δ－内毒素基因及其使用方法

专利名称：δ－内毒素基因及其使用方法
技术领域：
本发明涉及分子生物学领域。所提供的是编码杀虫蛋白质的新基因。这些蛋白质及编码它们的核酸序列可用于制备杀虫制剂以及生产转基因的抗害虫植物。
背景技术：
苏云金芽孢杆菌是形成孢子的革兰氏阳性土壤细菌，特征在于它能产生对某些目和种的昆虫有特异性毒性、但对植物和其它非靶向生物体无害的晶状内含物。因此，含苏云金芽孢杆菌菌株或它们的杀虫蛋白质的组合物可用作环境可接受的杀虫剂，以抑制农业有害昆虫或针对多种人或动物疾病的昆虫载体。
来自苏云金芽孢杆菌的晶体(Cry)蛋白质(δ-内毒素)具有主要针对鳞翅类、双翅类和鞘翅类幼虫的有效杀虫活性。这些蛋白质还显示出抗膜翅目、同翅目、虱、食毛目和螨等有害目以及诸如线虫、扁形动物(Platyhelminthes)和Sarcomastigorphora等其它无脊椎动物目的活性(Feitelson(1993)苏云金芽孢杆菌家族树，高级基因工程杀虫剂中(The Bacillus Thuringiensis family tree.InAdvanced Engineered Pesticides).Marcel Dekker，Inc.，New York，N.Y.)。这些蛋白质主要基于其杀虫活性而最初被划分为CryI至CryV。主要的种类是鳞翅类特异的(I)、鳞翅类和双翅类特异的(II)、鞘翅类特异的(III)、双翅类特异的(IV)以及线虫类特异的(V)和(VI)。所述蛋白质进一步被划分为亚家族，各个家族内更密切相关的蛋白质被指派上分类字母，诸如Cry1A、Cry1B、Cry1C等。在各亚类内更紧密相关的蛋白质则命名为诸如Cry1C1、Cry1C2等。
最近出现了一种针对Cry基因的新命名法，其基于的是氨基酸序列同源性而非昆虫靶特异性(Crickmore等(1998)Microbiol.Mol.Biol.Rev.62807-813)。在该新的分类法中，每种毒素被分配了一个独特的名称，其中合并了第一等级(阿拉伯数字)、第二等级(大写字母)、第三等级(小写字母)和第四等级(另一阿拉伯数字)。在新的分类中，罗马数字被转换成第一等级的阿拉伯数字。序列同一性小于45％的蛋白质具有不同的第一等级，而对于第二和第三等级而言这一标准分别为78％和95％。
晶体蛋白质在被昆虫中肠摄取和溶解前不呈现杀虫活性。被摄取的毒素原在昆虫消化道内被蛋白酶水解成活性的有毒分子。(H_fte和Whiteley(1989)Microbiol.Rev.53242-255)。此毒素与靶目标幼虫中肠内的顶端刷状边缘受体结合并插入顶端膜中，产生离子通道或孔，导致幼虫死亡。
δ-内毒素通常具有5个保守的序列结构域和三个保守的结构性结构域(参阅，例如，de Maagd等(2001)Trends Genetics17193-199)。第一个保守的结构性结构域由7个α螺旋组成，并涉及膜插入和孔形成。结构域II由排列成希腊语钥匙构形的三个β折叠片组成，而结构域III由“果冻-卷”形式的两个反向平行β折叠组成(de Maagd等(2001)同上文)。结构域II和III涉及受体识别和结合，并因此被认为是毒素特异性的决定区。
因为昆虫可造成的破坏，所以一直需要发现新形式的苏云金芽孢杆菌δ-内毒素。
发明简述本发明提供了赋予细菌、植物、植物细胞、组织和种子以杀虫剂抗性的组合物和方法。组合物包括编码δ-内毒素和δ-内毒素相关多肽序列的分离核酸分子、包含这些核酸分子的载体和包含这些载体的宿主细胞。组合物还包括内毒素的分离或重组多肽序列、包含这些多肽的组合物以及针对那些多肽的抗体。所述核酸序列可用于DNA构建体或表达盒中，用于生物体中的转化和表达，包括微生物和植物。所述核苷酸或氨基酸序列可以是已被设计成最适于在生物体中表达的合成序列，包括但不局限于，微生物或植物。组合物还包括被转化的细菌、植物、植物细胞、组织和种子。
具体而言，本发明提供了含有编码SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29中所示氨基酸序列的核苷酸序列或SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28中所给出的核苷酸序列的分离核酸分子以及它们的变体和片段。与本发明的核苷酸序列互补或与本发明的序列杂交的核苷酸序列也包括在内。
还提供了可用于产生本发明的多肽的方法，以及利用那些多肽来控制或杀死鳞翅类或鞘翅类害虫的方法。
本发明的组合物和方法可用于产生具有杀虫剂抗性的生物体，尤其是细菌和植物。这些生物体及其来源的组合物是农业用途所需要的。本发明的组合物还可用于产生具有杀虫活性的改变或改良的δ-内毒素或δ-内毒素相关蛋白质，或用于检测产品或生物体中δ-内毒素或δ-内毒素相关蛋白质或者核酸的存在。
附图描述

图1显示了AXMI-004(SEQ ID NO3)与cry1Ac(SEQ ID NO31)、cry1Ca(SEQ ID NO32)、cry2Aa(SEQ ID NO34)、cry3Aa1(SEQ IDNO35)、cry1Ia(SEQ ID NO33)和cry7Aa(SEQ ID NO41)的序列比对。具有C-末端非毒性结构域的毒素如图所示被人为截短。该序列比对显示了以黑色标示的最高度保守氨基酸残基和以灰色标示的高度保守氨基酸残基。发现保守组1是从SEQ ID NO3的大约第174位至大约第196位氨基酸残基。发现保守组2是从SEQ ID NO3的大约第250位至大约第292位氨基酸残基。发现保守组3是从SEQ ID NO3的大约第476位至大约第521位氨基酸残基。发现保守组4是从SEQ ID NO3的大约第542位至大约552位氨基酸残基。发现保守组5是从SEQ IDNO3的大约第618位至大约第628位氨基酸残基。
图2A、B和C显示了AXMI-006(SEQ ID NO7)与cry1Aa(SEQ IDNO30)、cry1Ac(SEQ ID NO31)、cry1Ia(SEQ ID NO33)、cry3Aa1(SEQ ID NO35)、cry3Ba(SEQ ID NO36)、cry4Aa(SEQ ID NO38)、cry6Aa(SEQ ID NO40)、cry7Aa(SEQ ID NO41)、cry8Aa(SEQ IDNO42)、cry10Aa(SEQ ID NO43)、cry16Aa(SEQ ID NO44)、cry19Ba(SEQ ID NO45)和cry24Aa(SEQ ID NO47)的序列比对。具有C-末端非毒性结构域的毒素如图所示被人为截短。该序列比对显示了以黑色标示的最高度保守氨基酸残基和以灰色标示的高度保守氨基酸残基。发现保守组1是从SEQ ID NO7的大约第218位至大约第239位氨基酸残基。发现保守组2是从SEQ ID NO7的大约第300位至大约第350位氨基酸残基。发现保守组3是从SEQ ID NO7的大约第547位至大约第592位氨基酸残基。发现保守组4是从SEQ ID NO7的大约第611位至大约621位氨基酸残基。发现保守组5是从SEQ ID NO7的大约第694位至大约第704位氨基酸残基。
图3A、B和C显示了AXMI-007(SEQ ID NO9)与cry1Aa(SEQ IDNO30)、cry1Ac(SEQ ID NO31)、cry1Ia(SEQ ID NO33)、cry3Aa1(SEQ ID NO35)、cry3Ba(SEQ ID NO36)、cry4Aa(SEQ ID NO38)、cry6Aa(SEQ ID NO40)、cry7Aa(SEQ ID NO41)、cry8Aa(SEQ IDNO42)、cry10Aa(SEQ ID NO43)、cry16Aa(SEQ ID NO44)、cry19Ba(SEQ ID NO45)和cry24Aa(SEQ ID NO47)的序列比对。具有C-末端非毒性结构域的毒素如图所示被人为截短。该序列比对显示了以黑色标示的最高度保守氨基酸残基和以灰色标示的高度保守氨基酸残基。发现保守组1是从SEQ ID NO9的大约第217位至大约第238位氨基酸残基。发现保守组2是从SEQ ID NO9的大约第299位至大约第347位氨基酸残基。发现保守组3是从SEQ ID NO9的大约第445位至大约第590位氨基酸残基。发现保守组4是从SEQ ID NO9的大约第609位至大约619位氨基酸残基。发现保守组5是从SEQ ID NO9的大约第692位至大约第702位氨基酸残基。
图4A、B和C显示了AXMI-008(SEQ ID NO14)与cry1Aa(SEQ IDNO30)、cry1Ac(SEQ ID NO31)、cry1Ia(SEQ ID NO33)、cry2Aa(SBQ ID NO34)、cry3Aa1(SEQ ID NO35)、cry3Bb(SEQ ID NO37)、cry4Aa(SEQ ID NO38)、cry4Ba(SEQ ID NO39)、cry6Aa(SEQ IDNO40)、cry7Aa(SEQ ID NO41)、cry8Aa(SEQ ID NO42)、cry10Aa(SEQ ID NO43)、cry16Aa(SEQ ID NO44)、cry19Ba(SEQ ID NO45)、cry24Aa(SEQ ID NO47)、cry25Aa(SEQ ID NO48)、cry39Aa1(SEQID NO49)和cry40Aa1(SEQ ID NO51)的序列比对。具有C-末端非毒性结构域的毒素如图所示被人为截短。该序列比对显示了以黑色标示的最高度保守氨基酸残基和以灰色标示的高度保守氨基酸残基。发现保守组1是从SEQ ID NO14的大约第185位至大约第206位氨基酸残基。发现保守组2是从SEQ ID NO14的大约第276位至大约第318位氨基酸残基。发现保守组3是从SEQ ID NO14的大约第497位至大约第547位氨基酸残基。发现保守组4是从SEQ ID NO14的大约第576位至大约586位氨基酸残基。发现保守组5是从SEQ ID NO14的大约第657位至大约第667位氨基酸残基。
图5A和B显示了AXMI-008orf2(SEQ ID NO18)与cry19Aa-orf2(SEQ ID NO46)、crybun2-orf2(SEQ ID NO50)、crybun3-orf2(SEQID NO52)、cry4Aa(SEQ ID NO38)和cry4Ba(SEQ ID NO39)的序列比对。该序列比对显示了以黑色标示的最高度保守氨基酸残基和以灰色标示的高度保守氨基酸残基。
图6A、B和C显示了AXMI-009(SEQ ID NO20)与cry1Aa(SEQ IDNO30)、cry1Ac(SEQ ID NO31)、cry1Ca(SEQ ID NO32)、cry1Ia(SEQ ID NO33)、cry3Aa1(SEQ ID NO35)、cry3Ba(SEQ ID NO36)、cry3Bb(SEQ ID NO37)、cry4Aa(SEQ ID NO38)、cry6Aa(SEQ IDNO40)、cry7Aa(SEQ ID NO41)、cry8Aa(SEQ ID NO42)、cry10Aa(SEQ ID NO43)、cry16Aa(SEQ ID NO44)、cry19Ba(SEQ ID NO45)、cry24Aa(SEQ ID NO47)、cry25Aa(SEQ ID NO48)和cry40Aa1(SEQID NO51)的序列比对。具有C-末端非毒性结构域的毒素如图所示被人为截短。该序列比对显示了以黑色标示的最高度保守氨基酸残基和以灰色标示的高度保守氨基酸残基。发现保守组1是从SEQ ID NO20的大约第196位至大约第217位氨基酸残基。发现保守组2是从SEQ IDNO20的大约第269位至大约第311位氨基酸残基。发现保守组3是从SEQ ID NO20的大约第514位至大约第556位氨基酸残基。发现保守组4是从SEQ ID NO20的大约第574位至大约584位氨基酸残基。发现保守组5是从SEQ ID NO20的大约第651位至大约第661位氨基酸残基。
图7A、B和C显示了AXMI-014(SEQ ID NO27)与cry1Aa(SEQ IDNO30)、cry1Ac(SEQ ID NO31)、cry1Ia(SEQ ID NO33)、cry2Aa(SEQ ID NO34)、cry3Aa1(SEQ ID NO35)、cry3Bb(SEQ ID NO37)、cry4Aa(SEQ ID NO38)、cry4Ba(SEQ ID NO39)、cry6Aa(SEQ IDNO40)、cry7Aa(SEQ ID NO41)、cry8Aa(SEQ ID NO42)、cry10Aa(SEQ ID NO43)、cry16Aa(SEQ ID NO44)、cry19Ba(SEQ ID NO45)、cry24Aa(SEQ ID NO47)、cry25Aa(SEQ ID NO48)、cry39Aa1(SEQID NO49)和cry40Aa1(SEQ ID NO51)的序列比对。具有C-末端非毒性结构域的毒素如图所示被人为截短。该序列比对显示了以黑色标示的最高度保守氨基酸残基和以灰色标示的高度保守氨基酸残基。发现保守组1是从SEQ ID NO27的大约第177位至大约第188位氨基酸残基。发现保守组2是从SEQ ID NO27的大约第251位至大约第293位氨基酸残基。发现保守组3是从SEQ ID NO27的大约第483位至大约第533位氨基酸残基。发现保守组4是从SEQ ID NO27的大约第552位至大约562位氨基酸残基。
图8显示了4-20％梯度SDS丙烯酰胺凝胶的照片。泳道1-4包括表达69kD AXMI-004蛋白质的各种浓度的形成孢子的芽孢杆菌细胞培养物。泳道5-8包括各种浓度的BSA。泳道9包含分子量标记物。箭头指示69kD的带。
发明详述本发明针对的是用于调控生物体、尤其是植物或植物细胞中害虫抗性的组合物和方法。所述方法包括用编码本发明δ-内毒素或δ-内毒素相关蛋白质的核苷酸序列转化生物体。具体而言，本发明的核苷酸序列可用于制备具有杀虫活性的植物和微生物。因而提供了已转化的细菌、植物、植物细胞、植物组织和种子。组合物是苏云金芽孢杆菌的δ-内毒素或δ-内毒素相关核酸和蛋白质。所述序列可用于构建表达载体用于随后转化入目的生物体内，作为探针用于分离其它的δ-内毒素或δ-内毒素相关基因，以及用本领域已知方法产生改变的杀虫蛋白质，诸如结构域交换或DNA改组。所述蛋白质可用于抑制或杀死鳞翅类或鞘翅类有害种群以及用于产生具有杀虫活性的组合物。
定义“δ-内毒素”指来自苏云金芽孢杆菌且对一种或多种害虫具有毒性的毒素，所述害虫包括、但不局限于鳞翅目、双翅目和鞘翅目等的种类。在某些情况下，δ-内毒素蛋白质已从其它生物体分离出来了，包括双酶梭状芽孢杆菌(Clostridium bifermentans)和？类芽孢杆菌(Paenibacillus popilliae)。δ-内毒素蛋白质包含由本文所公布的全长核苷酸序列推导出来的氨基酸序列，以及由于使用备选的下游起始位点或者由于产生具有杀虫活性的较短蛋白质的加工所造成的比全长序列更短的氨基酸序列。加工可发生在所述蛋白质所表达的生物体内，或摄取所述蛋白质后在害虫体内。δ-内毒素包括被确认为cry1至cry43、cyt1和cyt2的蛋白质以及Cyt-类毒素。目前有超过250种具有广泛特异性和毒性的已知δ-内毒素。更全面的名单参阅Crickmore等(1998)，Microbiol.Mol.Biol.Rev.62807-813，而定期的更新资料参阅www.biols.susx.ac.uk/Home/Neil-Crickmore/Bt/index上的Crickmore等(2003)″苏云金芽孢杆菌毒素命名(Bacillus thuringiensis toxin nomenclature)″。
