

Val Met Leu Cys Gly Ile Ile Trp Pro Ile Glu Ala Met His Lys Tyr

755 760 765

Ile Ser Trp Met Ser Val Val Leu Pro Leu Thr Gln Ala Thr Glu Ser

770 775 780

Leu Arg Cys Ile Leu Ala Arg Gly Trp Pro Ile Ser Asn Pro Thr Val

785 790 795 800

Tyr Asn Gly Phe Ile Ser Leu Thr Ile Trp Ile Phe Ile Phe Leu Thr

805 810 815

Ile Ser Ile Leu Leu Leu Lys Phe Lys Lys Gly

820 825

<210> 54

<211> 613

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 54

Met Gly Glu Asn Thr Val Cys Asp Ile Leu Gly Thr Val Asp Pro Pro

1 5 10 15

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

20 25 30

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

35 40 45

Gly Phe Arg His Ile Ser Tyr Ser Val Tyr Asp Gly Ile Leu Phe Ser

50 55 60

Arg Thr Lys Arg Lys Leu Leu Asp Asp Ile Thr Gly Asn Phe Pro Gly

65 70 75 80

Gly Gln Leu Thr Ala Ile Met Gly Pro Ser Gly Ala Gly Lys Thr Cys

85 90 95

Leu Met Asn Ile Leu Ser Gly Tyr Ala Thr Thr Gly Ile Leu Gly Glu

100 105 110

Val Leu Val Asn Asp Glu Pro Arg Asn Asn Val Ala Phe Arg Lys Gln

115 120 125

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

130 135 140

Leu Glu Ser Met Arg Val Ala Ala Asp Leu Met Leu Thr Ala Ser Lys

145 150 155 160

Thr Asp Lys Asp Lys Lys Ile Ala Ser Ile Leu Lys Ser Val Thr Leu

165 170 175

Trp Glu Ile Lys His Thr Arg Thr Asp Ala Leu Ser Gly Gly Gln Lys

180 185 190

Lys Arg Leu Ser Val Ala Leu Glu Leu Leu Arg Asp Pro Gln Val Met

195 200 205

Phe Phe Asp Glu Pro Thr Ser Gly Leu Asp Ser Leu Asn Ser Ile Arg

210 215 220

Leu Val Lys Leu Leu Lys Glu Met Ser Glu Ser Gly Lys Thr Ile Ile

225 230 235 240

Cys Thr Ile His Gln Pro Val Pro Arg Tyr Leu Asn Cys Ser Ile Thr

245 250 255

Leu Leu Glu Val Ala Ser Gly Asp Tyr Gly Asp Phe Thr His Ala Leu

260 265 270

Ser Thr Lys Ser Asn Asn Gly Ile Asn Glu Ser Tyr Ala Lys Leu Ser

275 280 285

Ala Ser Thr Met Lys Leu Asn Ser Ile Lys Glu Pro Asn Ile Gly Glu

290 295 300

Asp Phe Leu Ala Ala Thr Lys Arg Thr Ile Val Asn Glu Ile Glu Tyr

305 310 315 320

Thr Trp Asp Glu Cys Gly Leu Asn Ser Tyr Pro Thr Ser Thr Ile Asn

325 330 335

Gln Phe Val Val Leu Thr Lys Arg Ser Phe Leu Met Leu Ser Arg Asp

340 345 350

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

355 360 365

Phe Leu Gly Ile Leu Tyr Phe Gly Val Gly Leu Asp Ala Ala Asn Ile

370 375 380

Asn Asp Asn Phe Ser Phe Met Phe Phe Thr Val Met Phe Leu Met Met

385 390 395 400

Thr Ala Phe Asn Cys Val Val Thr Thr Phe Pro Ser Glu Leu Pro Ile

405 410 415

Ile Ile Lys Glu His Phe Asn Lys Trp Tyr Ala Ile Arg Ser Tyr Tyr

420 425 430

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

435 440 445

Leu Tyr Ala Thr Ile Thr Tyr Leu Cys Thr Gln Gln Pro Met Glu Trp

450 455 460

Ser Arg Val Ser Ser Phe Leu Phe Ile Cys Ile Leu Val Ser Val Ile

465 470 475 480

Ser Gln Ser Trp Ala Leu Val Val Gly Ser Cys Leu Ser Val Thr Asn

485 490 495

Gly Val Val Ile Gly Pro Phe Phe Leu Leu Pro Phe Ile Met Phe Ser

500 505 510

Gly Tyr Phe Val Gln Leu Arg Asp Cys Pro His Gln Leu Lys Trp Met

515 520 525

Phe Asp Ile Ser Phe Pro Arg Tyr Ala Leu Glu Gly Leu Val Leu Thr

530 535 540

Ile Phe Gly Tyr Asp Arg Gly Lys Leu Pro Cys Glu Ser Lys Asp Phe

545 550 555 560

Cys Leu Tyr Val Tyr Pro Glu Gln Phe Ile Lys Asp Lys Asp Met Glu

565 570 575

Asn Ala Ser Tyr Thr Ile Gly Val Met Cys Leu Met Gly Leu Ile Val

580 585 590

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

595 600 605

Asp Arg Arg Arg Arg

610

<210> 55

<211> 625

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 55

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

1 5 10 15

Ala Val Ile Leu Lys Asp Val Ser Met Leu Val His Ser Gly Glu Val

20 25 30

Leu Ala Val Leu Gly Ser Lys Gly Ser Gly Lys Lys Ala Leu Leu Asp

35 40 45

Val Ile Ser Arg Arg Ala Gln Gly Pro Ile Arg Gly Gln Ile Tyr Leu

50 55 60

Asn Asn His Pro Val Ser Met Cys Leu Phe Gln Gln Lys Cys Ala Tyr

65 70 75 80

Val Thr His Lys Cys Asp Phe Ile Pro Gly Leu Asn Val Glu Gln Thr

85 90 95

Leu Tyr Tyr Thr Pro Thr Lys Phe Thr Gly Tyr Leu Lys Met Ser Lys

100 105 110

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

115 120 125

Cys Val Glu Asp Leu Thr Lys Ser Glu Tyr Arg Arg Leu Met Ile Gly

130 135 140

Val Gln Leu Ile Lys Asn Pro Val Val Leu Leu Leu Asp Glu Pro Thr

145 150 155 160

Trp Asp Leu Asp Pro Leu Asn Thr Tyr Leu Ile Ile Ser Ile Leu Ser

165 170 175

Asn Ala Ala Lys Lys Tyr Gly Thr Ala Ile Ile Leu Thr Met Glu Lys

180 185 190

Pro Arg Ser Asp Val Phe Pro Phe Leu Asp Arg Val Leu Tyr Leu Cys

195 200 205

Leu Gly Asp Val Val Tyr Thr Gly Gly Thr Lys Gln Met Leu Glu Tyr

210 215 220

Phe Asn Val Ile Gly Phe Pro Cys Pro Gln Leu Glu Asn Pro Leu Met

225 230 235 240

Tyr Tyr Leu Cys Leu Ser Thr Val Asp Arg Arg Ser Arg Glu Arg Phe

245 250 255

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

260 265 270

Glu Gly Val Ile Phe Gln Lys Ser Pro Ser Met Ile Ser Pro Asn Pro

275 280 285

Asn His Glu His Gly Gln His Glu Lys Val Pro Phe Met His Gly Arg

290 295 300

Pro Gly Arg Phe Ser Thr Gly Trp Thr Ile Tyr Val Arg Leu Leu Ala

305 310 315 320

Ala Thr Val Ser Phe Lys Arg Ala Gly Leu Arg Gln Thr Phe Leu Arg

325 330 335

Thr Phe Ala Leu Pro Leu Tyr Phe Phe Leu Met Trp Leu Phe Tyr Arg

340 345 350

Glu Met Lys Asp Trp Gln His Thr Phe Ile Ser Arg Asn Gly Leu Ile

355 360 365

Leu Asn Cys Leu Cys Cys Val Tyr Phe Val Gly Ile Ile Asn Thr Ile

370 375 380

Leu Ile Tyr Pro Ile Tyr Arg Thr Arg Tyr Tyr Gln Asp Thr Gln Glu

385 390 395 400

Gly Leu Tyr Gly Gly Thr Leu Phe Leu Leu Thr Tyr Asn Leu Val Ser

405 410 415

Leu Pro Phe Ser Phe Leu Ser Thr Val Leu Ser Ala Val Ile Ile Phe

420 425 430

Arg Lys Leu Ile Ile Ser Phe Asp Asn Pro Val Asp Phe Val Tyr Phe

435 440 445

Thr Leu Ile Leu Trp Ala Cys Tyr Ile Phe Ala Glu Gln Gln Thr Met

450 455 460

Ala Ile Leu Met Ile Val Lys Asp Tyr Met Lys Ala Ala Ile Phe Ser

465 470 475 480

Ile Tyr Leu Thr Val Val Cys Ile Thr Leu Gly Ser Gly Ile Leu Arg

485 490 495

Ser Met Lys Gly Leu Pro Glu Trp Leu Tyr Tyr Ala Thr Tyr Gly Thr

500 505 510

Gln Ala Arg Tyr Ala Ala Ala Tyr Leu Asn Arg Lys Val Phe Thr His

515 520 525

Ser Ala Leu Ser Lys Ala Leu Pro Phe Asp Leu Leu His Asn Cys Thr

530 535 540

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

545 550 555 560

Cys Arg Tyr Ala Ser Gly Gln Ala Phe Leu Thr Glu Arg Tyr Ser Arg

565 570 575

Asp Pro Thr Glu Val Ile Phe Ser Gly Val Leu Glu Glu Asp Phe Asn

580 585 590

Val Gly Leu Thr Cys Gly Phe Ala Leu Gly Met Ile Val Phe Asn Leu

595 600 605

Phe Leu Tyr Leu Ile Pro Leu Pro Ser Phe Val Lys Ala Lys Phe Arg

610 615 620

Glu

625

<210> 56

<211> 940

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 56

Met Met Arg Ser Arg Thr Pro Ser Gln Gly Gly Gly Ser Gln Gly Phe

1 5 10 15

Glu Met Glu Arg Lys Tyr Ser Val Pro Ser Asn Pro Glu Ser Arg Ala

20 25 30

Phe Ser Gly Gly Thr Thr Ser Glu Asp Leu His Ala Trp Ser Ile Tyr

35 40 45

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

50 55 60

Asp Lys Ser Pro Leu Pro Tyr Gly Asn Phe Gln Leu Arg Asp Thr Thr

65 70 75 80

Val Gln Ser Ile Leu Ser His Pro Arg Tyr Gly Pro Lys Ser Ala Leu

85 90 95

Gly Ser Asn Met Tyr Thr Tyr Leu Lys Phe Gly Leu Pro Arg Val Phe

100 105 110

Pro Pro Asn His Asn Gly Ser Asn Arg Ser Gly Thr Pro Gln His Phe

115 120 125

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

130 135 140

Gly Ser His Gly Pro Arg Asp Gly Ser Ser Gly Tyr Asp Ser Ser Asp

145 150 155 160

Asn Glu Thr Ser His Asn Tyr Lys Gln Asn Arg Lys Tyr Arg Ser Asp

165 170 175

Pro Asp Phe Arg Met Gln Asn Val Tyr His Ala Glu Gln Thr Ser Pro

180 185 190

Gly Ile Pro Leu Ala Ala Met His Gln Gly Asp Ile Arg His Ser Asn

195 200 205

Ser Gln Trp Asn Arg Asn Lys Ser Ile Ser Glu Ala Asn Leu Leu Ala

210 215 220

Leu Gly Tyr Asn Arg Ser Cys His Asn Ser Glu Arg His Leu Ile Asp

225 230 235 240

Pro Arg Arg Asn Ser Val Ala Asp Tyr Gly His His Asn His Asp Val

245 250 255

Val Asp His Ser Val Met Gly His Met Gly Arg Pro Met Ser Lys Thr

260 265 270

Glu Ser His Phe Ser Val Pro His Ser Arg Arg Gly Pro Pro Ser Ile

275 280 285

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

290 295 300

Met Phe Pro His Leu Gln Ala His Gly Leu Gly Ile Phe Pro Ser Ser

305 310 315 320

Gln Ser Cys Thr Lys Ser Arg Gln His Leu Leu Leu Asn Glu Ile Ser

325 330 335

Phe Glu Ile Arg Gly Gly Glu Ile Met Ala Ile Met Thr Thr Ser Glu

340 345 350

Glu Glu Gly Thr Ala Leu Leu Asp Ile Ile Ala Gly Phe Ser Ser Pro

355 360 365

Ala Leu Gly Thr Ile Phe Leu Asn Gly His Ser Val Arg Ala His Thr