细菌基因，诸如本发明的AXMI基因，通常具有多个甲硫氨酸起始密码子接近开发阅读框架的起始点。经常地，开始于一个或多个这些起始密码子的翻译会导致产生功能性蛋白质。这些起始密码子可包括ATG密码子。不过，诸如芽孢杆菌等的细菌还将密码子GTG识别为起始密码子，而翻译起始于GTG密码子的蛋白质在第一个氨基酸处含有甲硫氨酸。此外，并非常常可以争先确定这些密码子中的哪一个在细菌中天然使用。因此，可以理解，利用这些变通性甲硫氨酸密码子之一还可导致产生编码杀虫活性的δ-内毒素蛋白质。例如，本发明AXMI-004δ-内毒素的一个备选起始位点是SEQ ID NO1第385位碱基对处。从此备选起始位点开始的翻译产生了SEQ ID NO5中所显示的氨基酸序列。本发明AXMI-007δ-内毒素蛋白质的一个备选起始位点可以是SEQ ID NO8第151位碱基对处。从此备选起始位点开始的翻译产生了SEQ ID NO11中所显示的氨基酸序列。本发明的AXMI-008δ-内毒素蛋白质的一个备选起始位点可以是SEQ ID NO12的第177位核苷酸处。从此备选起始位点开始的翻译产生了SEQ ID NO16中所示的氨基酸序列。本发明的AXMI-009δ-内毒素蛋白质的一个备选起始位点可以是SEQ ID NO19的第34位核苷酸处。从此备选起始位点开始的翻译产生了SEQ ID NO22中所示的氨基酸序列。本发明的AXMI-009δ-内毒素蛋白质的另一个备选起始位点可以是SEQ ID NO1的第64位核苷酸处。从此备选起始位点开始的翻译产生了SEQ IDNO24中所示的氨基酸序列。本发明的AXMI-014δ-内毒素蛋白质的一个备选起始位点可以是SEQ ID NO25的第136位核苷酸处。从此备选起始位点开始的翻译产生了SEQ ID NO29中所示的氨基酸序列。这些δ-内毒素蛋白质都包括在本发明的范围内，可用于本发明的方法中。
此外，在所公布的编码一个或多个δ-内毒素相关蛋白质的核苷酸序列中可以有一个或多个另外的开发阅读框架。“δ-内毒素相关蛋白质”指用不同于本发明δ-内毒素所用，使用另一开发阅读框架由此处所公布的核苷酸序列编码的蛋白质。诸如此类的蛋白质在本领域中已知可作为辅助蛋白质、稳定序列或δ-内毒素相关蛋白质。这些δ-内毒素相关蛋白质可具有杀虫活性，或可在促进δ-内毒素蛋白质表达中起重要作用。用于检测杀虫活性以及用于测定δ-内毒素相关蛋白质对δ-内毒素蛋白质表达和晶体形成的影响的方法是本领域已知的(参阅，例如，Park等(1999)FEMS Microbiol.Lett.181319-327；Ge等(1998)FEMS Microbiol.Lett.16535-41；Rosso和Delecluse(1997)Appl.Environ.Microbiol.634449-4455)。这些δ-内毒素相关蛋白质包括在本发明范围内，并可单独或与已知的δ-内毒素蛋白质联合用于本文所公布的方法中。在某一实施方案中，δ-内毒素相关蛋白质具有存在于SEQ ID NO18中的氨基酸序列，由SEQ ID NO17的核苷酸序列编码。
“植物细胞”指所有已知形式的植物，包括未分化的组织(如愈伤组织)、悬浮培养细胞、原生质体、叶细胞、根细胞、韧皮部细胞、植物种子、花粉、繁殖体、胚等等。“植物表达盒”指能导致蛋白质在植物细胞中从开放阅读框架表达的DNA构建体。一般其中包含启动子和编码序列。经常地，这样的构建体还包含3’非翻译区。所述构建体可包含“信号序列”或“前导序列”，以促进肽在翻译的同时或翻译后转运至某些细胞内结构，诸如叶绿体(或其它质体)、内质网或高尔基体。
“信号序列”指已知或猜测会导致翻译时或翻译后的跨细胞膜肽转运的序列。在真核细胞中，这通常包括分泌入高尔基体内，具有某些由此产生的糖基化作用。“前导序列”指在翻译时产生的氨基酸序列足以引起肽链共翻译转运至亚细胞器的任何序列。因此，这包括通过进入内质网、穿越液泡、包括叶绿体、线粒体等在内的质体而靶向转运和/或糖基化作用的前导序列。
“植物转化载体”指植物细胞有效转化所必需的DNA分子。这样的分子可由一个或多个植物表达盒组成，且可组装入一个以上的“载体”DNA分子中。例如，二元载体是利用两个非毗连的DNA载体编码植物细胞转化所必需的所有顺式和反式作用功能的植物转化载体(Hellens和Mullineaux(2000)Trends in Plant Science5446-451)。“载体”指设计成在不同宿主细胞之间转移的核酸构建体。“表达载体”指能在外来细胞中掺入、整合并表达异源DNA序列或片段的载体。
“转基因植物”或“已转化的植物”或“稳定转化的植物或细胞组织”指在植物细胞中已掺入或整合了外源核酸序列或DNA片段的植物。这些核酸序列包括外源的或不存在于未转化植物细胞内的那些核酸序列，以及可能是内源的或存在于未转化的植物细胞内的那些核酸序列。“异源的”通常指对于它们所存在的细胞或部分天然基因组而言不是内源性的，且已通过感染、转染、显微注射、电穿孔、显微投影(microprojection)等方式加入细胞内的核酸序列。
“启动子”指起到指导下游编码序列转录的功能的核酸序列。启动子与其它转录和翻译调节核酸序列(也称“调控序列”)一起是目的DNA序列表达所必需的。
本文提供了赋予杀虫活性的新的分离核苷酸序列。还提供了δ-内毒素和δ-内毒素相关蛋白质的氨基酸序列。由此基因翻译所产生的蛋白质可使细胞抑制或杀死摄取它的害虫。
“分离的”或“纯化的”核酸分子或蛋白质或者其生物学活性部分指的是，在用重组技术生产时基本上无其它细胞物质或培养基，或者在化学合成时基本上无化学前体或其它化学物质。优选的，“分离的”核酸中没有天然状态下在该核酸来源的生物体的基因组内处于该核酸两侧(即，位于所述核酸5’和3’末端的序列)的序列(优选蛋白质编码序列)。就本发明的目的而言，“分离的”用于描述核酸分子时，并不包括分离的染色体。例如，在各种实施方案中，分离的编码δ-内毒素或δ-内毒素相关蛋白的核酸分子可包含天然存在于所述核酸来源的细胞的基因组DNA中该核酸分子两侧的小于大约5kb、4kb、3kb、2kb、1kb、0.5kb或0.1kb的核苷酸序列。基本上无细胞物质的δ-内毒素或δ-内毒素相关蛋白质包括具有少于大约30％、20％、10％或5％(干重)的非δ-内毒素或非δ-内毒素相关蛋白质(在此也称为“污染蛋白质”)的蛋白质制品。本发明的各个方面在以下部分有更进一步的详述。
分离的核酸分子及其变体和片段本发明的一方面是关于含有编码δ-内毒素或δ-内毒素相关蛋白质和多肽或其生物学活性部分的核苷酸序列的分离核酸分子，以及足以用作杂交探针来鉴定编码δ-内毒素或δ-内毒素相关蛋白质的核酸分子。用于此处时，术语“核酸分子”意在包括DNA分子(如，cDNA或基因组DNA)和RNA分子(如，mRNA)和用核苷酸类似物产生的所述DNA或RNA的类似物。所述核酸分子可以是单链或双链的，但优选是双链DNA。
编码本发明蛋白质的核苷酸序列包括SEQ ID NOS1、2、4、6、8、10、12、13、15、17、19、21、23、25、26和28中所提出的序列及其互补序列。“互补序列”意指足以与给定核苷酸序列互补、从而可与该给定核苷酸序列杂交形成稳定的双联体的核苷酸序列。由这些核苷酸序列编码的δ-内毒素或δ-内毒素相关蛋白质的相应氨基酸序列如SEQ ID NOS3、5、7、9、11、14、16、18、20、22、24、27和29中所示。
作为这些δ-内毒素或δ-内毒素相关蛋白质编码核苷酸序列的片段的核酸分子也包括在本发明内。“片段”指编码δ-内毒素或δ-内毒素相关蛋白质的核苷酸序列的一部分。核苷酸序列的片段可编码δ-内毒素或δ-内毒素相关蛋白质的生物学活性部分，或可能是可以用下文所述方法作为杂交探针或PCR引物的片段。作为δ-内毒素或δ-内毒素相关核苷酸序列的片段的核酸分子包含至少约15、20、50、75、100、200、300、350、400、450、500、550、600、650、700、750、800、850、900、950、1000、1100、1200、1300、1400、1500、1600、1700、1800、1900、2000、2100、2200、2300、2400、2500、3000、3500、4000、4500、5000、5500个核苷酸，或者多达本文所公布的全长δ-内毒素或δ-内毒素相关蛋白质编码核苷酸序列的核苷酸数目(例如，对于SEQ ID NO1而言是2190个核苷酸，对于SEQ ID NO2而言是1890个核苷酸，等等)，这将取决于目的用途。
本发明的核苷酸序列的片段将编码保留δ-内毒素或δ-内毒素相关蛋白质的生物学活性、从而分别保持了杀虫活性或δ-内毒素相关蛋白质活性的蛋白质片段。“δ-内毒素活性”指杀虫活性。“δ-内毒素相关蛋白质活性”指所述蛋白质具有杀虫活性，或所述蛋白质可提高δ-内毒素蛋白质的表达。这种蛋白质表达的提高可以以任何机制发生。“保持活性”指所述片段具有δ-内毒素或δ-内毒素相关蛋白质的活性的至少约30％、优选至少约50％、更优选至少约70％、还更优选至少约80％。用于测定δ-内毒素相关蛋白质对δ-内毒素蛋白质表达和晶体形成的影响的方法是本领域所已知的(参阅，例如，Park等(1999)FEMS Microbiol.Lett.181319-327；Ge等(1998)FEMSMicrobiol.Lett.16535-41；Rosso和Delecluse(1997)Appl.Environ.Microbiol.634449-4455)。用于测定杀虫活性的方法在本领域内是众所周知的。参阅，例如，Czapla和Lang(1990)J.Econ.Entomol.83(6)2480-2485；Andrews等(1988)Biochem.J.252199-206；Marrone等(1985)J.of Economic Entomology78290-293；和美国专利5,743,477，所有这些都全文收编于此作为参考。
编码本发明蛋白质生物学活性部分的δ-内毒素或δ-内毒素相关蛋白质编码核苷酸序列片段将编码本发明全长δ-内毒素或δ-内毒素相关蛋白质中的至少约15、25、30、50、75、100、125、150、175、200、250、300、350、400、450、500、550、600或650个连续的氨基酸，或最多可达其中所存在的总氨基酸数目(例如，对于SEQ ID NO3而言是629个氨基酸，对于SEQ ID NO5而言是601个氨基酸，等等)。
优选的本发明δ-内毒素或δ-内毒素相关蛋白质是由与SEQ IDNO3、5、7、9、11、14、16、18、20、22、24、27或29中的核苷酸序列足够一致的核苷酸序列编码的。“足够一致的”指用标准参数，通过本文所述序列比对程序之一与参考序列比较而言具有至少约60％或65％序列同一性、优选至少约70％或75％序列同一性、更优选至少约80％或85％序列同一性、最优选至少约90％、91％、92％、93％、94％、95％、96％、97％、98％或99％序列同一性的氨基酸或核苷酸序列。本领域技术人员应认识到，这些数值可适当调整，以确定由于密码子简并性、氨基酸相似性、阅读框架定位等原因而由两种核苷酸序列编码的蛋白质的相应同一性。
为了确定两种氨基酸序列或两种核酸的同一性百分比，将这些序列进行比对以达到最佳的比较效果。两个序列之间的同一性百分比是所述序列共有的相同位置数目的函数(即，同一性百分比＝相同位置的数目/总位置数目(如重叠位置)×100)。在某一实施方案中，所述的两个序列长度相同。两个序列之间的同一性百分比可以用类似于下文所述方法的技术确定，允许或不允许有缺口。在计算同一性百分比时，通常严格匹配的才被计算在内。
两个序列之间同一性百分比的确定可用数学运算法则完成。用于比较两个序列的数学运算法则的非局限性例子是文献Karlin和Altschul(1990)Proc.Natl.Acad.Sci.USA 872264中提出的运算法则，根据文献Karlin和Altschul(1993)Proc.Natl.Acad.Sci.USA 905873-5877进行修正。这样的运算法则被合并入文献Altschul等(1990)J.Mol.Biol.215403的BLASTN和BLASTX程序中。BLAST核苷酸搜索可用BLASTN软件进行(得分＝100，词长＝12)，以获得与本发明δ-内毒素或δ-内毒素相关核酸分子同源的核苷酸序列。BLAST蛋白质搜索可用BLASTN软件进行(得分＝50，词长＝3)，以获得与本发明δ-内毒素或δ-内毒素相关蛋白质分子同源的氨基酸序列。为了获得带缺口的序列比对以进行比较，可如文献Altschul等(1997)Nucleic Acids Res.253389中所述利用Gapped BLAST。或者，可用PSI-Blast进行检测两个分子之间疏远关联的反复搜索。参阅，Altschul等(1997)，同上文。在应用BLAST、Gapped BLAST和PSI-Blast程序时，可使用各个程序(如，BLASTX和BLASTN)默认的参数。见www.ncbi.nlm.nih.gov。另一用于比较序列的数学运算法则的非限制性例子是ClustalW运算法则(Higgins等(1994)Nucleic AcidsRes.224673-4680)。ClustalW可比较序列，并将氨基酸或DNA序列全部比对，因此可提供有关整个氨基酸序列的序列保守性的资料。ClustalW运算法则被用于数个商品化的DNA/氨基酸分析软件包中，诸如载体NTi Program Suite(Informax，Inc)的ALIGNX模块。用ClustalW进行氨基酸序列的比对后，可评估氨基酸同一性百分比。可用于ClustalW序列比对分析的软件程序的非局限性例子是GeneDocTM。GenedocTM(Karl Nicholas)可评估多个蛋白质之间的氨基酸(或DNA)相似性和同一性。用于序列比较的数学运算法则的另一非限制性例子是文献Myers和Miller(1988)CABIOS 411-17中的运算法则。这样的运算法则被引入ALIGN程序(版本2.0)中，它是GCG序列比对软件包的一部分(可获自Accelrys，Inc.，9865Scranton Rd.，San Diego，California，USA)。当应用ALIGN程序来比较氨基酸序列时，可使用PAM120加权残基表、缺口长度罚分12和缺口罚分4。
本发明还包括变体核酸分子。δ-内毒素或δ-内毒素相关蛋白质编码核苷酸序列的“变体”包括编码本文所公布的δ-内毒素或δ-内毒素相关蛋白质、但由于遗传密码简并性而有保守性差别的序列，以及如上所述足够相似的那些序列。天然存在的等位基因变体可用众所周知的分子生物学技术鉴定，诸如聚合酶链式反应(PCR)和下文概述的杂交技术。变体核苷酸序列还包括合成来源的核苷酸序列，例如如下所述通过定位诱变产生、但仍然编码本发明公布的δ-内毒素或δ-内毒素相关蛋白质的序列。包含在本发明范围内的变体蛋白质是有生物学活性的，也就是说，它们继续具有天然蛋白质的目的生物学活性，即保持杀虫活性。“保持活性”是指变体具有δ-内毒素或δ-内毒素相关蛋白质的活性的至少约30％、优选至少约50％、更优选至少约70％、还更优选至少约80％。用于测量杀虫活性的方法在本领域中是众所周知的。参阅，例如，Czapla和Lang(1990)L.Econ.Entomol.83(6)2480-2485；Andrews等(1988)Biochem.J.252199-206；Marrone等(1985)J.of Economic Entomology78290-293；以及美国专利5,743,477，所有这些均全部收编于此作为参考。
本发明还包括变体核酸分子。δ-内毒素或δ-内毒素相关编码核苷酸序列的“变体”包括那些编码本文所公布的δ-内毒素或δ-内毒素相关蛋白质、但由于遗传密码简并性而有保守性差异的序列以及如上所述充分相同的那些序列。天然存在的等位基因变体可用众所周知的分子生物学技术鉴定，诸如聚合酶链式反应(PCR)和下述杂交技术。变体核苷酸序列还包括合成来源的核苷酸序列，例如如下所述通过定位诱变产生、但仍然编码本发明公布的δ-内毒素或δ-内毒素相关蛋白质的序列。