370 375 380

Leu Lys Ser Arg Val Ala Tyr Val Gln Asn Asp Leu Asn Leu Cys Lys

385 390 395 400

Asp Met Thr Val Val Gln Thr Leu Arg Leu His Tyr Asp Leu Lys Lys

405 410 415

Pro Thr Glu Lys Leu Gly Tyr Leu Lys Ile Glu Ser Met Asp Arg Ile

420 425 430

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

435 440 445

Val Ser Met Met Thr Ile Ser Glu Arg Arg Arg Leu Asn Val Ala Cys

450 455 460

His Leu Ile Leu Asp Thr Asp Ile Val Leu Leu Asp Gln Pro Thr Lys

465 470 475 480

Ser Met Asp Ile Phe Asp Thr Phe Phe Leu Val Glu Tyr Leu Lys Gln

485 490 495

Trp Ala Ser Gly Gly Ala Gly Ser Thr Leu Gly Arg Ile Val Ile Leu

500 505 510

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

515 520 525

Leu Leu Val Ser Ala Gly Arg Thr Met Tyr Ser Gly Arg Arg Arg Asp

530 535 540

Met Leu Pro Tyr Phe Ala Leu Val Glu Tyr Pro Cys Pro Ala Phe Lys

545 550 555 560

Asn Pro Ser Asp Tyr Tyr Leu Asp Leu Val Thr Leu Asp Asp Leu Ala

565 570 575

Ala Glu Ala Met Leu Glu Ser Ser Gln Arg Ile Glu Gln Leu Ala Glu

580 585 590

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

595 600 605

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

610 615 620

Ala Leu Phe Thr Lys Ser Met Ile Tyr Thr Gln Pro Ala Thr Phe Leu

625 630 635 640

Ser Trp Leu Thr Val Ile Val Leu Ser Ala Ser Leu Ser Leu Ile Leu

645 650 655

Gly Ala Ile Phe Trp Asp Ile Pro Ser Thr Asp Pro Gln Leu Ile Leu

660 665 670

Asn Asp Arg Tyr Gly Tyr His Tyr Ser Val Met Cys Ile Val His Trp

675 680 685

Pro Leu Leu Leu Ala Met Thr Val Asn Glu Val Arg Arg Asn Arg Lys

690 695 700

Val Ile Glu Arg Asp Ile Lys Asp Gly Leu Tyr Gly Arg Val Thr Tyr

705 710 715 720

Ile Ile Thr Lys Ser Ile Ile Asn Ile Phe Pro Ser Leu Phe Val Trp

725 730 735

Leu Ile Tyr Val Val Pro Ser Tyr Ser Met Thr Gly Leu Tyr Met Gln

740 745 750

His Leu Asn Asn Tyr Asp Gly Phe Tyr Ile Tyr Ile Gly Val Met Leu

755 760 765

Leu Tyr Leu Ser Cys Ile Gln Ile Phe Leu Met Ala Phe Ile Tyr Thr

770 775 780

Val Pro Leu Ser Asn Thr Ala Thr Ile Phe Cys Gly Thr Val Leu Ser

785 790 795 800

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

805 810 815

Met Tyr Leu Gln Trp Ile Glu Lys Ile Ser Pro Ser Glu Trp Leu Ile

820 825 830

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

835 840 845

Gln Gly Thr Ile Thr Thr Leu Cys Arg Asn Lys Gln Ile Gln His Gln

850 855 860

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

865 870 875 880

Leu Lys Ser Phe Gly Tyr Leu Lys Ser Asp Asn Leu Thr Phe Asp Tyr

885 890 895

Gly Asn Pro Val Ile Ala Met Gly Val Phe Tyr Cys Ile Phe Phe Val

900 905 910

Ile Ser Cys Phe Met Phe Ala Leu Asn Leu Cys Arg Ser Arg Arg Arg

915 920 925

Arg Arg Gln Asp Thr Lys Asn Asp Ala Asn Lys Pro

930 935 940

<210> 57

<211> 678

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 57

Met Leu Val Leu Ser Val Ser Gly Ser Gln Thr Asn Ile Tyr Asn Gly

1 5 10 15

Ser Ser Pro Ile Thr Ile Ser Gly Ala Gly Ala Leu Arg Lys Val Pro

20 25 30

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

35 40 45

Leu Pro Lys Arg Pro Pro Val Asp Ile Thr Phe Thr Asp Leu Ser Tyr

50 55 60

Ser Val Ser Glu Gly Arg Lys Arg Gly Tyr Lys Thr Ile Leu Lys Cys

65 70 75 80

Ile Ser Gly Lys Cys Lys Ser Gly Glu Leu Thr Ala Ile Met Gly Pro

85 90 95

Ser Gly Ala Gly Lys Ser Thr Ile Met Asn Ile Leu Ala Gly Tyr Lys

100 105 110

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

115 120 125

Leu Arg Arg Phe Arg Lys Met Ser Cys Tyr Ile Met Gln Asp Asp Cys

130 135 140

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

145 150 155 160

Leu Lys Leu Gly Lys Thr Val Thr Leu Ser Glu Lys Lys Val Val Ile

165 170 175

Asn Glu Ile Ile Glu Asn Leu Gly Leu Gln Thr Cys Ile Asp Ile Asn

180 185 190

Ser Ser Asn Leu Ser Gly Gly Gln Arg Lys Arg Leu Ser Ile Gly Leu

195 200 205

Glu Leu Val Asn Asn Pro Pro Val Met Phe Phe Asp Glu Pro Thr Ser

210 215 220

Gly Leu Asp Ser Ser Ser Cys Phe Gln Cys Leu Cys Leu Leu Lys Ser

225 230 235 240

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

245 250 255

Ala Arg Leu Phe Glu Met Phe Asp His Leu Tyr Met Met Ala Glu Gly

260 265 270

Gln Cys Ile Tyr Arg Gly Pro Val Leu Gly Leu Val Pro Phe Leu Ser

275 280 285

Ser Met Gly Leu Asn Cys Pro Ser Tyr His Asn Pro Ala Asp Tyr Val

290 295 300

Met Glu Val Ala Cys Gly Glu His Gly Asp Tyr Val Gln Lys Leu Val

305 310 315 320

Val Ala Val Asn Ala Gly Arg Cys Thr Lys Phe Ala Thr Pro Asp His

325 330 335

Arg Ser Ser Lys Ile Val Ser Asn Asp Ile Ala Lys Glu Ala Asn Gly

340 345 350

Lys Asn Ser Ser Gly Asp Val Ile Ser Val Pro Asn Gly Ser Val Lys

355 360 365

Pro Thr Thr Pro Thr Thr Pro Val Thr Cys Thr Thr Ser Leu Leu Asp

370 375 380

Ser Ser Glu Asn Leu Ser Pro Thr Glu Lys Asn Gly Phe Ser Thr Thr

385 390 395 400

Gly Leu Gln Gln Phe Thr Ile Leu Leu Lys Arg Ser Met Tyr Met Ile

405 410 415

Leu Met Asp Lys Thr Leu Thr Arg Met Arg Leu Val Ser His Phe Val

420 425 430

Ile Gly Cys Leu Ile Gly Leu Ile Tyr Tyr Asp Ile Gly Gln Asp Ala

435 440 445

Ala Lys Val Thr Ser Asn Ala Gly Cys Leu Phe Phe Cys Val Met Phe

450 455 460

Met Met Tyr Thr Ala Met Met Pro Thr Ile Leu Thr Phe Pro Leu Glu

465 470 475 480

Met Ser Val Thr Val Arg Glu His Leu Asn Tyr Trp Tyr Ser Leu Lys

485 490 495

Ala Tyr Tyr Met Ala Lys Thr Leu Ala Asp Ile Pro Phe Gln Val Val

500 505 510

Met Thr Leu Cys Tyr Ile Ile Gly Val Tyr Phe Ile Thr Ser Gln Pro

515 520 525

Leu Asp Val Thr Arg Phe Gly Met Ile Leu Leu Val Thr Val Leu Thr

530 535 540

Ala Leu Val Ser Gln Ser Phe Gly Leu Leu Ile Gly Ala Ala Phe Asn

545 550 555 560

Ile Glu Gly Gly Val Phe Leu Gly Pro Ile Ser Thr Ile Pro Met Val

565 570 575

Leu Phe Ser Gly Phe Phe Thr Asn Leu Asn Asp Ile Pro Phe Tyr Leu

580 585 590

Arg Trp Leu Pro Tyr Leu Ser Tyr Leu Lys Tyr Gly Phe Glu Ala Cys

595 600 605

Met Ile Ala Ile Tyr Gly Leu Asp Arg Pro Lys Leu Thr Cys Asn Ile

610 615 620

Glu Tyr Cys His Phe Lys Tyr Pro Lys Lys Phe Leu Glu Gln Met Ser

625 630 635 640

Met Lys Asp Asp Met Val Ser Tyr Phe Ile Asp Val Gly Val Leu Gly

645 650 655

Gly Leu Phe Ile Phe Leu Arg Val Ile Ala Tyr Phe Met Leu Arg Ile

660 665 670

Lys Leu Met Gln Asn Arg

675

<210> 58

<211> 549

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 58

Met Gly Pro Ser Gly Ala Gly Lys Thr Ser Leu Leu Asn Ile Leu Thr

1 5 10 15

Gly Tyr Gln Val Thr Gly Thr Thr Gly Thr Ile Lys Cys Asn Ser Ser

20 25 30

Ser Arg Lys Gln Lys Gly Val Leu Gln Tyr Lys Lys Glu Ser Cys Tyr

35 40 45

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

50 55 60

Met Met Ile Ala Ser Lys Leu Lys Ile Ala Asn Met Ala Lys Lys Ala

65 70 75 80

Arg Glu Phe Leu Ile Asn Glu Ile Leu Thr Asn Leu Ser Leu Leu Lys

85 90 95

Ala Lys Asn Thr Arg Cys Gln Ser Leu Ser Gly Gly Gln Lys Lys Arg

100 105 110

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

115 120 125

Asp Glu Pro Thr Thr Gly Leu Asp Ser Ala Ser Thr Thr Gln Cys Val

130 135 140

Asp Leu Leu Lys Lys Leu Ala Asn Gly Gly Arg Ile Val Ile Cys Thr

145 150 155 160

Ile His Gln Pro Asn Thr Gln Thr Tyr Glu Met Phe Asp Gln Val Tyr

165 170 175

Met Leu Ala Lys Gly Arg Cys Val Tyr Gln Gly Pro Ser Thr Asn Thr

180 185 190

Val Pro Phe Leu Ala Ser Val Gly Leu His Cys Pro Gln Tyr His Asn

195 200 205

Pro Ala Asp Tyr Ile Met Glu Val Val Ser Gly Glu Tyr Gly Asp His

210 215 220

Ile Asp Gln Leu Ala Val Ala Ala Gln Asp Lys Lys Trp Gln Asn Ile

225 230 235 240

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

245 250 255

Asn Ile Ile Tyr Ser Asp Glu Asn Val Thr Leu Ser Lys Ser Pro Ser

260 265 270

Glu Trp Lys Arg Phe Phe Ile Leu Leu Gln Arg Ser Ser Val Gln Leu

275 280 285

Tyr Arg Asp Trp Thr Ile Ser Gln Leu Lys Leu Val Leu His Leu Leu

290 295 300

Val Gly Leu Phe Leu Gly Ile Thr Phe Gln Asn Cys Gly Arg Asp Ala

305 310 315 320

Thr Lys Val Ile Ser Asn Leu Gly Phe Leu Gln Val Gly Ile Val Tyr

325 330 335

Leu Ala Tyr Thr Ser Met Met Pro Ala Val Leu Lys Phe Pro Thr Glu

340 345 350

Leu Val Ile Leu Lys Lys Glu Ser Phe Asn Asn Trp Tyr Lys Leu Thr

355 360 365

Thr Tyr Tyr Ala Ala Phe Leu Val Phe Asp Ile Pro Gln Gln Met Leu

370 375 380

Phe Ser Thr Val Tyr Cys Ile Gly Cys Tyr Phe Val Ser Asp Gln Pro

385 390 395 400

Leu Glu Val Asp Arg Phe Phe Ser Val Leu Phe Val Leu Val Leu Ala

405 410 415

Ser Leu Ser Ser Ser Gly Phe Gly Leu Ile Leu Gly Thr Ile Thr Asn

420 425 430

Pro Ile Asn Gly Val Phe Phe Gly Ala Val Gly Leu Cys Phe Phe Ile

435 440 445

Ser Val Gly Gly Phe Phe Ile Met Phe Thr His Met Ser Asn Val Met

450 455 460