技术熟练人员应认识到，可通过突变在本发明的核苷酸序列中引入变异，从而导致所编码的δ-内毒素或δ-内毒素相关蛋白质的氨基酸序列中的变化，但不改变所述蛋白质的生物学活性。因此，可通过在本文所公布的相应核苷酸序列中引入一个或多个核苷酸取代、添加或缺失而制备变异的分离核酸分子，从而在所编码的蛋白质中引入一个或多个氨基酸取代、添加或缺失。突变可通过标准技术引入，诸如定位诱变和PCR介导诱变。这样的变异核苷酸序列也包括在本发明的范围内。
例如，优选的，可以在一个或多个预知的、优选非必需氨基酸残基处进行保守氨基酸取代。“非必需的”氨基酸残基是可以在δ-内毒素或δ-内毒素相关蛋白质序列的野生型序列中发生改变、而不改变生物学活性的残基，而“必需的”氨基酸残基是生物学活性所必需的。“保守的氨基酸取代”是用具有相似侧链的氨基酸残基替换某一氨基酸残基。具有相似侧链的氨基酸残基家族在本领域中已有定义。这些家族包括具有碱性侧链的氨基酸(如，赖氨酸、精氨酸、组氨酸)、具有酸性侧链的氨基酸(如，天冬氨酸、谷氨酸)、具有不带电荷的极性侧链的氨基酸(如，甘氨酸、天冬酰胺、谷氨酰胺、丝氨酸、苏氨酸、酪氨酸、半胱氨酸)、具有非极性侧链的氨基酸(如，丙氨酸、缬氨酸、亮氨酸、异亮氨酸、脯氨酸、苯丙氨酸、甲硫氨酸、色氨酸)、具有β-分支侧链的氨基酸(如，苏氨酸、缬氨酸、异亮氨酸)和具有芳香族侧链的氨基酸(如，酪氨酸、苯丙氨酸、色氨酸、组氨酸)。
在δ-内毒素蛋白质之间通常有5个高度保守的区域，主要集中在结构域的中心或结构域之间的连接处(Rajamohan等(1998)Prog.Nucleic Acid Res.Mol.Biol.601-23)。多种δ-内毒素的保守氨基酸块以及共有序列可参阅以下文献Schnepf等(1998)Microbio.Mol.Biol.Rev.62775-806和Lereclus等(1989)，编码苏云金芽孢杆菌类芽孢δ-内毒素的基因的作用、结构和分子组成(Role，Structure，and Molecular Organization of the Genes Coding forthe Parasporal d-endotoxins of Bacillus thuringiensis)。在“原核发育的调控(Regulation of Procaryotic Development)”一书中，Issar Smit，Slepecky，R.A.，Setlow，P.美国微生物学会，Washington，D.C.20006。已提出具有这些保守区的δ-内毒素可共享相似的结构，由三个结构域组成(Li等(1991)Nature 353815-821)。结构域I在δ-内毒素之间具有最高的相似性(Bravo(1997)J.Bacteriol.1792793-2801)。
可在非保守区内进行保持活性的氨基酸取代。一般而言，不对保守氨基酸残基、或定位在保守基元内的氨基酸残基进行这样的取代，其中所述的残基对于蛋白质活性是必需的。保守的、并且可能为蛋白质活性所必需的残基例子包括，例如，在所提供的序列比对中含有的所有蛋白质之间均相同的残基。保守的、但可能允许发生保守性氨基酸取代而仍保持活性的残基的例子包括例如在所提供的序列比对中所包括的所有蛋白质之间仅有保守性取代的残基。不过，本领域技术人员应理解，功能性变体可在保守残基中具有微小的保守或非保守变异。
或者，可通过诸如饱和诱变等方法沿着所有或部分编码序列随机引入突变而制备变异核苷酸序列，并可对所产生的突变体筛选赋予δ-内毒素或δ-内毒素相关活性的能力，以鉴别保持活性的突变体。诱变后，所编码的蛋白质可重组表达，而蛋白质的活性可用标准检测技术测定。
用诸如PCR、杂交等方法可鉴定相应的δ-内毒素或δ-内毒素相关序列，这样的序列与本发明的序列基本上相同。参阅，例如，SambrookJ.和Russell，D.W.(2001)分子克隆实验室手册MolecularCloningA Laboratory Manual.(冷泉港实验室出版社，冷泉港，纽约)和Innis等(1990)PCR流程方法和应用指南PCR ProtocolsA Guide to Methods and Applications(Academic Press，NY)。
在杂交方法中，可用全部或部分δ-内毒素或δ-内毒素相关核苷酸序列筛选cDNA或基因组文库。所述cDNA和基因组文库的构建方法是本领域普遍已知的，并被描述在Sambrook和Russell，2001中。所谓的杂交探针可以是基因组DNA片段、cDNA片段、RNA片段或其它寡核苷酸，它们可用诸如32P等可检测基团或诸如其它放射性同位素、荧光化合物、酶或酶辅因子等任何其它可检测标记物进行标记。可通过标记基于本文所公布的已知δ-内毒素或δ-内毒素相关编码核苷酸序列的合成寡核苷酸制备用于杂交的探针。此外，还可使用基于核苷酸序列或所编码的氨基酸序列中的保守核苷酸或氨基酸残基设计的简并引物。所述探针通常包含在严谨条件下与本发明δ-内毒素或δ-内毒素相关编码核苷酸序列或其片段或变体的至少约12个、优选约25个、更优选至少约50、75、100、125、150、175、200、250、300、350或400个连续核苷酸杂交的核苷酸序列区。用于杂交的探针的制备是本领域所普遍已知的，并且公开在此处收编作为参考的文献Sambrook和Russell，2001中。
在杂交技术中，已知的核苷酸序列的全部或部分被用作与来自选定生物体的克隆基因组DNA片段或cDNA片段群(即，基因组或cDNA文库)中存在的其它相应核苷酸序列选择性杂交的探针。杂交探针可以是基因组DNA片段、cDNA片段、RNA片段或其它寡核苷酸，且可用诸如32P等可检测基团或任何其它可检测标记物标记。因此，例如，用于杂交的探针可通过标记基于本发明的δ-内毒素或δ-内毒素相关序列的合成寡核苷酸而制备。杂交探针的制备方法以及cDNA和基因组文库的构建方法是本领域通常已知的，并公开在Sambrook等(1989)分子克隆实验室手册Molecular CloningA Laboratory Manual(第二版，冷泉港实验室出版社，Plainview，纽约)中。
例如，本文所公布的整个δ-内毒素或δ-内毒素相关序列或其一个或多个部分可用作能特异性地与相应δ-内毒素或δ-内毒素相关样序列以及信使RNA杂交的探针。为了实现各种条件下的特异杂交，所述探针包含独特的序列，并且优选长度为至少约10个核苷酸、最优选至少约20个核苷酸。这样的探针可用于通过PCR从选定的生物体中扩增相应的δ-内毒素或δ-内毒素相关序列。这一技术可用于从目的生物体中分离额外的编码序列，或作为诊断检测法用于确定生物体中编码序列的存在与否。杂交技术包括铺板的DNA文库(或者噬斑或者菌落，参阅，例如，Sambrook等(1989)分子克隆实验室手册(第二版，冷泉港实验室出版社，Plainview，纽约))的杂交筛选。
这些序列的杂交可在严谨条件下进行。“严谨条件”或“严谨的杂交条件”指，在此条件下，探针与其靶序列的杂交程度可检测性地高于其它序列(例如，至少高于背景2倍)。严谨条件是随序列而定的，在不同的环境下有所不同。通过控制杂交和/或漂洗条件的严谨性，可鉴别与探针100％互补的靶序列(同源探测)。或者，可将严谨条件调整成允许序列中有一些错配，从而检测较低程度的相似性(异源探测)。通常，探针的长度小于约1000个核苷酸，优选长度小于500个核苷酸。
通常，严谨条件是这样的条件，其中在pH7.0至8.3时盐浓度小于约1.5M Na离子、通常约0.01至1.0M Na离子浓度(或其它盐)，并且对于短探针(如10-50个核苷酸)而言温度至少约30℃，而对于长探针(例如，大于50个核苷酸)而言至少约60℃。还可通过添加诸如甲酰胺等去稳定剂来达到严谨条件。示范性的低严谨条件包括用30-35％甲酰胺、1M NaCl、1％SDS(十二烷基硫酸钠)的缓冲液在37℃进行杂交，在1X至2X SSC(20X SSC＝3.0M NaCl/0.3M柠檬酸三钠)在50-55℃漂洗。示范性的中等严谨条件包括在40-45％甲酰胺、1.0M NaCl、1％SDS中于37℃杂交，并在0.5X至1X SSC于55-60℃漂洗。示范性的高严谨条件包括在50％甲酰胺、1M NaCl、1％SDS中于37℃进行杂交，并在0.1X SSC中于60-65℃漂洗。任选的，漂洗缓冲液中可包含约0.1％至约1％SDS。杂交的持续时间一般少于大约24小时，通常约为4至12小时。
一般而言，特异性是杂交后漂洗条件的函数，关键因素是最终漂洗溶液的离子强度和温度。对于DNA-DNA杂交，Tm可用Meinkoth等Wahl(1984)Anal.Biochem.138267-284中的公式估计Tm＝81.5℃+16.6(log M)+0.41(％GC)-0.61(％form)-500/L；其中的M是单价阳离子的摩尔浓度，％GC是DNA中鸟嘌呤和胞嘧啶核苷酸的百分比，％form是杂交溶液中甲酰胺的百分比，而L是碱基对中杂交体的长度。Tm是互补靶序列的50％与完全匹配的探针杂交的温度(在确定的离子强度和pH下)。对于每1％的错配，Tm下降约1℃；因此，为了与所期望的同一性的序列杂交，可以调节Tm、杂交和/或漂洗条件。例如，如果搜寻具有≥90％同一性的序列，Tm可下降10℃。通常，将严谨条件选择成比该具体序列与其互补序列在确定的离子强度和pH的热融点(Tm)低约5℃。不过，严格的严谨条件可利用比热融点(Tm)低1、2、3或4℃的条件进行杂交和/或漂洗；中等的严谨条件可利用比热融点(Tm)低6、7、8、9或10℃的条件进行杂交和/或漂洗；低严谨条件可利用可利用比热融点(Tm)低11、12、13、14、15或20℃的条件进行杂交和/或漂洗。利用所述公式、杂交和漂洗组合物以及预期的Tm，普通技术人员可理解杂交和/或漂洗溶液的严谨性的变化已得到了本质的描述。如果所需程度的错配导致Tm小于45℃(水溶液)或32℃(甲酰胺溶液)，则优选提高SSC浓度，以利用较高的温度。有关核酸杂交的详尽指南可参阅文献Tijssen(1993)生化和分子生物学实验技术-用核酸探针进行杂交(Laboratory Techniques inBiochemistry and Molecular Biology-Hybridization with NucleicAcid Probes)，第I部分，第二章(Elsevier，纽约)；和Ausubel等编辑(1995)分子生物学通用方法(Current Protocols inMolecular Biology)，第二章(Greene Publishing andWiley-Interscience，New York)。参阅Sambrook等(1989)分子克隆实验室手册(Molecular CloningA Laboratory Manual)(第二版，冷泉港实验室出版社，Plainview，New York)。
分离的蛋白质及其变体和片段
δ-内毒素和δ-内毒素相关蛋白质也包含在本发明的范围内。“δ-内毒素蛋白质”指具有SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29中所示氨基酸序列的蛋白质。“δ-内毒素相关蛋白质”指具有SEQ ID NO18中所示氨基酸序列的蛋白质。同时还提供了它们的片段、生物学活性部分和变体，它们可用于实践本发明的方法。“片段”或“生物学活性部分”包括含有编码如SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29中所示δ-内毒素或δ-内毒素相关蛋白质的氨基酸序列的一部分且保持δ-内毒素活性或δ-内毒素相关活性的多肽片段。δ-内毒素或δ-内毒素相关蛋白质的生物学活性部分可以是例如10、25、50、100或更多个氨基酸长的多肽。这样的生物学活性部分可通过重组技术制备，并评估其δ-内毒素或δ-内毒素相关活性。检测杀虫活性的方法是本领域众所周知的。参阅，例如，Czapla和Lang(1990)J.Econ.Entomol.83(6)2480-2485；Andrews等(1988)Biochem.J.252199-206；Marrone等(1985)J.of Economic Entomology 78290-293；以及美国专利5,743,477，所有这些均全部收编于此作为参考。用于此处时，片段含有SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29中的至少8个毗连氨基酸。不过，本发明还包含其它的片段，诸如该蛋白质中的任何大于约10、20、30、50、100、150、200、250、300、350、400、450、500、550、600和650个氨基酸的片段。
“变体”指具有与SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示氨基酸序列至少约60％、65％、优选至少约70％、75％、更优选至少约80％、85％、最优选至少约90％、91％、92％、93％、94％、95％、96％、97％、98％或99％相同的氨基酸序列的蛋白质或多肽。变体还包括在严谨条件下与SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28的核酸分子或其互补分子杂交的核酸分子编码的多肽。这样的变体通常保持δ-内毒素或δ-内毒素相关活性。变体包括由于诱变而氨基酸序列有所区别的多肽。本发明所包含的变体蛋白质是有生物学活性的，即，它们继续具有天然蛋白质的所需生物学活性，即保持杀虫活性。检测杀虫活性的方法是本领域众所周知的。参阅，例如，Czapla和Lang(1990)J.Econ.Entomol.83(6)2480-2485；Andrews等(1988)Biochem.J.252199-206；Marrone等(1985)J.of Economic Entomology 78290-293；以及美国专利5,743,477，所有这些均全部收编于此作为参考。
改变或改良的变体已认识到可用各种方法改变δ-内毒素或δ-内毒素相关蛋白质的DNA序列，并且这些改变可能产生所编码的蛋白质氨基酸序列不同于本发明δ-内毒素或δ-内毒素相关蛋白质氨基酸序列的DNA序列。这一蛋白质可以多种方式改变，包括氨基酸取代、缺失、截短和插入。这些操作方法是本领域通常已知的。例如，δ-内毒素或δ-内毒素相关蛋白质的氨基酸序列变体可通过DNA中的突变来制备。这还可以通过数种诱变形式之一和/或定向进化来完成。在某些方面，氨基酸序列中编码的改变基本上不会影响蛋白质的功能。这样的变体将具有所需的杀虫活性。不过，应理解δ-内毒素或δ-内毒素相关蛋白质赋予杀虫活性的能力可基于本发明组合物通过使用所述技术得以提高。例如，可以在于DNA复制期间呈现出高比率碱基错误掺入的宿主细胞中表达δ-内毒素或δ-内毒素相关蛋白质，诸如XL-1 Red(Stratagene)。在所述株系中增殖后，可分离δ-内毒素或δ-内毒素相关DNA(例如通过制备质粒，或通过PCR扩增并将产生的PCR片段克隆入载体中)，在非诱变性菌株中培养δ-内毒素或δ-内毒素相关突变，鉴定具有杀虫活性的突变的δ-内毒素或δ-内毒素相关基因，例如进行检测来测试杀虫活性。通常，将所述蛋白质混和并用于喂服试验中。参阅，例如，Marrone等(1985)J.of Economic Entomology78290-293。这样的试验可包括使植物接触一种或多种害虫并测定植物存活的能力和/或引起害虫死亡的能力。导致毒性增强的突变的例子参阅Schnepf等(1998)Microbiol.Mol.Biol.Rev.62775-806。
或者，可在氨基或羧基末端对许多蛋白质的序列进行改变而基本上不影响其活性。这可包括利用现代分子方法引入的插入、删除或改变，诸如PCR，包括由于在PCR扩增所用的寡核苷酸中含有氨基酸编码序列而改变或延长蛋白质编码序列的PCR扩增。或者，所加入的蛋白质序列可包括完整的蛋白质编码序列，诸如那些在本领域中通常用于产生蛋白质融合体的序列。这样的融合蛋白质常常用于(1)增加目的蛋白质的表达，(2)引入结合结构域、酶促活性或表位以方便蛋白质纯化、蛋白质检测或本领域已知的其它实验性应用，(3)使蛋白质的分泌或翻译定向于亚细胞性细胞器，诸如革兰氏阴性细菌的周质空间，或真核细胞的内质网，后者常常导致蛋白质的糖基化。