Tyr Leu Phe Ser Tyr Ile Ser Tyr Ile Ser Phe Ser Val Glu Gly Val

465 470 475 480

Met Gln Ala Ile Tyr Gly Tyr Gly Arg Gly Gln Leu His Cys Pro Glu

485 490 495

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

500 505 510

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

515 520 525

Cys Thr Phe Leu Thr Phe Arg Thr Ile Ala Phe Val Thr Leu Lys Arg

530 535 540

Lys Leu Ala Asn Pro

545

<210> 59

<211> 604

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 59

Ser Lys Leu Ile Leu Arg Ser Val Asn Gly Ser Phe Gln Ala Gly Gln

1 5 10 15

Leu Thr Ala Ile Met Gly Pro Ser Gly Ala Gly Lys Ser Thr Leu Leu

20 25 30

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

35 40 45

Ile Asn Gly Ala Thr Arg Asn Leu Lys Gln Phe Arg Lys Met Ser Arg

50 55 60

Tyr Ile Met Gln Glu Asp Met Ile Gln Pro Leu Leu Thr Val Glu Glu

65 70 75 80

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

85 90 95

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

100 105 110

Asp Lys Ala Lys Arg Thr Gly Thr Ser Arg Leu Ser Gly Gly Glu Arg

115 120 125

Lys Arg Leu Ser Ile Ala Leu Glu Leu Leu Asn Asn Pro Pro Val Leu

130 135 140

Phe Leu Asp Glu Pro Thr Thr Gly Leu Asp Asp Leu Ser Cys Ser Gln

145 150 155 160

Cys Ile Ser Leu Leu Lys Lys Ile Ala Glu Gly Gly Arg Thr Val Ile

165 170 175

Cys Ser Ile His Thr Pro Ser Ala Lys Ile Phe Ser Gln Phe Asp Asn

180 185 190

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

195 200 205

Glu Val Val Ser Tyr Leu Ser Lys Val Gly Ile Glu Cys Pro Lys Thr

210 215 220

Tyr Asn Pro Ala Asp Phe Ile Ile Glu Val Cys Cys Asn Glu Tyr Gly

225 230 235 240

Asn Phe Gln Glu Arg Met Val Ser Val Ile Asp Asn Gly Arg Asn Ile

245 250 255

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

260 265 270

Asn Asp Ser Leu Glu Asn Arg Ser Ser Ile Ser Ser Ile Ser Ile Arg

275 280 285

Glu Glu Val Tyr Ala Asp Leu Ser Ser Ser Asp Phe Asn His Glu Ser

290 295 300

Ser Trp Phe Ser Gln Phe Trp Ile Leu Thr Thr Arg Leu Trp Met Gln

305 310 315 320

Met Trp Arg Asp Lys Thr Tyr Leu Ile Met Arg Thr Val Leu Tyr Ile

325 330 335

Ile Leu Ala Leu Leu Ile Gly Ser Leu Tyr Tyr Lys Met Gly Gln Asp

340 345 350

Gly Ser Lys Thr Ile Phe Asn Phe Gly Phe Tyr Tyr Cys Cys Ile Ile

355 360 365

Phe Phe Met Tyr Ile Pro Met Met Pro Ile Leu Leu Gln Phe Pro Gln

370 375 380

Glu Ile Gln Leu Val Lys Arg Glu His Phe Asn Lys Trp Tyr Arg Leu

385 390 395 400

Ser Ala Tyr Phe Ser Ala Leu Ser Phe Ser Thr Val Pro Val Gln Leu

405 410 415

Cys Leu Gly Val Val Tyr Val Ser Cys Val Tyr Leu Leu Thr Asp Gln

420 425 430

Pro Leu Glu Phe Arg Arg Met Ser Met Phe Phe Phe Ile Cys Ile Leu

435 440 445

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

450 455 460

Lys Val Val Asn Ala Val Phe Met Gly Pro Val Ser Ala Val Pro Phe

465 470 475 480

Met Leu Leu Ala Val Tyr Gly Phe Gly Ser Gly Tyr Asp Thr Ile Pro

485 490 495

Ser Ile Ile Lys Phe Phe Met His Phe Ser Tyr Leu Arg Tyr Ser Leu

500 505 510

Glu Gly Leu Ile His Ala Met Leu Lys Asp Arg Glu Lys Leu Ser Cys

515 520 525

Pro Asp Thr Glu Glu Tyr Cys Ile Tyr Thr Asp Leu Asn Leu Phe Val

530 535 540

Arg Asp Met Gly Met Glu Asn Thr Ile Tyr Trp Val Asp Val Leu Val

545 550 555 560

Leu Ile Phe Ile Leu Ile Leu Phe Arg Gly Gly Ser Tyr Tyr Leu Leu

565 570 575

Arg Gln Arg Leu Thr Pro Asn Lys Thr Phe Arg Ala Leu Gln Tyr Ile

580 585 590

Gly Arg Leu Val Lys Ser Gln Phe Gly Leu Ala Arg

595 600

<210> 60

<211> 617

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 60

Glu Lys Lys Ile Leu Lys Asn Val Ser Gly Met Leu Arg Ser Gly Glu

1 5 10 15

Leu Cys Ala Ile Met Gly Pro Ser Gly Ala Gly Lys Ser Thr Leu Leu

20 25 30

Asn Ile Leu Thr Gly Tyr Lys Thr Ile Gly Val Lys Gly His Val Leu

35 40 45

Met Asn Gly Ser Asp Arg Asp Leu Ser Gln Phe Arg Lys Leu Ser Ala

50 55 60

Tyr Ile Met Gln Asp Asn Gln Leu His Ala Asn Leu Arg Val Asp Glu

65 70 75 80

Ala Met Ala Val Ala Ala Ala Leu Lys Ile Gly Ala Lys Ser Pro Lys

85 90 95

Asp Arg Glu Asp Ile Ile Asn Glu Ile Leu Asp Thr Leu Gly Leu Leu

100 105 110

Asp His Lys Lys Thr Met Thr Ser Gly Leu Ser Gly Gly Gln Lys Lys

115 120 125

Arg Leu Ser Ile Ala Leu Glu Leu Val Ser Asn Pro Pro Val Met Phe

130 135 140

Phe Asp Glu Pro Thr Ser Gly Leu Asp Ser Ser Ser Cys Phe Gln Cys

145 150 155 160

Ile Ser Leu Leu Lys Thr Leu Ala Lys Gly Gly Arg Thr Ile Ile Cys

165 170 175

Thr Ile His Gln Pro Ser Ala Arg Leu Phe Glu Met Phe Asp Gln Leu

180 185 190

Tyr Thr Leu Ala Asp Gly Gln Cys Val Tyr Gln Gly Ser Thr Thr Phe

195 200 205

Leu Val Pro Phe Leu Ala Ser Leu Gln Leu Gln Cys Pro Ser Tyr His

210 215 220

Asn Pro Ala Ser Tyr Ile Ile Glu Val Ala Cys Gly Glu Tyr Gly Asp

225 230 235 240

His Thr Arg Thr Leu Val Asn Ala Ile Glu Asn Gly Lys Asn Asp Ile

245 250 255

Arg Asp Ala Ser Gln Ile Val Gly Phe Lys Met Ser Asp Gly Leu Asn

260 265 270

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

275 280 285

Glu Leu Ser Lys Glu Gly Ser Thr Val Asn Gly Asn Arg Lys Ser Asn

290 295 300

Asn Val Asn Glu Asp Thr Gly Ile Asp Ser Lys Asp Val Glu Lys Ala

305 310 315 320

Asn Val Asp Ser Ala Leu Leu Asn Thr Ser Val Val Val Lys Gln Pro

325 330 335

Arg Tyr Gly Asn Ser Glu Phe Gln Gln Phe Phe Ile Ile Leu Lys Arg

340 345 350

Ala Leu Leu Phe Ser Arg Arg Asp Trp Thr Leu Met Tyr Leu Arg Leu

355 360 365

Phe Ala His Ile Leu Val Gly Phe Leu Ile Gly Ala Leu Tyr Phe Lys

370 375 380

Ile Gly Asn Asp Gly Ser Lys Val Leu Ser Asn Leu Gly Phe Leu Phe

385 390 395 400

Phe Asn Met Leu Phe Leu Met Tyr Thr Ser Met Thr Ile Thr Ile Leu

405 410 415

Ser Phe Pro Leu Glu Met Pro Val Leu Leu Lys Glu His Phe Asn Arg

420 425 430

Trp Tyr Ser Leu Arg Ser Tyr Tyr Leu Ala Ile Thr Ile Ser Asp Met

435 440 445

Pro Phe Gln Thr Ile Phe Cys Ile Leu Tyr Val Thr Ile Val Tyr Phe

450 455 460

Met Thr Ser Gln Pro Leu Asp Pro Ala Arg Phe Gly Met Phe Leu Leu

465 470 475 480

Ser Ser Leu Leu Val Ser Phe Val Ala Gln Ser Val Gly Leu Val Val

485 490 495

Gly Ala Ala Met Asn Val Gln Asn Gly Val Phe Leu Ala Pro Val Met

500 505 510

Ser Val Pro Phe Leu Leu Phe Ser Gly Phe Phe Val Ser Phe Asp Ala

515 520 525

Ile Pro Ile Tyr Leu Arg Trp Ile Thr Tyr Leu Ser Tyr Ile Arg Tyr

530 535 540

Gly Phe Glu Gly Thr Ala Leu Ala Thr Tyr Gly Tyr Asn Arg Pro Asn

545 550 555 560

Leu Asp Cys Phe Ala Gln Tyr Cys His Phe Lys Lys Ala Leu Thr Thr

565 570 575

Leu Glu Glu Leu Asp Met Asp Lys Ser Ser Tyr Trp Phe Asp Ile Leu

580 585 590

Ala Leu Val Val Ile Phe Phe Phe Leu Arg Val Ser Ala Tyr Leu Phe

595 600 605

Leu Lys Trp Lys Leu Lys Ser Asn His

610 615

<210> 61

<211> 570

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 61

Thr Lys Glu Ile Leu His Gln Val Gly Gly Arg Phe Gln Pro Gly Gln

1 5 10 15

Leu Ile Ala Ile Met Gly Pro Ser Gly Ala Gly Lys Ser Thr Leu Leu

20 25 30

Asp Val Leu Ser Gly Tyr Arg Ile Arg Gly Val Thr Gly Ser Val Tyr

35 40 45

Val Asn Gly Gln Pro Arg Ser Leu Lys Glu Phe Arg Lys Thr Ser Cys

50 55 60

Tyr Ile Thr Gln Asp Asp Arg Leu Gln Pro Leu Leu Thr Val Asp Glu

65 70 75 80

Asn Met Trp Val Ala Ala Asp Leu Lys Leu Pro Ser Ser Val Ser Asn

85 90 95

Ser Asn Lys Arg Ser Ile Ile Asn Asn Ile Leu Lys Thr Leu Asn Leu

100 105 110

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

115 120 125

Lys Arg Leu Ser Ile Ala Leu Glu Leu Val Asn Asn Pro Met Val Met

130 135 140

Phe Leu Asp Glu Pro Thr Thr Gly Leu Asp Ser Ser Ser Cys Ser Thr

145 150 155 160

Cys Ile Thr Leu Leu Lys Lys Leu Thr Met Gln Gly Lys Thr Ile Ile

165 170 175

Cys Thr Ile His Gln Pro Ser Ala Thr Leu Phe Ala Met Phe Asp Gln

180 185 190

Val Tyr Val Val Gly Asn Gly Tyr Cys Leu Tyr Gln Gly Ser Thr Asn

195 200 205

Lys Leu Val Pro Phe Leu Gln Asp Cys Gly Phe Pro Cys Pro Gln Tyr

210 215 220

His Asn Pro Ala Asp Tyr Val Ile Glu Leu Ala Cys Ile Glu His Gly

225 230 235 240

Pro Glu Lys Pro Glu Tyr Met Arg Glu Arg Ile Gln Asn Gly Gln Ser

245 250 255

Tyr Gln Tyr Phe Asn Glu Pro Glu Lys Met Lys Ser Thr Ile Cys Asn

260 265 270

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

275 280 285

Ser Phe Pro Thr Ser Gln Phe His Gln Leu Gly Val Leu Met Arg Arg

290 295 300

Gly Phe Ile Lys Ala Tyr Arg Asp Lys Met Leu Thr Tyr Leu Arg Ile

305 310 315 320

Gly Thr Asn Ile Ile Val Gly Leu Met Leu Gly Ser Leu Tyr Trp Lys

325 330 335

Ala Gly Ser Asp Gly Ser Lys Val Leu Asp Asn Phe Asn Leu Leu Phe

340 345 350

Ala Ile Leu Leu His His Met Met Ser Thr Met Met Leu Thr Ile Leu

355 360 365

Thr Phe Pro Gln Glu Met Ser Ile Leu Ile Lys Glu His Phe Asn Arg

370 375 380