本发明的变体核苷酸和氨基酸序列还包括来自诸如DNA改组等诱变和重组方法的序列。通过这样的步骤，可利用一个或多个不同的δ-内毒素或δ-内毒素相关蛋白质编码区构建具有所需特性的新的δ-内毒素或δ-内毒素相关蛋白质。以此方式，由含有具有充分的序列同一性且可体外或体内同源重组的序列区的相关序列多核苷酸群产生重组多核苷酸文库。例如，用此方法，可在本发明的δ-内毒素或δ-内毒素相关基因与其它已知δ-内毒素或δ-内毒素相关基因之间对编码目的结构域的序列基元进行改组，以获得编码具有改良目的特性的蛋白质的新基因，诸如提高杀虫活性。所述的DNA改组策略是本领域所已知的。参阅，例如，Stemmer(1994)Proc.Natl.Acad.Sci.USA9110747-10751；Stemmer(1994)Nature 370389-391；Crameri等(1997)Nature Biotech.15436-438；Moore等(1997)J.Mol.Biol.272336-347；Zhang等(1997)Proc.Natl.Acad.Sci.USA944504-4509；Crameri等(1998)Nature 391288-291；以及美国专利5,605,793和5,837,458。
结构域交换或改组是产生改变的δ-内毒素或δ-内毒素相关蛋白质的另一机制。结构域II和III可在δ-内毒素蛋白质之间进行交换，产生具有改良的杀虫活性或目标谱的杂合或嵌合毒素。产生重组蛋白质并检测其杀虫活性的方法在本领域中是众所周知的(参阅，例如，Naimov等(2001)Appl.Environ.Microbiol.675328-5330；de Maagd等(1996)Appl.Environ.Microbiol.621537-1543；Ge等(1991)J.Biol.Chem.26617954-17958；Schnepf等(1990)j.Biol.Chem.26520923-20930；Rang等91999)Appl.Environ.Micriobiol.652918-2925)。
植物转化可通过本领域已知的数种技术之一完成植物细胞的转化。首先，以允许其在植物细胞中表达的方式改造δ-内毒素或δ-内毒素相关基因。通常表达这种蛋白质的构建体应含有驱使基因转录的启动子以及允许转录终止和聚腺苷酸化的3’非翻译区。这样的构建体的组成在本领域中是众所周知的。在某些情况下，可有效改造基因，使得产生的肽被分泌出来，或定向于植物细胞中。例如，可改造基因，使其含有信号肽，以方便所述的肽转移至内质网。还可能优选的是，改造植物表达盒使其含有内含子，从而使内含子的mRNA加工为表达所必需。
通常这一“植物表达盒”将被插入“植物转化载体”中。该植物转化载体可由完成植物转化所需的一个或多个DNA载体组成。例如，本领域的通用惯例是利用由一个以上毗连DNA区段组成的植物转化载体。这些载体在本领域中常常称为“二元载体”。二元载体以及具有辅助质粒的载体是最常用于农杆菌介导的转化中的，其中完成有效转化所需的DNA区段的大小和复杂性是相当大的，并且将功能分摊到分开的DNA分子上比较有利。二元载体通常包含含有T-DNA转移所必需的顺式作用序列(诸如左边界和右边界)、经改造能在植物细胞中表达的可选择标记物以及“目的基因”(经改造能在期望产生转基因植物的植物细胞中表达的基因)的质粒载体。还存在于此质粒载体中的是细菌复制所必需的序列。顺式作用序列以可有效转移入植物细胞并在其中表达的形式安排。例如，可选择标记基因和目的基因定位于左和右边界之间。常常有另一个质粒载体包含介导T-DNA从农杆菌转移至植物细胞的的反式作用因子。按照本领域中所理解的，此质粒常常具有使植物细胞可被农杆菌感染并通过在边界序列处的断裂以及病毒介导的DNA转移来转移DNA的致病作用(Vir基因)(Hellens和Mullineaux(2000)Trends in Plant Science，5446-451)。数种类型的农杆菌菌株(如LBA4404、GV3101、EHA101、EHA105等)可用于植物转化。该第二种质粒载体不是用诸如显微投影、显微注射、电穿孔、聚乙二醇等其它方法转化植物所必需的。
一般而言，植物转化方法包括将异源DNA转移入靶植物细胞(如，不成熟或成熟的胚、悬浮培养物、未分化的愈伤组织、原生质体，等等)中，随后进行最大极限水平的适当选择(取决于可选择标记基因)，以从未转化细胞集合组中回收转化的植物细胞。通常将外植体转移至新鲜置换的同种培养基中并进行常规培养。随后，在放置于补充了最大极限水平的选择剂的再生培养基中后已转化的细胞被分化成芽。然后将所述的芽转移到选择性生根培养基中以回收生根的芽或苗。然后将转基因苗培养成成熟的植物并产生能繁殖的种子(如Hiei等(1994)The Plant Journal 6271-282；Ishida等(1996)NatureBiotechnology 14745-750)。通常将外植体转移至新鲜补充的同种培养基中并进行常规培养。产生转基因幼苗的有关技术和方法的全面描述参阅文献Ayres和Park，1994(Critical Reviews in PlantScience 13219-239)以及Bommineni和Jauhar，1997(Maydica 42107-120)。由于被转化的物质包含许多细胞，转化和未转化的细胞都存在于任何受试靶愈伤片或组织或细胞组中。杀死未转化细胞并允许转化细胞增殖的能力使被转化的植物培养物得以生长。通常，去除未转化细胞的能力是对快速回收被转化植物细胞并成功产生转基因植物的限制。
转基因植物的产生可用数种方法之一完成，包括但不局限于通过农杆菌将异源DNA引入植物细胞中(农杆菌介导的转化)、用粘附于颗粒上的外源性外来DNA轰击植物细胞以及各种其它非粒子定向介导的方法(如Hiei等(1994)The Plant Journal 6271-282；Ishida等(1996)Nature Biotechnology 14745-750；Ayres和Park(1994)Critical Reviews in Plant Science 13219-239；Bommineni和Jauhar(1997)Maydica 42107-120)转移DNA。
转化方法以及将核苷酸序列引入植物的方法可能随着待转化的植物或植物细胞的类型(即单子叶或双子叶植物)而变化。将核苷酸序列引入植物细胞并随后插入植物基因组中的合适方法包括显微注射(Crossway等(1986)Biotechniques 4320-334)、电穿孔(Riggs等(1986)Proc.Natl.Acad.Sci.USA 835602-5606)、农杆菌介导的转化(美国专利5,563,055；美国专利5,981,840)、直接基因转移(Paszkowski等(1984)EMBO J.32717-2722)和弹道颗粒加速(参阅，例如，美国专利4,945,050；美国专利5,879,918；美国专利5,886,244；美国专利5,932,782；Tomes等(1995)″通过显微投射轰击将DNA定向转移入完整的植物细胞中(Direct DNA Transfer into Intact PlantCells via Microprojectile Bombardment)″，在“植物细胞、组织和器官培养基本方法(Plant Cell，Tissue，and Organ CultureFundamental Methods)”一书中，Gamborg和Phillips编辑(Springer-Verlag，Berlin)；McCabe等(1988)Biotechnology6923-926)、气溶胶束转化(美国已出版专利申请20010026941；美国专利4,945,050；国际专利申请WO91/00915；美国已出版专利申请2002015066)以及Lec1转化(WO00/28058)。也可参阅Weissinger等(1988)Ann.Rev.Genet.22421-477；Sanford等(1987)微粒科学和技术(Particulate Science and Technology)527-37；Christou等(1988)Plant Physiol.87671-674；McCabe等(1988)Bio/Technology 6923-926；Finer和McMullen(1991)In Vitro CellDev.Biol.27P175-182；Singh等(1998)Theor.Appl.Genet.96319-324(大豆)；Datta等(1990)Biotechnology 8736-740；Klein等(1988)Proc.Natl.Acad.Sci.USA854305-4309；美国专利5,240,855；美国专利5,322,783和5,324,646；Tomes等(1995)″通过微粒轰击将DNA定向转移入完整的植物细胞中(Direct DNATransfer into Intact Plant Cells via MicroprojectileBombardment)″，在“植物细胞、组织和器官培养基本方法(PlantCell，Tissue，and Organ CultureFundamental Methods)”一书中，编辑Gamborg和Phillips(Springer-Verlag，Berlin)；Klein等(1988)Plant Physiol.91440-444；Hooykaas-Van Slogteren等(1984)Nature(London)311763-764；美国专利5,736,369；Bytebier等(1987)Proc.Natl.Acad.Sci.USA845345-5349(Liliaceae)；De Wet等(1985)在“胚珠组织的实验操作(The ExperimentalManipulation of Ovule Tissues)”一书中，编辑Chapman等(Longman，New York)，第197-209页；Kaeppler等(1990)植物细胞报告PlantCell Reports 9415-418和Kaeppler等(1992)Theor.Appl.Genet.84560-566；D′Halluin等(1992)Plant Cell 41495-1505；Li等(1993)Plant Cell Reports 12250-255以及Christou和Ford(1995)Annals of Botany 75407-413；Osjoda等(1996) NatureBiotechnology 14745-750，所有这些均收编于此作为参考。
在将外源性外来DNA整合入植物细胞中后，在培养基中进行最大极限水平的适当选择，以杀死未转化的细胞，并通过定期转移至新鲜培养基中而分离和增殖在此选择处理中存活的据推定已转化的细胞。通过连续传代并进行适当的选择，可识别并增殖被所述质粒载体转化的细胞。然后用分子和生化方法证实转基因植物基因组中被整合的异源目的基因的存在。
已被转化的细胞可以传统方式生长成植物。参阅，例如McCormick等(1986)Plant Cell Reports 581-84。然后可培养这些植物，用同样的被转化株系或不同的株系授粉，并鉴定具有所需表型特征的组成型表达的所得杂合体。可培养两代或多代，以保证所需表型特征的表达稳定维持和遗传，然后收获种子以保证已实现了所需表型特征的表达。以此方式，本发明提供了在其基因组中稳定掺入了本发明的核苷酸构建体、例如本发明的表达盒的已转化种子(也称为“转基因种子”)。
本发明的δ-内毒素或δ-内毒素相关序列可存在于用于在目的植物中表达的表达盒中。所述的盒将包括可操作性的与本发明序列连接的5′和3′调控序列。“可操作性的连接”指在启动子和第二序列之间的功能性联接，其中的启动子序列起始并介导了与第二序列相应的DNA序列的转录。通常，可操作性连接意味着被连接的核酸序列是毗邻的，并且，在需连接两个蛋白质编码区时，则是毗邻且处于同一阅读框架中。所述的盒可另外含有至少一个其它基因以共转化入生物体中。或者，所述的其它基因可存在于多个表达盒中。
向所述的表达盒中提供多个限制性位点，以供插入δ-内毒素或δ-内毒素相关序列，处于调控区的转录调控下。
所述的表达盒沿5′-3′转录方向将包含在植物中发挥作用的转录和翻译起始区(即，启动子)、本发明的DNA序列以及转录和翻译终止区(即终止区)。启动子对于植物宿主和/或本发明的DNA序列而言可能是天然的或类似的、或者外源的或异源的。此外，启动子可以是天然序列或可以是合成序列。启动子对于植物宿主而言是“天然的”或“同源的”，指的是启动子在该启动子将被引入的天然植物中存在。启动子对于本发明DNA序列而言是“外源的”或“异源的”，指的是启动子对于可操作性连接的本发明DNA序列而言不是天然或自然存在的启动子。
终止区可以是对于转录起始区而言是天然的，可以是就可操作性连接的目的DNA序列而言是天然的，可以是就植物宿主而言是天然的，或者可以来自其它来源(即，对于启动子、目的DNA序列、植物宿主或它们的任何组合而言是外源的或异源的)。方便的终止区可从根癌农杆菌的Ti-质粒获得，诸如章鱼碱合酶和胭脂碱合酶终止区。还可参阅Guerineau等(1991)Mol.Gen.Genet.262141-144；Proudfoot(1991)Cell 64671-674；Sanfacon等(1991)Genes Dev.5141-149；Mogen等(1990)Plant Cell 21261-1272；Munroe等(1990)Gene91151-158；Ballas等(1989)Nucleic Acids Res.177891-7903；以及Joshi等(1987)Nucleic Acid Res.159627-9639。
在适当的情况下，可将基因最优化以提高其在被转化宿主细胞中的表达。即，所述基因可以用宿主细胞偏好的密码子合成以促进表达，或者可以宿主偏好的密码子使用频率用密码子合成。例如，有关宿主偏好的密码子使用的讨论参阅Campbell和Gowri(1990)PlantPhysiol.921-11。合成植物偏好基因的方法是本领域所已知的。参阅，例如，美国专利6,320,100、6,075,185、5,380,831和5,436,391，美国已出版专利申请20040005600和20010003849，以及Murray等(1989)Nucleic Acids Res.17477-498，在此收编作为参考。
在一个实施方案中，目的核酸被定向于叶绿体中表达。以此方式，当目的核酸不是直接插入叶绿体的情况下，表达盒将另外含有编码转运肽的核酸以指导目的基因产物进入叶绿体。这样的转运肽在本领域中是已知的。参阅，例如，Von Heijne等(1991)Plant Mol.Biol.Rep.9104-126；Clark等(1989)J.Biol.Chem.26417544-17550；Della-Cioppa等(1987)Plant Physiol.84965-968；Romer等(1993)Biochem.Biophys.Res.Commun.1961414-1421；以及Shah等(1986)Science 233478-481。
用于叶绿体转化的方法在本领域中是已知的。参阅，例如，Svab等(1990)Proc.Natl.Acad.Sci.USA 878526-8530；Svab和Maliga(1993)Proc.Natl.Acad.Sci.USA 90913-917；Svab和Maliga(1993)EMBO J.12601-606。所述方法依赖于含可选择标记的DNA的粒子枪投递以及通过同源重组将DNA定向于质体基因组。此外，质体转化可通过用核编码及质体指导的RNA聚合酶的组织优选表达而反式激活沉默的质体携带的转基因而完成。这样的体系已被报道于文献McBride等(1994)Proc.Natl.Acad.Sci.USA 917301-7305中。