Trp Tyr Ser Leu Lys Met Tyr Tyr Thr Ser Val Thr Ile Val Asp Ile

385 390 395 400

Pro Val Ser Val Ile Gly Cys Phe Leu Phe Ser Leu Ile Ile Tyr Tyr

405 410 415

Met Thr Asp Gln Pro Asn Asp Lys Ala Arg Phe Leu Ile Phe Phe Val

420 425 430

Thr Cys Met Leu Val Val Leu Val Ala Gln Gly Val Gly Leu Thr Ile

435 440 445

Gly Ala Tyr Phe Asp Val Val Asn Gly Thr Phe Val Gly Pro Thr Leu

450 455 460

Met Val Pro Met Met Met Phe Ser Gly Phe Gly Val Arg Leu Arg Asp

465 470 475 480

Leu Pro Thr Ile Met Tyr Tyr Gly Ser Phe Thr Ser Tyr Leu Arg Tyr

485 490 495

Gly Leu Glu Gly Val Val Gln Ala Val Tyr Gly Met Asp Arg Gly Ile

500 505 510

Leu Gln Cys Pro Glu Asp Lys Phe Cys Phe Tyr Lys Tyr Pro Lys Thr

515 520 525

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

530 535 540

Ile Ala Leu Leu Leu Phe Leu Phe Val Leu Arg Ile Ala Ala Tyr Val

545 550 555 560

Val Leu Arg Tyr Lys Leu Ala Ser Val Arg

565 570

<210> 62

<211> 507

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 62

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

1 5 10 15

Lys Pro Leu Thr Gln Leu Asn Thr Ile Ala Lys Lys Pro Pro Ile Asp

20 25 30

Val Glu Phe Cys Asp Leu Thr Tyr Ser Ile Ser Asp Ser Ser Tyr Lys

35 40 45

Gly Gly Trp Arg Gln Leu Leu Lys Ser Ile Asn Gly Lys Phe Arg Ser

50 55 60

Gly Glu Leu Thr Ala Ile Leu Gly Pro Ser Gly Ala Gly Lys Ser Thr

65 70 75 80

Leu Leu Asn Ile Leu Ala Gly Tyr Met Thr Ala Gly Val Lys Gly Ser

85 90 95

Val Lys Ile Asn Gly Lys Pro Arg Asp Met Arg Ile Phe Thr Lys Leu

100 105 110

Ser Ser Tyr Ile Met Gln Glu Asp Leu Val Gln Pro Arg Leu Ser Val

115 120 125

Arg Glu Ser Met Met Val Ala Ala Asn Leu Lys Leu Ser Ala Ser Ile

130 135 140

Gly His Thr Gln Lys Val Ala Val Val His Glu Val Ile Gln Leu Leu

145 150 155 160

Gly Leu Glu Lys Cys Tyr Asp Thr Lys Thr Glu Tyr Leu Ser Gly Gly

165 170 175

Gln Arg Lys Arg Leu Ser Val Ala Leu Glu Leu Val Asn Asn Pro Pro

180 185 190

Val Ile Phe Leu Asp Glu Pro Thr Thr Gly Leu Asp Asn Val Ser Ile

195 200 205

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

210 215 220

Val Ile Cys Thr Ile His Gln Pro Pro Ala Ser Leu Phe Gln Ile Phe

225 230 235 240

Asp Gln Val Tyr Ile Met Ala Asn Gly Tyr Cys Val Tyr Asn Gly Ser

245 250 255

Pro Asn Gln Leu Val Pro Phe Met Ser Ser Val Asn Cys Val Cys Pro

260 265 270

Glu Thr Ser Thr Pro Ala Asp Phe Ile Ile Glu Val Ile Gln Thr Asn

275 280 285

Gln Asp Asn Ile Pro Ile Leu Gln Asn Gln Ile Gln Asn Gly Lys Ile

290 295 300

Asn Met Lys Asp Lys Lys Leu Lys Pro Leu Gln Ser His Lys Thr Leu

305 310 315 320

Gly Ile Tyr Glu Ile Tyr Gln Glu Thr Thr Gln Thr Gly Met His Ile

325 330 335

His Asp Ile Glu Tyr Pro Thr Ser Phe Trp Thr Gln Phe Thr Val Leu

340 345 350

Leu Cys Arg Met Ala Leu Gln Met Lys Arg Asn Lys Ser Met Trp Ile

355 360 365

Ile Gln Phe Phe His His Val Ile Ser Ala Thr Leu Val Gly Gly Ile

370 375 380

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

385 390 395 400

Tyr Cys Val Thr Ile Asn Val Phe Phe Val Tyr Thr His Val Met Val

405 410 415

Pro Val Leu Leu Cys Ile Ser Ile Glu Val Lys Leu Leu Lys Arg Glu

420 425 430

Tyr Phe Asn Arg Trp Phe Ser Leu Lys Pro Tyr Phe Leu Ala Ser Thr

435 440 445

Ile Val Asn Ile Pro Met Leu Val Gly Tyr Gly Met Ile Phe Ile Ser

450 455 460

Ile Val Phe Phe Met Thr Gly Gln Pro Ile Glu Trp Gln Arg Phe Leu

465 470 475 480

Met Phe Thr Met Ile Ala Ile Asn Val Gly Phe Cys Ser Gln Gly Leu

485 490 495

Gly Tyr Ala Ile Gly Ser Asn Cys Gly Ile Leu

500 505

<210> 63

<211> 709

<212> PRT

<213> 玉米根萤叶甲（Diabrotica virgifera）

<400> 63

Met Thr Pro Val Gly Pro Gly Ala Lys Thr Lys Ile Glu Leu Glu Arg

1 5 10 15

Ser His Ser Leu Leu Pro Ile Arg His Ile Pro Asn Gly Asp Leu Ala

20 25 30

Val Ala Ala Val Val Ser Leu Gly Gly Gln Arg Ala Thr Val Ser Leu

35 40 45

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

50 55 60

Glu Arg Glu Thr Phe Leu Arg Gln Gly Glu Arg Gln Ile Leu Ser Glu

65 70 75 80

Asn Glu Ile Ile Leu Asp Arg Trp Ser Thr Phe His His Arg Val Ala

85 90 95

Leu Val Arg Val Pro Phe Asn Pro Val Pro Val Val Val Asp Thr Cys

100 105 110

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

115 120 125

Ala Val Leu Gly Phe Leu Ile Arg Val Pro Asn Glu Phe Asp Ile Tyr

130 135 140

Ala Ser Ala Ser Leu Ala Lys Lys Arg Gln Arg Thr Tyr Ser Arg Trp

145 150 155 160

Ser Pro Ile Glu Glu Gly Val Thr Leu Val Trp Glu Asn Leu Ser Val

165 170 175

Tyr Ser Ser Thr Thr Lys Asn Gly Gln Met Gln His Lys Gln Ile Ile

180 185 190

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

195 200 205

Gly Ser Ser Gly Ala Gly Lys Ser Thr Leu Met Thr Ala Leu Gly Tyr

210 215 220

Arg Thr Glu Gly Ser Ile Leu Thr Glu Gly Asn Ile Leu Ile Asn Gly

225 230 235 240

Arg Gln Ile Gly Asp Tyr Met Lys Tyr Leu Ser Gly Phe Met His Gln

245 250 255

Glu Asp Met Leu Leu Ser Tyr Leu Thr Val Arg Glu His Met Asn Ile

260 265 270

Met Ala Asn Leu Lys Leu Asp Arg Arg Leu Ser Gly Asn Asp Lys Lys

275 280 285

Gln Leu Ile Tyr Asp Ile Leu Arg Gln Leu Gly Leu Met Lys Cys Ile

290 295 300

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

305 310 315 320

Glu Lys Lys Arg Leu Ala Phe Ala Thr Glu Leu Leu Thr Asp Pro Leu

325 330 335

Leu Leu Phe Cys Asp Glu Pro Thr Thr Gly Leu Asp Ser Tyr Ser Ala

340 345 350

Gln Lys Leu Val Val Ile Met Asn Gln Met Ala Ile Thr Gly Lys Thr

355 360 365

Ile Leu Cys Thr Ile His Gln Pro Ser Ser Asp Ile Phe Ala Met Phe

370 375 380

Ser Gln Leu Ile Leu Val Ala Asp Gly Arg Ile Ala Tyr Met Gly Ser

385 390 395 400

Thr Asn Asn Ala Leu Asp Phe Phe Glu Arg Met Gly Tyr Val Cys Pro

405 410 415

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

420 425 430

Pro Gly Tyr Glu Asp Asn Cys Arg Gln Thr Val Lys Arg Ile Cys Asp

435 440 445

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

450 455 460

Tyr Glu Leu His Met Gly Arg Val Ala Thr Gln Arg Lys Phe Glu Leu

465 470 475 480

Arg Gln Asn Phe Lys Glu Ile Arg Trp Leu Ser Lys Leu Phe Trp Leu

485 490 495

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

500 505 510

Met Lys Ile Ala Gln Arg Met Leu Ile Gly Phe Ile Val Gly Phe Cys

515 520 525

Tyr Leu Gly Thr Asp Ala Leu Thr Gln Asn Gly Val Gln Ser Val Thr

530 535 540

Gly Ile Ile Phe Met Phe Val Ser Glu Asn Thr Phe Asn Pro Met Tyr

545 550 555 560

Ser Val Leu His Gln Phe Pro Ser Tyr Leu Pro Leu Phe Leu Arg Glu

565 570 575

Tyr Lys Ser Gly Leu Tyr His Pro Ala Thr Tyr Tyr Leu Ser Arg Ile

580 585 590

Leu Ser Leu Leu Pro Gly Phe Ile Leu Glu Pro Leu Ile Phe Val Ile

595 600 605

Thr Ala Tyr Trp Leu Ala Gly Leu Arg Thr Ser Gly Tyr Ala Phe Leu

610 615 620

Met Thr Leu Ser Val Ile Val Leu Thr Met Asn Val Ser Ser Ala Cys

625 630 635 640

Gly Val Met Phe Ser Asn Ala Phe Asp Ser Val Pro Thr Ala Leu Ala

645 650 655

Tyr Leu Val Pro Phe Asp Tyr Ala His Met Ile Thr Ser Gly Val Phe

660 665 670

Val Lys Leu Ser Thr Leu Pro Leu Val Phe Ser Trp Ile Lys Tyr Leu

675 680 685

Ser Trp Leu Met Tyr Ser Thr Glu Ala Leu Tyr Ile Leu Gln Trp Lys

690 695 700

Asp Val Thr Asn Ile

705

Claims (14)

1.一种用于检测作物田间对Cry3类杀昆虫蛋白质具有抗性的昆虫的方法，所述方法包括：

a.提供来自昆虫的样品；

b.测定所述样品中存在的核酸；以及

c.检测与SEQ ID NO:1相比改变的DvABCB1序列或降低的DvABCB1基因的转录物水平，其中所述改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。

2.如权利要求1所述的方法，其进一步包括在检测到Cry3抗性昆虫时，改变作物田间的有害生物综合管理策略。

3.如权利要求1所述的方法，其进一步包括在检测到Cry3抗性昆虫时，向作物田间喷洒杀昆虫剂。

4.一种用于检测作物田间对Cry3类杀昆虫蛋白质具有抗性的昆虫的方法，所述方法包括：

a.提供来自昆虫的样品；

b.测定所述样品中存在的核酸；以及

c.检测与SEQ ID NO:1具有小于95％序列同一性的改变的DvABCB1序列或降低的DvABCB1基因的转录物水平，其中所述改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。

5.如权利要求4所述的方法，其进一步包括在检测到Cry3抗性昆虫时，改变作物田间的有害生物综合管理策略。

6.如权利要求4所述的方法，其进一步包括在检测到Cry3抗性昆虫时，向作物田间喷洒杀昆虫剂。

7.一种用于检测作物田间对Cry3类杀昆虫蛋白质具有抗性的昆虫的方法，所述方法包括：

a.提供来自昆虫的样品；

b.测定所述样品中存在的核酸；以及

c.检测与SEQ ID NO:1相比具有缺失的改变的DvABCB1序列或降低的DvABCB1基因的转录物水平，其中所述改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。

8.如权利要求7所述的方法，其进一步包括在检测到Cry3抗性昆虫时，改变作物田间的有害生物综合管理策略。

9.如权利要求7所述的方法，其进一步包括在检测到Cry3抗性昆虫时，向作物田间喷洒杀昆虫剂。

10.一种用于检测作物田间对Cry3类杀昆虫蛋白质具有抗性的昆虫的方法，所述方法包括：

a.提供来自昆虫的样品；

b.测定所述样品中存在的多肽；以及

c.检测与SEQ ID NO:1的翻译序列相比改变的DvABCB1氨基酸序列或降低的DvABCB1基因的转录物水平，其中所述改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。

11.如权利要求10所述的方法，其进一步包括在检测到Cry3抗性昆虫时，改变作物田间的有害生物综合管理策略。

12.如权利要求10所述的方法，其进一步包括在检测到Cry3抗性昆虫时，向作物田间喷洒杀昆虫剂。

13.一种用于筛选昆虫群体的试剂盒，其中所述试剂盒包含用于检测与SEQ ID NO:1相比改变的DvABCB1序列或降低的DvABCB1基因的转录物水平的装置，其中所述改变的DvABCB1序列或降低的DvABCB1的转录物水平显示Cry3抗性昆虫。

14.一种DNA构建体，其包含编码来自玉米根萤叶甲(Diabrotica virgifera)的ABC类受体的多核苷酸，其中所述多核苷酸可操作地连接到异源调节元件，并且其中所述ABC类受体与SEQ ID NO:40-63中的任一个具有至少95％序列同一性。

Images