可优化定向于叶绿体的目的核酸以表达于叶绿体中，从而解决植物细胞核和此细胞器之间的密码子使用中的差异。以此方式，目的核酸可以用叶绿体偏好的密码子合成。参阅，例如，美国专利5,380,831，在此收编作为参考。
植物转化的评估将异源性的外来DNA引入植物细胞中后，用各种方法证实植物基因组中异源基因的转化或整合，诸如与被整合基因相关的核酸、蛋白质和代谢物的分析。
PCR分析PCR分析是在移植至土壤中之前的较早阶段筛选已转化的细胞、组织或芽是否存在被掺入基因的快速方法(Sambrook和Russell，2001)。PCR是用特异于目的基因或农杆菌载体背景等的寡核苷酸引物进行的。
DNA杂交分析植物转化通过基因组DNA的DNA印迹分析证实(Sambrook和Russell，2001)。通常，从转化株中提取总DNA，用适当的限制酶消化，在琼脂糖凝胶中进行分级分离并转移至硝酸纤维素或尼龙膜上。然后依照标准技术，用例如放射性标记了32P的靶DNA片段探测所述的膜或“印迹”，以证实被导入基因在植物基因组中的整合(Sambrook和Russell，2001.分子克隆实验室手册MolecularCloningA Laboratory Manual.冷泉港实验室出版社，冷泉港，纽约)。
RNA杂交分析依照本领域中常规使用的标准方法，从转化株的特异组织中分离RNA，在甲醛琼脂糖凝胶中分级分离，印迹至尼龙滤膜上(Sambrook，J.和Russell，D.W.2001.分子克隆实验室手册Molecular CloningA Laboratory Manual.冷泉港实验室出版社，冷泉港，纽约)。然后用本领域已知方法(Sambrook和Russell，2001)，通过将所述滤膜与来自δ-内毒素或δ-内毒素相关蛋白质的放射性探针杂交而测试δ-内毒素或与δ-内毒素相关基因编码的RNA的表达。
蛋白质印迹和生化检测可通过标准步骤(Sambrook，J.和Russell，D.W.2001.分子克隆实验室手册Molecular CloningA LaboratoryManual.冷泉港实验室出版社，冷泉港，纽约)，利用结合δ-内毒素或δ-内毒素相关蛋白质上存在的一个或多个表位的抗体对转基因植物进行蛋白质印迹和生化检测及类似测定，以证实由δ-内毒素或δ-内毒素相关基因编码的蛋白质的存在。
植物中的杀虫活性在本发明的另一方面，可产生表达具有杀虫活性的δ-内毒素或δ-内毒素相关蛋白质的转基因植物。可利用上述举例说明的方法产生转基因植物，但产生转基因植物细胞的方式对本发明而言不是关键的。可利用诸如农杆菌介导的转化、气溶胶束(aerosol beam)、biolistic转化及非粒子介导方法等本领域中已知或描述的方法来进行，具体所用方法依实验者而定。可用本领域中描述的通用方法分离表达δ-内毒素或δ-内毒素相关蛋白质的植物，例如，通过转化愈伤组织、筛选被转化的愈伤组织以及从所述的转基因愈伤组织再生出可育植物。在所述过程中，可利用任何基因作为可选择标记，只要通过它在植物细胞中的表达能识别或选择已被转化的细胞即可。
已开发了许多与植物细胞一起使用的标记物，诸如对氯霉素、氨基糖苷类G418、潮霉素等等的抗性。编码叶绿体代谢中所涉及的产物的其它基因也可用作选择标记。例如，能提供对诸如草甘膦、溴苯腈(bromoxynil)或咪唑啉酮(imidazolinone)等植物除草剂的抗性的基因可能有具体用途。这样的基因已有报道(Stalker等(1985)J.Biol.Chem.2636310-6314(溴苯腈抗性腈水解酶基因)；以及Sathasivan等(1990)Nucl.Acids Res.182188(AHAS咪唑啉酮抗性基因)。
可对表达δ-内毒素或δ-内毒素相关蛋白质的可育植物测试杀虫活性，并选择显示出最佳活性的植物进行进一步的培育。本领域中有可供利用的方法能够用于检测对害虫的活性。通常，所述蛋白质被混合并用于喂服试验中。参阅，例如，Marrone等(1985)J.ofEconomic Entomology 78290-293。
杀虫控制中的应用将本发明的株系用于杀虫控制或将其它生物体改造成杀虫剂的一般方法是本领域所已知的。参阅，例如，美国专利5,039,523和EP0480762A2。
本发明的芽孢杆菌株系或经遗传改造而含有杀虫基因和蛋白质的微生物可用于保护农作品和产品免受害虫的侵袭。在本发明的一个方面，当将产生毒素的细胞施用于有目标害虫的环境中时，用延长所述细胞中产生的毒素的活性的试剂处理生产毒素(杀虫剂)的生物体的完整(即未裂解)细胞。
或者，通过将异源基因引入细胞宿主而产生杀虫剂。异源基因的表达直接或间接的导致了杀虫剂在细胞内的产生和保持。在本发明的一个方面，当将所述细胞施用于有目标害虫的环境中时，在延长该细胞所产生的毒素的活性的条件下处理这些细胞。所产生的产物保持了毒素的毒性。然后可依照常规技术配制这些自然封装的杀虫剂，以便施用于含有目标害虫的环境中，如，土壤、水和植物的叶子。参阅，例如EPA 0192319以及其中引用的文献。或者，可对表达本发明基因的细胞进行配制，以便可将所产生的物质作为杀虫剂施用。
本发明的活性成分通常以组合物的形式施用，并可与其它化合物同时或接连地施用于待处理的农作物地区或植物。这些化合物可以是肥料、除草剂、防冻剂、表面活性剂、去垢剂、杀虫皂、休眠油、聚合体和/或在单次施用制剂后可长期对目标区域进行给药的随时间释放型或可生物降解型载体制剂。它们也可以是选择性除草剂、化学杀虫剂、杀病毒剂、杀微生物剂、杀变形虫剂、杀虫剂、杀真菌剂、杀菌剂、杀线虫剂、杀软体动物剂或数种这些制剂的混合物，如果需要，可进一步与农业可接受载体、表面活性剂或制备领域中常规采用的促进应用的佐剂一起施用。合适的载体和佐剂可以是固体或液体，并与制备工艺中通常应用的物质相对应，如，天然或再生的矿物、溶剂、分散剂、增湿剂、粘附剂、粘合剂或肥料。同样的，所述制剂可制备成可食用的“饵”或作成害虫“陷阱”，使得可被杀虫制剂的目标害虫所饲用或摄取。
应用本发明活性成分或含本发明细菌菌株所产生的杀虫性蛋白质中至少一种的本发明农用化学品组合物的优选方法是叶片施用、种子包裹和土壤施用。施用次数和施用频率取决于相应害虫的侵袭强度。
所述组合物可制备成粉剂、尘剂、丸剂、颗粒剂、喷雾剂、乳剂、胶体、溶液等等，且可用诸如含所述多肽的细胞培养物的干燥、冻干、匀浆、提取、过滤、离心、沉降和浓缩等常规方式制备。在所有含至少一种所述杀虫多肽的所述组合物中，所述多肽可以重量约1％-约99％的浓度存在。
利用本发明方法可杀死鳞翅类或鞘翅类害虫，或者减少其在特定区域内的数目，或者可预防性的施用于周围区域以防止易感害虫的蔓延。优选的，所述害虫摄取或接触杀虫有效量的所述多肽。“杀虫有效量”指能导致至少一种害虫死亡或能显著降低害虫的生长、饲用或正常生理发育的杀虫剂用量。这个量将取决于各种因素而变化，诸如待控制的具体目标害虫；待处理的特异环境、位置、植物、农作物或农业场所、环境条件，以及该杀虫有效性多肽组合物施用的方法、速率、浓度、稳定性和用量。制剂还可随气候条件、环境因素和/或施用频率和/或害虫侵袭的严重程度等而发生变化。
所述的杀虫剂组合物可通过制备细菌细胞、晶体和/或孢子悬浮液或者与所需的农业可接受载体有组合的分离蛋白质组分制成。所述组合物可在施用前配制成适当的形式，诸如冻干、冷冻干燥、脱水或在水性载体、培养基或诸如盐水或其它缓冲液等适当的溶剂中。制备好的组合物可以是尘土或颗粒物形式，或者油(植物油或矿物油)中的悬浮液，或者水或油/水乳剂，或作为可湿粉末，或者与适于农业应用的任何其它载体物质相组合。适当的农业载体可以是固体或液体，在本领域中是众所周知的。术语“农业可接受载体”涵盖了通常应用于杀虫剂制备工艺中的所有佐剂、惰性成分、分散剂、表面活性剂、粘着剂、粘合剂等，这些是杀虫剂制备业中的技术人员众所周知的。所述制剂可与一种或多种固体或液体佐剂混合，并通过各种方式制备，例如，用常规的制备工艺将杀虫组合物与适当的佐剂均匀混合、混和和/或碾磨。合适的制剂和应用方法参阅美国专利6,468,523，该文件在此收编作为参考。
“害虫”包括但不局限于，昆虫、真菌、细菌、线虫、螨、蜱等等。昆虫类的害虫包括选自以下目的昆虫鞘翅目、双翅目、膜翅目、鳞翅目、食毛目、同翅目、半翅目、直翅目、缨翅目、革翅目、等翅目、虱目、蚤目、毛翅目等，尤其是鞘翅目、鳞翅目和双翅目。
昆虫类害虫包括选自以下目的昆虫鞘翅目、双翅目、膜翅目、鳞翅目、食毛目、同翅目、半翅目、直翅目、缨翅目、革翅目、等翅目、虱目、蚤目、毛翅目等，尤其是鞘翅目和鳞翅目。针对主要农作物的本发明昆虫类害虫包括玉米Ostrinia nubilalis，欧洲玉米螟；Agrotis ipsilon，小地虎；Helicoverpa zea，谷实夜蛾；Spodopterafrugiperda，草地夜蛾；Diatraea grandiosella，巨座玉米螟(西南玉米杆草螟)；Elasmopalpus lignosellus，南美玉米苗斑螟(小玉米螟)；Diatraea saccharalis，小蔗杆草螟；Diabroticavirgifera，玉米(幼芽)根叶甲；Diabrotica longicornis barberi，长角叶甲；Diabrotica undecimpunctata howardi，黄瓜十一星叶甲；Melanotus spp.，梳爪叩头虫；Cyclocephala borealis，北方圆头犀金龟(圆头独角仙)；Cyclocephala immaculata，南方圆头犀金龟(圆头无斑独角仙)；Popillia japonica，弧丽蛂(日本丽金龟)；Chaetocnema pulicaria，玉米(铜色)跳甲；Sphenophorus maidis，玉米隐喙象(玉米谷象)；Rhopalosiphum maidis，玉米蚜(玉米缢管蚜)；Anuraphis maidiradicis，玉米根蚜；Blissus leucopterusleucopterus，麦长蝽(美洲谷长蝽)；Melanoplus femurrubrum，赤胫黑蝗(赤腿蚱蜢)；Melanoplus sanguinipes，血黑蝗(迁徙蚱蜢)；Hylemya platura，(玉米)种蝇；Agromyza parvicornis，玉米斑潜蝇；Anaphothrips obscrurus，玉米黄呆蓟马(美洲锥尾蓟马)；Solenopsis milesta，窃蚁；Tetranychus urticae，二斑叶螨(普通红叶螨)；高粱Chilo partellus，玉米禾螟；Spodopterafrugiperda，草地夜蛾；Helicoverpa zea，谷实夜蛾；Elasmopalpuslignosellus，南美玉米苗斑螟(小玉米螟)；Feltia subterranea，粒肤地虎；Phyllophaga crinita，蛴螬(金龟幼虫)；Eleodes，宽胸叩甲(Conoderus)和Aeolus spp.，金针虫(叩甲幼虫)；Oulemamelanopus，黑角负泥虫(橙足负泥虫)；Chaetocnema pulicaria，玉米(铜色)跳甲；Sphenophorus maidis，玉米隐喙象(玉米谷象)；Rhopalosiphum maidis，玉米蚜(玉米缢管蚜)；Sipha flava，美甘蔗伪毛蚜；Blissus leucopterus leucopterus，麦长蝽(美洲谷长蝽)；Contarinia sorghicola，高粱瘿蚊；Tetranychuscinnabarinus，红叶螨(朱砂叶螨)；Tetranychus urticae，叶螨(普通红叶螨)；小麦Pseudaletia unipunctata，粘虫；Spodoptera frugiperda，草地夜蛾；Elasmopalpus lignosellus，南美玉米苗斑螟(小玉米螟)；Agrotis orthogonia，西方灰地老虎；Elasmopalpus lignosellus，南美玉米苗斑螟；Oulema melanopus，黑角负泥虫(橙足负泥虫)；Hypera punctata，三叶草叶象(车轴草叶象)；Diabrotica undecimpunctata howardi，黄瓜十一星叶甲；俄国麦长管蚜；Schizaphis graminum，麦二岔蚜；Macrosiphumavenae，麦长管蚜；Melanoplus femurrubrum，赤胫黑蝗(赤腿蚱蜢)；Melanoplus differentialis，异黑蝗(殊种蚱蜢)；Melanoplussanguinipes，血黑蝗(迁徙蚱蜢)；Mayetiola destructor，小麦瘿蚊；Sitodiplosis mosellana，麦红吸浆虫；Meromyza americana，美洲杆蝇(美洲麦杆黄潜蝇)；Hylemya coarctata，麦瘿种蝇(冬作种蝇)；Frankliniella fusca，烟褐花蓟马(烟草褐蓟马)；Cephuscinctus，麦茎蜂；Aceria tulipae，麦瘿螨(郁金香瘤螨)；向日葵Suleima helianthana，向日葵芽蛀虫；Homoeosoma electellum，向日葵(同)斑螟；zygogramma exclamationis，向日葵叶甲；Bothyrus gibbosus，胡萝卜利犀金龟(胡萝卜金龟)；Neolasiopteramurtfeldtiana，向日葵籽瘿蚊；棉花Heliothis virescens，烟芽夜蛾；Helicoverpa zea，棉铃虫；Spodoptera exigua，甜菜夜蛾；Pectinophora gossypiella，红铃麦蛾(棉红铃虫)；Anthonomusgrandis，墨西哥棉铃象(甲)；Aphis gossypii，棉蚜；Pseudatomoscelis seriatus，棉序盲蝽(棉跳盲蝽)；Trialeurodesabutilonea，苘麻粉虱(结翅粉虱)；Lygus lineolaris，(美国)牧草盲蝽；Melanoplus femurrubrum，赤胫黑蝗(赤腿蚱蜢)；Melanoplus differentialis，异黑蝗(殊种蚱蜢)；Thrips tabaci，烟蓟马(葱蓟马)；Franklinkiella fusca，烟褐花蓟马(烟草褐蓟马)；Tetranychus cinnabarinus，红叶螨(朱砂叶螨)；Tetranychusurticae，二斑叶螨(普通红叶螨)；稻Diatraea saccharalis，小蔗杆草螟；Spodoptera frugiperda，草地夜蛾；Helicoverpa zea，棉铃虫；Colaspis brunnea，葡萄肖叶甲；Lissorhoptrus oryzophilus，稻根象；Sitophilus oryzae，米象；Nephotettix nigropictus，黑尾叶蝉；Blissus leucopterus leucopterus，麦长蝽(美洲谷长蝽)；Acrosternum hilare，拟缘椿(喜绿蝽)；大豆Pseudoplusiaincludens，大豆(尺)夜蛾；Anticarsia gemmatalis，梨豆夜蛾；Plathypena scabra，苜蓿绿夜蛾；Ostrinia nubilalis，欧洲玉米螟；Agrotis ipsilon，小地虎；Spodoptera exigua，甜菜夜蛾；Heliothis virescens，烟芽夜蛾；Helicoverpa zea，棉铃虫；Epilachna varivestis，墨西哥(大)豆瓢虫；Myzus persicae，桃蚜；Empoasca fabae，蚕豆微叶蝉；Acrosternum hilare，拟缘蝽(喜绿蝽)；Melanoplus femurrubrum，赤胫黑蝗(赤腿蚱蜢)；Melanoplusdifferentialis，异黑蝗(殊种蚱蜢)；Hylemya platura，(玉米)种蝇；Sericothrips variabilis，大豆牧草虫；Thrips tabaci，烟蓟马(葱蓟马)；Tetranychus turkestani，土耳其叶螨(草莓叶螨)；Tetranychus urticae，二斑叶螨(普通红叶螨)；大麦Ostrinianubilalis，欧洲玉米螟；Agrotis ipsilon，小地虎；Schizaphisgraminum，麦二岔蚜；Blissus leucopterus leucopterus，麦长蝽(美洲谷长蝽)；Acrosternum hilare，拟缘椿(喜绿蝽)；Euschistusservus，烟草蝽(褐臭蝽)；Delia platura，(灰地)种蝇；Mayetioladestructor，小麦瘿蚊；Petrobia latens，麦岩螨；油籽油菜Brevicoryne brassicae，甘蓝蚜(菜蚜)；Phyllotreta cruciferae，十字花跳菜甲；Mamestra configurata，蓓带夜蛾(披肩夜蛾)；Plutella xylostella，菜蛾；Delia ssp.，地种蝇(花蝇)。
线虫类包括寄生性线虫，例如根节、胚囊和损害线虫，包括异皮线虫(Heterodera spp.)、根结线虫(Meloidogyne spp.)和球异皮线虫(Globodera spp.)，具体而言是胚囊线虫类，包括但不局限于，Heterodera glycines(大豆异皮线虫)、Heterodera schachtii(甜菜胞囊线虫)、Heterodera avenae(燕麦胞囊线虫)和Globoderarostochiensis(马铃薯金线虫)以及苍白球异皮线虫(Globoderapailida)。损害线虫包括短杆线虫(Pratylenchus spp.)。
以下实施例仅供例证说明而非起限制作用。
实施例实施例1质粒DNA的抽提以如下方式由菌株ATX 13026或ATX 13002制备质粒DNA。在大量的丰富培养基中培养菌株ATX 13026或ATX 13002的纯培养物。将培养液离心以收获细胞沉淀块。然后以本领域已知方法用SDS处理细胞沉淀块，导致细胞壁破损且DNA释放。然后通过高盐浓度沉淀蛋白质和大基因组DNA。通过标准乙醇沉淀来沉淀质粒DNA。通过在氯化铯梯度中的高速离心将质粒DNA与任何剩余的染色体DNA分开。使用UV光显现梯度中的DNA，并使用注射器吸取较低密度条带(即较低条带)。此条带含有来自菌株(或是ATX 13026或是ATX 13002)的质粒DNA。利用本领域已知方法通过在琼脂糖凝胶上的显影来检查DNA的质量。
实施例2基因的克隆正如本领域已知的，将纯化的质粒DNA剪切成5-10kb大小的片段，并且在存在所有四种dNTP的条件下使用T4DNA聚合酶和Klenow片段修补5′和3′单链突出。然后，如本领域已知的，通过T4多核苷酸激酶处理而在5′末端添加磷酸根。如本领域已知的，将修复的DNA片段过夜连接到适当制备以接受插入片段(例如通过限制酶消化而生成平端)的标准高拷贝载体(即pBluescript SK+)中。
通过用已知在克隆位点侧翼具有切割位点的限制酶消化克隆亚群来分析文库的质量。确定了高百分比的克隆含有插入片段，平均长度5-6kb。
实施例3文库平板的高通量测序将通过上述方法制备的文库涂布在含有适当抗生素的丰富培养基上以维持质粒克隆，并将菌落单独挑到装有2ml含有适当抗生素的培养基的96孔板中。将这些板于37℃、以350rpm摇速培养过夜。将板离心而将细胞收集到板底。通过标准的碱裂解法以高通量方式由这些培养物分离质粒DNA。
以如下方式对来自每块板的大量克隆测定来自该文库的克隆的末端序列。使用荧光染料终止物测序技术(Applied Biosystems)和位于克隆位点每一侧侧翼的标准引物测定选择用于分析的每个克隆的DNA序列。一旦反应在热循环仪中完成，则使用标准乙醇沉淀法来沉淀DNA。将DNA重悬于水，并加载到毛细管测序仪上。由克隆位点的任一侧测定每个文库平板的DNA，每个平板的每个插入片段产生两个阅读结果。
实施例4测序数据的装配和筛选将得到的DNA序列汇编成装配方案，并排列在一起以形成毗连序列群。这可以使用电脑程序有效进行，诸如Vector NTi，或者使用DNA比对和分析程序的Pred/Phrap套餐。将这些毗连序列群连同未能添加到毗连序列群中的任何个别阅读序列与所有类型的已知杀虫剂基因的汇编数据库进行比较。进一步分析鉴定为与已知内毒素或杀虫剂基因具有同一性的毗连序列群或个别阅读序列。在得到的序列中，克隆pAX004、pAX006、pAX007、pAX008、pAX009和pAX014含有鉴定为与已知内毒素基因具有同源性的DNA。因此，选择这些克隆用于进一步测序。
实施例5δ-内毒素基因的测序和鉴定设计与内毒素基因具有同源性的序列退火的引物，使得由这些引物产生的DNA序列将与克隆中的现有DNA序列交叠。这一称为“寡聚物行走”的方法在本领域是众所周知的。该方法被用于测定显示与已知内毒素基因有同源性的区域的完整DNA序列。在上文所述克隆的情况中，该方法被用于测定整个克隆的DNA序列，从而得到每个基因的单一核苷酸序列。然后将完整DNA序列放回最初的大装配结果中以进一步确认。这能够将更多的DNA序列阅读结果插入毗连序列群，产生覆盖整个区域的多个阅读结果。
对与已知内毒素基因同源的每个区域进行的DNA序列分析鉴定得到了与已知δ-内毒素基因同源的开放读码框。将由pAX004鉴定得到的开放读码框命名为AXMI-004。将由pAX006鉴定得到的开放读码框命名为AXMI-006。将由pAX007鉴定得到的开放读码框命名为AXMI-007。将由pAX008鉴定得到的开放读码框命名为AXMI-008。将由pAX009鉴定得到的开放读码框命名为AXMI-009。将由pAX014鉴定得到的开放读码框命名为AXMI-014。SEQ ID NOS1和2提供了AXMI-004的DNA序列，而SEQ ID NO3提供了预测的AMXI-004蛋白质的氨基酸序列。位于SEQ ID NO1第385位核苷酸处的AXMI-004备选起始位点产生了SEQ ID NO5中提供的氨基酸序列。SEQ ID NOS6提供了AXMI-006的DNA序列，而SEQ ID NO7提供了预测的AMXI-006蛋白质的氨基酸序列。SEQ ID NO8提供了AXMI-007的DNA序列，而SEQ ID NO9提供了预测的AMXI-007蛋白质的氨基酸序列。位于SEQ IDNO8第151位核苷酸处的AXMI-007备选起始位点产生了SEQ ID NO11中提供的氨基酸序列。SEQ ID NOS12和13提供了AXMI-008的DNA序列，而SEQ ID NO14提供了预测的AMXI-008蛋白质的氨基酸序列。位于SEQ ID NO12第177位核苷酸处的AXMI-008备选起始位点产生了SEQ ID NO16提供中的氨基酸序列。进一步的分析鉴定得到了紧临AXMI-008开放读码框3′末端的开放读码框。SEQ ID NO18中提供了在本文中称为AXMI-008orf2的该开放读码框的这一预测的氨基酸序列。SEQ ID NOS19中提供了AXMI-009的DNA序列，而SEQ ID NO20中提供了预测的AMXI-009蛋白质的氨基酸序列。位于SEQ ID NO19第34位核苷酸处的AXMI-009备选起始位点产生了SEQ ID NO22中提供的氨基酸序列。位于SEQ ID NO19第64位核苷酸处的另一个AXMI-009备选起始位点产生了SEQ ID NO24中提供的氨基酸序列。SEQ ID NOS25和26中提供了AXMI-014的DNA序列，而SEQ ID NO27提供了预测的AMXI-008蛋白质的氨基酸序列。位于SEQ ID NO25第136位核苷酸处的AXMI-014备选起始位点产生了SEQ ID NO29中提供的氨基酸序列。
实施例6分离基因与已知内毒素基因的同源性用本发明的DNA序列和氨基酸序列搜索DNA和蛋白质数据库揭示出了这些序列与已知内毒素同源。
AXMI-004图1显示了AXMI-004与几种内毒素的序列比对。Blast搜索确定了cry1Ca具有最强的同源区，在毒性结构域的整体序列同一性为43％(见表1)。
表1.AXMI-004与示范性内毒素类型的氨基酸同一性
AXMI-006图2显示了AXMI-006与几种内毒素的序列比对。Blast搜索确定了cry4Aa具有最强的同源区，尽管AMXI-006蛋白质(SEQ ID NO7)与一大组内毒素蛋白质的序列比对显示最同源的蛋白质是cry10Aa。cry10Aa与AXMI-006的整体氨基酸同一性是25％(见表2)。对AXMI-006氨基酸序列的检查提示它不含几种内毒素家族中存在的C-末端非毒性结构域。由序列比对中除去毒素的这个C-末端蛋白质后，序列比对反映了氨基酸同一性只存在于毒素结构域中(见表2，第3列)。图2显示了这一“修剪过的”序列比对。
表2.AXMI-006与示范性内毒素类型的氨基酸同一性
AXMI-007
Blast搜索确定了cry4Aa具有与AXMI-007最强的同源区，尽管AMXI-007蛋白质(SEQ ID NO9)与一大组内毒素蛋白质的序列比对显示最同源的蛋白质是cry10Aa。cry10Aa与AXMI-007的整体氨基酸同一性是25％(见表3)。对AXMI-007氨基酸序列的检查提示它不含几种内毒素家族中存在的C-末端非毒性结构域。由序列比对中除去毒素的这个C-末端蛋白质后，序列比对反映了氨基酸同一性只存在于毒素结构域中(见表3，第3列)。图3显示了这一“修剪过的”序列比对。
表3.AXMI-007与示范性内毒素类型的氨基酸同一性
AXMI-008Blast搜索确定了cry40Aa具有与AXMI-008最强的同源区，而且AMXI-008蛋白质(SEQ ID NO14)与一大组内毒素蛋白质的序列比对显示最同源的蛋白质是cry40Aa。cry40Aa与AXMI-008的整体氨基酸同一性是66％(见表4)。对AXMI-008氨基酸序列的检查提示它不含几种内毒素家族中存在的C-末端非毒性结构域。由序列比对中除去毒素的这个C-末端蛋白质后，序列比对反映了氨基酸同一性只存在于毒素结构域中(见表4，第3列)。图4显示了这一“修剪过的”序列比对。
表4.AXMI-008与示范性内毒素类型的氨基酸同一性
紧临AXMI-008编码区下游(3′)的开放读码框与已知内毒素相关蛋白质具有同源性。Blast搜索确定了crybun3orf2(cry40Aa的下游开放读码框)具有最强的同源区。数据库中存在几种其它orf-2样蛋白质，而且图5显示了AMXI-008-orf2蛋白质(SEQ ID NO18)与这些蛋白质中的一组的序列比对。这些蛋白质还与cry4Aa和cry4Ba C-末端非毒性结构域享有同源性。AXMI-8-orf2与cry40Aaorf2的整体氨基酸同一性是86％(见表5)。
表5.AXMI-008-orf2与相关蛋白质的氨基酸同一性
AXMI-009Blast搜索确定了cryBAa具有与AXMI-009最强的同源区，而且AMXI-009蛋白质(SEQ ID NO20)与一大组内毒素蛋白质的序列比对显示最同源的蛋白质是cry3Ba和cry16Aa。cry3Ba和cry16Aa与AXMI-009的整体氨基酸同一性是26％(见表6)。对AXMI-009氨基酸序列的检查提示它不含几种内毒素家族中存在的C-末端非毒性结构域。由序列比对中除去毒素的这个C-末端蛋白质后，序列比对反映了氨基酸同一性只存在于毒素结构域中(见表6，第3列)。图6显示了这一“修剪过的”序列比对。
表6.AXMI-009与示范性内毒素类型的氨基酸同一性
AXMI-014Blast搜索确定了cry40Aa具有与AXMI-014最强的同源区，而且AMXI-0014蛋白质(SEQ ID NO27)与一大组内毒素蛋白质的序列比对显示最同源的蛋白质是cry40Aa。cry40Aa与AXMI-014的整体氨基酸同一性是55％(见表7)。对AXMI-014氨基酸序列的检查提示它不含几种内毒素家族中存在的C-末端非毒性结构域。由序列比对中除去毒素的这个C-末端蛋白质后，序列比对反映了氨基酸同一性只存在于毒素结构域中(见表7，第3列)。图7显示了这一“修剪过的”序列比对。
表7.AXMI-014与示范性内毒素类型的氨基酸同一性
实施例7AXMI-006与AXMI-007之间的同源性对氨基酸序列AXMI-007与AXMI-006的比较显示了这两种毒素共享显著的氨基酸同源性。氨基酸序列AXMI-006(SEQ ID NO7)和AXMI-007(SEQ ID NO9)的比对显示了这两种蛋白质在氨基酸水平上85％相同。因此，AXMI-006和AXMI-007组成一类新的相关内毒素。
实施例8杀虫活性的测定法常常以多种方式评估杀虫剂蛋白质作为杀虫剂作用于害虫的能力。本领域众所周知的一种方法是进行喂食测定法。在这种喂食测定法中，将害虫暴露于含有待测化合物的样品或对照样品。通常，这是通过将待测物质或其合适稀释物置于害虫将要摄食的物质诸如人工饲料上来进行的。待测物质可以由液体、固体或浆液构成。可以将待测物质置于表面之上，然后使其干燥。或者，可以将待测物质与熔化的人工饲料混和，然后分装到测定槽中。测定槽可以是例如杯、盘或微量滴定板的孔。
针对吸食害虫(例如蚜虫)的测定法可以包括使用隔离物将测试物质与昆虫分开，理想的是吸食昆虫的吸食口器能够刺穿的物体，从而能够摄食测试物质。通常，将测试物质与喂食刺激物诸如蔗糖混和以促进测试化合物的摄食。
其它类型的测定法可以包括将测试物质显微注射到害虫的口或肠中，以及开发转基因植物，随后测试害虫以转基因植物为食的能力。植物测试可以包括分离通常取食的植物部分，例如将小笼附于叶片上，或者在装有昆虫的笼中分离整个植株。
本领域还知道测定害虫的其它方法和途径，可以在如下文献中找到Robertson，J.L.和H.K.Preisler，1992，《使用节肢动物的杀虫剂生物测定法》(Pesticide bioassays with arthropods)，CRC，伯克莱屯，佛罗里达州。或者，测定法一般描述于如下期刊中“Arthropod Management Tests”和“Journal of EconomicEntomology”，或者是与美国昆虫学学会(Entomological Society ofAmerica，ESA)成员的讨论。
实施例9克隆AXMI-006用于蛋白质表达如下将AXMI-006克隆到用于大肠杆菌表达的载体中。pAX480含有用于选择转化子的卡那霉素抗性基因，以及可通过IPTG诱导来调控蛋白质表达的tac启动子。通过将编码6xHis标签区的DNA片段紧临着插入插入片段克隆区的上游来修饰pAX480，使得得到的克隆在所表达的蛋白质的N端含有6xHis标签。本领域众所周知表达含有6xHis标签融合体的蛋白质的方法及其用于纯化和分析蛋白表达的用途。
使用PfuUltraTM高保真DNA聚合酶(Stratagene)PCR扩增AXMI-006的编码序列。设计寡核苷酸引物，使得得到的PCR产物在两个末端附近含有期望的限制性位点，以便于克隆。用合适的限制酶消化所得到的PCR产物(约2.2kb)，并亚克隆到经过修饰的pAX480中。通过限制性分析鉴定含有插入片段的克隆。得到的克隆pAX906含有与6xHis标签融合的AXMI-006开放读码框，其转录和翻译导致生成具有一段六个组氨酸的“融合蛋白”。通过DNA序列分析确认了pAX906的DNA序列，随后如制造商(Stratagene，拉霍亚，加利福尼亚州)所述转化到化学感受态大肠杆菌BL21中。
将pAX906在BL21中的单菌落接种到添加了卡那霉素的LB培养基中，并于37℃以剧烈振摇培养几个小时。将这些培养物培养至OD600为0.6-0.8，然后通过添加0.1mM IPTG来诱导蛋白质生成。将培养物在诱导条件下培养3小时，然后通过离心沉淀细胞并重悬于PBS。采用10秒超声间隔，使用Misonix Sonicator 3000超声处理细胞，总计30秒，并在冰上保温1分钟。
实施例10AXMI-006对烟芽夜蛾和草地夜蛾的活性的生物测定法使用人工饲料(多物种饲料，Southland Products，Lake Village，阿肯色州)进行经过超声处理的pAX906培养物的生物测定法。通过将经过超声处理的pAX906细胞或作为对照的大肠杆菌菌株的细胞施加到饲料表面并使饲料表面干燥来进行生物测定法。在24孔组织培养板中进行生物测定法。生物测定法在黑暗中、25℃和65％相对湿度下进行。用Breathe简易密封带(Diversified Biotech，波士顿，马萨诸塞州)密封盘子。第5天记录结果。
表8.AXMI-006对草地夜蛾的生物测定法
表9.AXMI-006对烟芽夜蛾的生物测定法
阻碍发育被定义为昆虫体形变小，而且一些幼虫摄食减少。发育受阻碍的昆虫还可能表现出相对于未处理饲料而言回避经过处理的饲料。
实施例11AXMI-008、AXMI-009和AXMI-014对粉纹夜蛾的杀虫活性将含有pAX008、pAX009或pAX-014的大肠杆菌菌株，以及未转化的大肠杆菌的培养物在2ml LB培养基(Luria-Bertani肉汤，BectonDickinson & Company，斯帕克斯，马里兰州)中于37℃以250rpm振摇培养24小时。将含有质粒的菌株在含有适当抗生素的LB培养基中培养，以选择质粒在大肠杆菌中的维持。
使用人工饲料(多物种饲料，Southland Products，Lake Village，阿肯色州)在24孔组织培养板中进行生物测定法。通过将含有pAX-014的大肠杆菌培养物施加到饲料表面并使饲料表面干燥来进行生物测定法。将菌株的整个培养物以每孔40μl培养物的浓度施加于饲料。生物测定法在黑暗中、25℃和65％相对湿度下进行。用Breathe简易密封带(Diversified Biotech，波士顿，马萨诸塞州)密封盘子。第5天记录结果。
表10.对粉纹夜蛾的杀虫活性
实施例12δ-内毒素基因在芽孢杆菌中的表达通过PCR由来自实施例4的克隆扩增AXMI-004、AXMI-006、AXMI-007、AXMI-008和AXMI-009，并通过本领域众所周知的方法克隆到芽孢杆菌表达载体pAX916中。对于AXMI-004，将得到的克隆命名为pAX920。对于AXMI-006，将得到的克隆命名为pAX921。对于AXMI-007，将得到的克隆命名为pAX919。对于AXMI-008，将得到的克隆命名为pAX922。对于AXMI-009，将得到的克隆命名为pAX917。所得到的克隆在转化到cry(-)苏云金芽孢杆菌菌株的细胞中后表达相应蛋白质。可以在多种常规培养基中培养含有δ-内毒素基因且表达δ-内毒素蛋白质的芽孢杆菌菌株。在CYS培养基(10g/lBacto-casitone；3g/l酵母提取物；6g/l KH2PO4；14g/l K2HPO4；0.5mM MgSO4；0.05mM MnCl2；O.05mM FeSO4)中培养含有期望基因的芽孢杆菌菌株，直至通过显微镜检查观察到孢子形成。制备样品，并在针对重要昆虫害虫的生物测定法中对δ-内毒素蛋白质测试杀虫活性。
方法配制CYS培养基10g/l Bacto-casitone；3g/l酵母提取物；6g/l KH2PO4；14g/l K2HPO4；0.5mM MgSO4；0.05mM MnCl2；0.05mMFeSO4。CYS混合物应当是pH7，如果需要就进行调节。优选用NaOH或HCl。然后将培养基高压灭菌，并在高压灭菌后添加100ml 10X经过滤的葡萄糖。如果得到的溶液浑浊，可以在室温搅动至清澈。
实施例13在芽孢杆菌中表达的AXMI-004的N端氨基酸序列对于在芽孢杆菌中表达的AXMI-004的分析说明在这些培养物中检测到的蛋白质产物在长度上相对于全长AXMI-004蛋白质而言可能是缩短的。因为许多内毒素蛋白质在芽孢杆菌中表达后在N端受到切割，我们测定了由芽孢杆菌表达产生的AXMI-004蛋白质的N末端。在PAGE凝胶上分离来自AXMI-004的蛋白质样品，并且通过本领域众所周知的方法将蛋白质转移到PVDF膜上。切下与AXMI-004对应的蛋白质条带。通过本领域已知的连续Edman降解法测定该蛋白质的N端氨基酸序列。得到的序列如下ERFDKNDALE该氨基酸序列与全长AXMI-004序列(SEQ ID NO3)的比较证明该氨基酸序列来自AXMI-004在芽孢杆菌中表达后的内在切割，导致N末端与SEQ ID NO3第28位氨基酸相对应的蛋白质(公开于SEQ IDNO5)。
实施例14AXMI-004对昆虫害虫的生物测定法利用公认的生物测定法流程，使用形成孢子的表达AXMI-004的芽孢杆菌细胞培养物裂解物确定AXMI-004的杀虫活性。在50ml CYS培养基中培养芽孢杆菌培养物，以用于标准生物测定法和LC50生物测定法。然后将培养物于30℃、以250rpm培养2-3天，直至细胞形成孢子。通过显微镜检查孢子的存在来确定孢子形成。通过将形成孢子的培养物以12,000xg离心10分钟来制备AXMI-004蛋白质样品。收集沉淀并重悬于4ml 20mM Tris-HCl，pH8.0。将管置于冰上的同时将悬浮液超声处理20秒(大功率，使用微型探针)。通过在SDS 4-20％梯度丙烯酰胺凝胶上与已知量的牛血清清蛋白(BSA)一起电泳来测定样品的蛋白质浓度(图8)。AXMI-004的浓度经测定是0.4μg/μl。
使用按照本领域所知制备的人工饲料(多物种饲料，SouthlandProducts，Lake Village，阿肯色州)的表面处理生物测定法来测试AXMI-004的杀虫活性。通过将表达AXMI-004的芽孢杆菌培养物施加到饲料表面并使表面通风干燥来进行生物测定法。在标准生物测定法中，每个孔使用5个虫卵，而LC50生物测定法每个孔使用10条新生昆虫幼虫。使用细尖毛刷施加虫卵或幼虫。标准表面生物测定法在24孔组织培养板中进行。将40ul每种样品施加到每个孔中。因为每个孔的表面积是2cm2(板的来源)，40μl细胞裂解物样品含有大约0.4ug/ul AXMI-004。测定LC50的生物测定法在48孔组织培养板中进行，每个孔展示的表面积是1cm2(来源)，每个孔使用大约20ul 0.4μg/ulAXMI-004。每个生物测定法中AXMI-004蛋白质的最终量是大约8μg/cm2。用Breathe简易密封带(Diversified Biotech，波士顿，马萨诸塞州)密封生物测定法的板。对照样品包括只含培养基的样品，以及未经样品处理的孔。然后将生物测定法在黑暗中于25℃和65％相对湿度下放置5天，并记录结果。
表11.AXMI-004的杀虫活性
AXMI-004显示针对欧洲玉米螟和烟芽夜蛾有强杀虫活性(100％致死)。AXMI-004还显示针对红铃麦蛾有50-75％致死的杀虫活性。浓度为43μg/cm2的AXMI-004对红铃麦蛾产生70％死亡率。AXMI-004严重阻碍小地虎、棉铃虫和草地夜蛾的发育。
实施例15AXMI-004针对烟芽夜蛾和欧洲玉米螟的杀虫活性的量化通过在昆虫生物测定法中测试一定范围的AXMI-004蛋白质浓度并将这些蛋白质样品施加到昆虫饲料表面来测定AXMI-004蛋白质对欧洲玉米螟和烟芽夜蛾幼虫的LC50。记录每个蛋白质浓度的死亡率，并使用Probit分析软件进行分析。结果在95％置信空间是显著的。因为测定是通过表面污染来进行的，在假设全部蛋白质样品在测定过程中都保留在表面上而在昆虫的摄食水平下很少有扩散的前提下测定LC50。因此，测定的数值可能有些低估AXMI-004蛋白质对测试昆虫的毒性。
表12.AXMI-004对欧洲玉米螟的LC50
LC50＝297ng/cm2；95％CI＝218-384.
表13.AXMI-004对烟芽夜蛾的LC50
LC50＝2874ng/cm2；95％CI＝2189-3933实施例16AXMI-004、AXMI-006、AXMI-007和AXMI-009针对牧草盲蝽(Lygus lineolaris)的杀虫活性的量化如下制备细菌裂解物切下芽孢杆菌在50ml CYS培养基中培养60小时。然后将芽孢杆菌培养物以12,000rpm离心10分钟，并丢弃上清液。将沉淀重悬于5ml 20mM Tris HCl pH8。
如下进行生物测定法切下Eppendorf管的底尖和盖子，形成喂食槽。将杀虫剂蛋白质或对照以溶液倒入管帽中并覆盖昆虫能够刺穿从而摄食的石蜡膜(Pechiney Plastic Packaging，芝加哥，伊利诺斯州)从而呈现给昆虫。将Eppendorf管倒置管帽朝下置于黑暗中，并用细尖毛刷将一龄或二龄Lygus nymphs置于Eppendorf槽中。用石蜡膜密封切下的Eppendorf管尖制成测定槽。将得到的测定槽帽端朝下于环境温度保温。在15％葡萄糖溶液中以浓度6.6ug/ml测试杀虫剂蛋白质。
表14.对牧草盲蝽的杀虫活性
实施例17AXMI-008对黄粉虫(Tenebrio molitor)的生物测定法制备表达AXMI-008的芽孢杆菌培养物的样品，并测试对黄粉虫的杀虫活性。当pAX922制备成pH4.0的不溶组分时，它显示对通常称为大黄粉虫的昆虫有活性。
由含有pAX922的芽孢杆菌菌株的培养物制备AXMI-008的样品。如下制备生物测定法样品将培养物在CYS培养基中培养4天，直至孢子形成。将样品以10,000rpm离心10分钟，丢弃上清液。用20mMTris pH8清洗沉淀，并以10,000rpm离心10分钟。丢弃上清液，并将沉淀重悬于3ml 50mM柠檬酸钠和25mM氯化钠及2mM DTT pH4。将样品于37℃保温1小时。保温后，将样品以13,000rpm离心10分钟，丢弃上清液。将沉淀重悬于50mM柠檬酸钠和25mM氯化钠pH4。
使用人工饲料(Southern Corn Rootworm Diet，Bioserv，Frenchtown，新泽西州，#F9757B)在24孔组织培养板中进行样品对黄粉虫的生物测定法。通过表面处理以Axmi008浓度8ug/cm2施加样品，并使之空气干燥。使用细尖毛刷施加昆虫。用Breathe简易密封带(Diversified Biotech，波士顿，马萨诸塞州)密封生物测定盘，并在无光条件下于65％相对湿度和25℃保温7天并记录结果。
表15.AXMI-008对黄粉虫的杀虫活性
*其余黄粉虫发育受到阻碍。发育受阻即观察到幼虫体形较小、生长较慢，而且摄食剧烈减少或仅有最低限度。昆虫还可能表现出相对于未处理饲料而言回避经过处理的饲料。
实施例18AXMI-009蛋白质对鞘翅类害虫的生物测定法通过将40μl不溶组分施加到2cm2饲料表面上、达到最终总蛋白质浓度为8μg/cm2来进行AXMI-009蛋白质制剂的生物测定法。使用南方玉米食虫饲料(Bioserv，Frenchtown，新泽西州，#F9757B)测试玉米根叶甲(Diabrotica virgifera virgifera)和黄瓜十一星叶甲(Diabrotica undecimpunctata)。通过将表达AXMI-009的芽孢杆菌培养物施加到饲料表面并使饲料表面干燥来进行生物测定法。生物测定法在24孔组织培养板中进行。标准生物测定法每个孔使用25个虫卵。将虫卵施加到含有0.1％琼脂和30ug/ml制霉菌素的溶液中。用Breathe简易密封带(Diversified Biotech，波士顿，马萨诸塞州)密封盘子并将盖子盖回盘上。将生物测定法避光于65％相对湿度(RH)和25℃保温7天。对于玉米根叶甲和黄瓜十一星叶甲都看到了不溶组分的活性。对照是培养基、缓冲液1mM Tris pH10.5、和芽孢杆菌表达载体pAX916。
表16.玉米根叶甲
表17.黄瓜十一星叶甲
实施例19用于植物表达的载体将δ-内毒素编码区DNA可操作连接用于在植物中表达的合适启动子和终止子序列。这些序列在本领域是众所周知的，而且可以包括用于在单子叶植物中表达的水稻肌动蛋白启动子或玉米泛素启动子、用于在双子叶植物中表达的拟南芥UBQ3启动子或CaMV 35S启动子、以及nos或PinII终止子。用于构建和鉴定启动子-基因-终止子构建体的技术在本领域同样是众所周知的。
将上文所述植物表达盒与合适的植物选择标记结合使用，以便于选择经转化的细胞和组织，并连接到植物转化载体中。这些载体可以包括用于由农杆菌介导的转化的二元载体或用于气雾剂或biolistic转化的单一质粒载体。
实施例20玉米细胞的转化授粉后8-12天采集玉米穗。由穗分离胚，并将大小为0.8-1.5mm的那些胚用于转化。将胚盾片朝上置于合适的培养基上，诸如DN62A5S培养基(3.98g/L N6盐；1mL/L(1000x储液)N6维生素；800mg/LL-天冬酰胺；100mg/L肌醇；1.4g/L L-脯氨酸；100mg/LCasaminoacids；50g/L蔗糖；1mL/L(1mg/mL储液)2，4-D)，并于25℃在黑暗中保温过夜。
将由此产生的外植体转移到筛网上(每块板30-40粒)，转移到渗透培养基上达30-45分钟，然后转移到光照板上(见例如PCT公开号WO/0138514和美国专利号5,240,842)。
采用基本上如PCT公开号WO/0138514中描述的条件(aerosolbeam accelerator)，使用气溶胶气流加速器，将设计在植物细胞中表达δ-内毒素的DNA构建体加速进入植物组织。光照后，将胚在渗透培养基上保温30分钟，然后置于保温培养基上在黑暗中于25℃保温过夜。为了避免过度损伤经过光照的外植体，在转移至恢复培养基前将它们至少保温24小时。再将胚散布到恢复期培养基上，在黑暗中于25℃保温5天，再转移至选择培养基上。将外植体在选择培养基中保温可长达8周，这取决于所用具体选择方法的本质和特征。选择期后，将由此产生的愈伤组织转移至胚成熟培养基，直至观察到成熟体细胞胚的形成。然后将由此产生的成熟体细胞胚置于低光照下，并通过本领域已知方法启动再生程序。使由此产生的芽在生根培养基上生根，并将由此产生的植株转移至苗圃罐中，繁殖成转基因植物。
材料DN62A5S培养基
用1N KOH/1N KCl将溶液的pH调至pH5.8，添加Gelrite(Sigma)至3g/L，并高压灭菌。冷却至50℃后，添加2ml/L硝酸银(Phytotechnology Labs)的5mg/ml储液。该配方制备大约20块平板。
实施例21通过由农杆菌介导的转化转化到植物细胞中授粉后8-12天采集穗。由穗分离胚，并将大小为0.8-1.5mm的那些胚用于转化。将胚盾片朝上置于合适的培养基上，并在黑暗中于25℃保温过夜。但是，本质上并不必将胚保温过夜。使胚接触含有用于Ti质粒介导的转移的合适载体的农杆菌菌株达5-10分钟，然后铺板在共培养培养基上，在黑暗中于25℃保温3天。共培养后，将外植体转移至恢复期培养基，在黑暗中于25℃保温5天。将外植体在选择培养基中保温可长达8周，这取决于所用具体选择方法的本质和特征。选择期后，将由此产生的愈伤组织转移至胚成熟培养基，直至观察到成熟体细胞胚的形成。然后将由此产生的成熟体细胞胚置于低光照下，并通过本领域已知方法启动再生程序。使由此产生的芽在生根培养基上生根，并将由此产生的植株转移至苗圃罐中，繁殖成转基因植物。
结论正如上文实施例中所示，本发明的δ-内毒素蛋白质针对多种害虫具有活性。AXMI-004对包括欧洲玉米螟、小地虎、棉铃虫、草地夜蛾、烟芽夜蛾、红铃麦蛾、烟草天蛾、粉纹夜蛾、和牧草盲蝽在内的害虫有杀虫活性。AXMI-006对包括烟芽夜蛾、草地夜蛾、和牧草盲蝽在内的害虫有杀虫活性。AXMI-007对包括牧草盲蝽在内的害虫有杀虫活性。AXMI-008对包括黄粉虫和粉纹夜蛾在内的害虫有杀虫活性。AXMI-009对包括牧草盲蝽、粉纹夜蛾、玉米根叶甲、和黄瓜十一星叶甲在内的害虫有杀虫活性。AXMI-014对包括粉纹夜蛾在内的害虫有杀虫活性。
说明书中提到的所有发表物和专利申请指示了本发明所属领域熟练技术人员的技术水平。本文收录所有发表物和专利申请作为参考，其程度与专门和个别指出收录每一项发表物或专利申请作为参考相同。
虽然出于清楚理解的目的通过例示和实施例已经较为详细的描述了上述发明，显然可以在所附权利要求的范围内实践某些改变和修饰。
序列表<110>Carozzi，NadineHargiss，TracyKoziel，Michael G.
Duck，Nicholas B.
Carr，Brian<120>δ-内毒素基因及其使用方法<130>045600/274379<150>60/448,632<151>2003-02-20<150>60/448,633<151>2003-02-20<150>60/448,797<151>2003-02-20<150>60/448,806<151>2003-02-20<150>60/448,810<151>2003-02-20<150>60/448,812<151>2003-02-20<160>52<170>FastSEQ for Windows Version 4.0<210>1<211>2190<212>DNA<213>Bacillus thuringiensis<400>1tatgatgata actttgacaa cagccgcatc cataacgtac tttttcaata ttaattgtat 60ccaagcttat tttgtacaaa ttacacttta ttgtatgcat aatagtttct tattgattga 120gcatataaga aaattttttc atatataagt gaaaatattt ttgcttttct tttttttatt 180taggataaac tttacttgta aacacaagta aatagtttat tagagataga gcctcatttt 240aaaatattta cgaatcctat ctataaaaat ctaattttaa aattattcct tttaaatgta 300atgagtgaat tgaaggggaa atttaagaaa agtactaatc gaacttgttg tttgctaaaa 360ataataaata taggaggaag aggtatgaat tcaaaggaac atgattatct aaaagtttgt 420aatgatttaa gtgacgccaa tattaatatg gaacggtttg ataagaatga tgcactggaa 480attggtatgt ccattgtatc tgaacttatt ggtatgattc caggcggaac agctttgcaa 540tttgtgttta atcaattgtg gtctcgttta ggtgattctg gatggaatgc gttcatggaa 600catgtggagg aattaattga tactaaaata gaagggtatg caaaaaataa agccttatct 660gaattagcag gtatacaaag aaaccttgaa acatatatac aattacgtaa tgaatgggaa 720aatgatattg aaaactcaaa ggctcaaggt aaggtagcta attactatga aagtcttgag 780caggcggttg aaaggagtat gcctcaattt gcagtggaga attttgaagt accactttta 840actgtctatg tgcaagctgc taatcttcat ttattattat taagagatgt ttcagtttat 900ggaaagtgtt ggggatggtc ggagcagaaa attaaaattt attatgataa acagattaag 960tatacccatg aatacacaaa tcattgtgta aattggtata ataaaggact tgagagatta 1020aaaaataaag gttcttctta tcaagattgg tacaattata atcgtttccg tagagaaatg 1080actcttactg ttttagatat cgttgcttta ttcccgcact atgatgtaca aacttatcca 1140ataacaaccg ttgctcagct aacaagggaa gtttatacgg atcctttact taattttaat 1200cctaaattac attctgtgtc tcaattacct agttttagtg acatggaaaa tgcaacaatt 1260agaactccac atctgatgga atttttaaga atgctaacaa tttatacaga ttggtatagt 1320gtgggaagaa actattattg gggaggacat cgcgtgacgt cttaccatgt aggaggagag 1380aatataagat cacctctata tggtagagag gcaaatcaag aggttcctag agatttttat 1440ttttatggac ccgtttttaa gacgttatca aagccgactc taagaccatt acagcagcct 1500
gcaccagctc ctccttttaa tttacgtagc ttagagggag tagaattcca cactcctaca 1560ggtagtttta tgtatcgtga aagaggatcg gtagattctt ttaatgagtt gccgcctttt 1620aatccagttg ggttacctca taaggtatac agtcaccgtt tatgtcatgc aacgtttgtt 1680cgtaaatctg ggacccctta tttaacaaca ggtgccatct tttcttggac acatcgtagt 1740gctgaagaaa ccaatacaat tgaatcaaat attattacgc aaatcccgtt agtaaaagca 1800tatcaaattg ggtcaggcac tactgtaagg aaaggaccag gattcacagg aggggatata 1860cttcgaagaa caggtcctgg aacatttgga gatatgagaa taaatattaa tgcaccatta 1920tctcaaagat atcgtgtaag gattcgttat gcttctacga cagatttaca atttgtcacg 1980agtattaatg ggaccaccat taatattggt aacttcccga aaactattaa taatctaaat 2040actttaggtt ctgagggcta tagaacagta tcgtttagta ctccatttag tttctcaaat 2100gcacaaagca tatttagatt aggtatacaa gcattttctg gagttcaaga agtttatgtg 2160gataaaattg aatttattcc tgttgaatag 2190<210>2<211>1890<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(1890)<400>2atg agt gaa ttg aag ggg aaa ttt aag aaa agt act aat cga act tgt 48Met Ser Glu Leu Lys Gly Lys Phe Lys Lys Ser Thr Asn Arg Thr Cys1 5 10 15tgt ttg cta aaa ata ata aat ata gga gga aga ggt atg aat tca aag 96Cys Leu Leu Lys Ile Ile Asn Ile Gly Gly Arg Gly Met Asn Ser Lys20 25 30gaa cat gat tat cta aaa gtt tgt aat gat tta agt gac gcc aat att 144Glu His Asp Tyr Leu Lys Val Cys Asn Asp Leu Ser Asp Ala Asn Ile35 40 45aat atg gaa cgg ttt gat aag aat gat gca ctg gaa att ggt atg tcc 192Asn Met Glu Arg Phe Asp Lys Asn Asp Ala Leu Glu Ile Gly Met Ser50 55 60att gta tct gaa ctt att ggt atg att cca ggc gga aca gct ttg caa 240Ile Val Ser Glu Leu Ile Gly Met Ile Pro Gly Gly Thr Ala Leu Gln65 70 75 80ttt gtg ttt aat caa ttg tgg tct cgt tta ggt gat tct gga tgg aat 288Phe Val Phe Asn Gln Leu Trp Ser Arg Leu Gly Asp Ser Gly Trp Asn85 90 95gcg ttc atg gaa cat gtg gag gaa tta att gat act aaa ata gaa ggg 336Ala Phe Met Glu His Val Glu Glu Leu Ile Asp Thr Lys Ile Glu Gly100 105 110tat gca aaa aat aaa gcc tta tct gaa tta gca ggt ata caa aga aac 384Tyr Ala Lys Asn Lys Ala Leu Ser Glu Leu Ala Gly Ile Gln Arg Asn115 120 125ctt gaa aca tat ata caa tta cgt aat gaa tgg gaa aat gat att gaa 432Leu Glu Thr Tyr Ile Gln Leu Arg Asn Glu Trp Glu Asn Asp Ile Glu130 135 140aac tca aag gct caa ggt aag gta gct aat tac tat gaa agt ctt gag 480Asn Ser Lys Ala Gln Gly Lys Val Ala Asn Tyr Tyr Glu Ser Leu Glu145 150 155 160cag gcg gtt gaa agg agt atg cct caa ttt gca gtg gag aat ttt gaa 528Gln Ala Val Glu Arg Ser Met Pro Gln Phe Ala Val Glu Asn Phe Glu165 170 175gta cca ctt tta act gtc tat gtg caa gct gct aat ctt cat tta tta 576
Val Pro Leu Leu Thr Val Tyr Val Gln Ala Ala Asn Leu His Leu Leu180 185 190tta tta aga gat gtt tca gtt tat gga aag tgt tgg gga tgg tcg gag 624Leu Leu Arg Asp Val Ser Val Tyr Gly Lys Cys Trp Gly Trp Ser Glu195 200 205cag aaa att aaa att tat tat gat aaa cag att aag tat acc cat gaa 672Gln Lys Ile Lys Ile Tyr Tyr Asp Lys Gln Ile Lys Tyr Thr His Glu210 215 220tac aca aat cat tgt gta aat tgg tat aat aaa gga ctt gag aga tta 720Tyr Thr Asn His Cys Val Asn Trp Tyr Asn Lys Gly Leu Glu Arg Leu225 230 235 240aaa aat aaa ggt tct tct tat caa gat tgg tac aat tat aat cgt ttc 768Lys Asn Lys Gly Ser Ser Tyr Gln Asp Trp Tyr Asn Tyr Asn Arg Phe245 250 255cgt aga gaa atg act ctt act gtt tta gat atc gtt gct tta ttc ccg 816Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Val Ala Leu Phe Pro260 265 270cac tat gat gta caa act tat cca ata aca acc gtt gct cag cta aca 864His Tyr Asp Val Gln Thr Tyr Pro Ile Thr Thr Val Ala Gln Leu Thr275 280 285agg gaa gtt tat acg gat cct tta ctt aat ttt aat cct aaa tta cat 912Arg Glu Val Tyr Thr Asp Pro Leu Leu Asn Phe Asn Pro Lys Leu His290 295 300tct gtg tct caa tta cct agt ttt agt gac atg gaa aat gca aca att 960Ser Val Ser Gln Leu Pro Ser Phe Ser Asp Met Glu Asn Ala Thr Ile305 310 315 320aga act cca cat ctg atg gaa ttt tta aga atg cta aca att tat aca 1008Arg Thr Pro His Leu Met Glu Phe Leu Arg Met Leu Thr Ile Tyr Thr325 330 335gat tgg tat agt gtg gga aga aac tat tat tgg gga gga cat cgc gtg 1056Asp Trp Tyr Ser Val Gly Arg Asn Tyr Tyr Trp Gly Gly His Arg Val340 345 350acg tct tac cat gta gga gga gag aat ata aga tca cct cta tat ggt 1104Thr Ser Tyr His Val Gly Gly Glu Asn Ile Arg Ser Pro Leu Tyr Gly355 360 365aga gag gca aat caa gag gtt cct aga gat ttt tat ttt tat gga ccc 1152Arg Glu Ala Asn Gln Glu Val Pro Arg Asp Phe Tyr Phe Tyr Gly Pro370 375 380gtt ttt aag acg tta tca aag ccg act cta aga cca tta cag cag cct 1200Val Phe Lys Thr Leu Ser Lys Pro Thr Leu Arg Pro Leu Gln Gln Pro385 390 395 400gca cca gct cct cct ttt aat tta cgt agc tta gag gga gta gaa ttc 1248Ala Pro Ala Pro Pro Phe Asn Leu Arg Ser Leu Glu Gly Val Glu Phe405 410 415cac act cct aca ggt agt ttt atg tat cgt gaa aga gga tcg gta gat 1296His Thr Pro Thr Gly Ser Phe Met Tyr Arg Glu Arg Gly Ser Val Asp420 425 430tct ttt aat gag ttg ccg cct ttt aat cca gtt ggg tta cct cat aag 1344Ser Phe Asn Glu Leu Pro Pro Phe Asn Pro Val Gly Leu Pro His Lys435 440 445gta tac agt cac cgt tta tgt cat gca acg ttt gtt cgt aaa tct ggg 1392
Val Tyr Ser His Arg Leu Cys His Ala Thr Phe Val Arg Lys Ser Gly450 455 460acc cct tat tta aca aca ggt gcc atc ttt tct tgg aca cat cgt agt 1440Thr Pro Tyr Leu Thr Thr Gly Ala Ile Phe Ser Trp Thr His Arg Ser465 470 475 480gct gaa gaa acc aat aca att gaa tca aat att att acg caa atc ccg 1488Ala Glu Glu Thr Asn Thr Ile Glu Ser Asn Ile Ile Thr Gln Ile Pro485 490 495tta gta aaa gca tat caa att ggg tca ggc act act gta agg aaa gga 1536Leu Val Lys Ala Tyr Gln Ile Gly Ser Gly Thr Thr Val Arg Lys Gly500 505 510cca gga ttc aca gga ggg gat ata ctt cga aga aca ggt cct gga aca 1584Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Gly Pro Gly Thr515 520 525ttt gga gat atg aga ata aat att aat gca cca tta tct caa aga tat 1632Phe Gly Asp Met Arg Ile Asn Ile Asn Ala Pro Leu Ser Gln Arg Tyr530 535 540cgt gta agg att cgt tat gct tct acg aca gat tta caa ttt gtc acg 1680Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asp Leu Gln Phe Val Thr545 550 555 560agt att aat ggg acc acc att aat att ggt aac ttc ccg aaa act att 1728Ser Ile Asn Gly Thr Thr Ile Asn Ile Gly Asn Phe Pro Lys Thr Ile565 570 575aat aat cta aat act tta ggt tct gag ggc tat aga aca gta tcg ttt 1776Asn Asn Leu Asn Thr Leu Gly Ser Glu Gly Tyr Arg Thr Val Ser Phe580 585 590agt act cca ttt agt ttc tca aat gca caa agc ata ttt aga tta ggt 1824Ser Thr Pro Phe Ser Phe Ser Asn Ala Gln Ser Ile Phe Arg Leu Gly595 600 605ata caa gca ttt tct gga gtt caa gaa gtt tat gtg gat aaa att gaa 1872Ile Gln Ala Phe Ser Gly Val Gln Glu Val Tyr Val Asp Lys Ile Glu610 615 620ttt att cct gtt gaa tag 1890Phe Ile Pro Val Glu *625<210>3<211>629<212>PRT<213>Bacillus thuringiensis<400>3Met Ser Glu Leu Lys Gly Lys Phe Lys Lys Ser Thr Asn Arg Thr Cys1 5 10 15Cys Leu Leu Lys Ile Ile Asn Ile Gly Gly Arg Gly Met Asn Ser Lys20 25 30Glu His Asp Tyr Leu Lys Val Cys Asn Asp Leu Ser Asp Ala Asn Ile35 40 45Asn Met Glu Arg Phe Asp Lys Asn Asp Ala Leu Glu Ile Gly Met Ser50 55 60Ile Val Ser Glu Leu Ile Gly Met Ile Pro Gly Gly Thr Ala Leu Gln65 70 75 80Phe Val Phe Asn Gln Leu Trp Ser Arg Leu Gly Asp Ser Gly Trp Asn85 90 95Ala Phe Met Glu His Val Glu Glu Leu Ile Asp Thr Lys Ile Glu Gly100 105 110
Tyr Ala Lys Asn Lys Ala Leu Ser Glu Leu Ala Gly Ile Gln Arg Asn115 120 125Leu Glu Thr Tyr Ile Gln Leu Arg Asn Glu Trp Glu Asn Asp Ile Glu130 135 140Asn Ser Lys Ala Gln Gly Lys Val Ala Asn Tyr Tyr Glu Ser Leu Glu145 150 155 160Gln Ala Val Glu Arg Ser Met Pro Gln Phe Ala Val Glu Asn Phe Glu165 170 175Val Pro Leu Leu Thr Val Tyr Val Gln Ala Ala Asn Leu His Leu Leu180 185 190Leu Leu Arg Asp Val Ser Val Tyr Gly Lys Cys Trp Gly Trp Ser Glu195 200 205Gln Lys Ile Lys Ile Tyr Tyr Asp Lys Gln Ile Lys Tyr Thr His Glu210 215 220Tyr Thr Asn His Cys Val Asn Trp Tyr Asn Lys Gly Leu Glu Arg Leu225 230 235 240Lys Asn Lys Gly Ser Ser Tyr Gln Asp Trp Tyr Asn Tyr Asn Arg Phe245 250 255Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Val Ala Leu Phe Pro260 265 270His Tyr Asp Val Gln Thr Tyr Pro Ile Thr Thr Val Ala Gln Leu Thr275 280 285Arg Glu Val Tyr Thr Asp Pro Leu Leu Asn Phe Asn Pro Lys Leu His290 295 300Ser Val Ser Gln Leu Pro Ser Phe Ser Asp Met Glu Asn Ala Thr Ile305 310 315 320Arg Thr Pro His Leu Met Glu Phe Leu Arg Met Leu Thr Ile Tyr Thr325 330 335Asp Trp Tyr Ser Val Gly Arg Asn Tyr Tyr Trp Gly Gly His Arg Val340 345 350Thr Ser Tyr His Val Gly Gly Glu Asn Ile Arg Ser Pro Leu Tyr Gly355 360 365Arg Glu Ala Asn Gln Glu Val Pro Arg Asp Phe Tyr Phe Tyr Gly Pro370 375 380Val Phe Lys Thr Leu Ser Lys Pro Thr Leu Arg Pro Leu Gln Gln Pro385 390 395 400Ala Pro Ala Pro Pro Phe Asn Leu Arg Ser Leu Glu Gly Val Glu Phe405 410 415His Thr Pro Thr Gly Ser Phe Met Tyr Arg Glu Arg Gly Ser Val Asp420 425 430Ser Phe Asn Glu Leu Pro Pro Phe Asn Pro Val Gly Leu Pro His Lys435 440 445Val Tyr Ser His Arg Leu Cys His Ala Thr Phe Val Arg Lys Ser Gly450 455 460Thr Pro Tyr Leu Thr Thr Gly Ala Ile Phe Ser Trp Thr His Arg Ser465 470 475 480Ala Glu Glu Thr Asn Thr Ile Glu Ser Asn Ile Ile Thr Gln Ile Pro485 490 495Leu Val Lys Ala Tyr Gln Ile Gly Ser Gly Thr Thr Val Arg Lys Gly500 505 510Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Gly Pro Gly Thr515 520 525Phe Gly Asp Met Arg Ile Asn Ile Asn Ala Pro Leu Ser Gln Arg Tyr530 535 540Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asp Leu Gln Phe Val Thr545 550 555 560Ser Ile Asn Gly Thr Thr Ile Asn Ile Gly Asn Phe Pro Lys Thr Ile565 570 575Asn Asn Leu Asn Thr Leu Gly Ser Glu Gly Tyr Arg Thr Val Ser Phe580 585 590Ser Thr Pro Phe Ser Phe Ser Asn Ala Gln Ser Ile Phe Arg Leu Gly595 600 605Ile Gln Ala Phe Ser Gly Val Gln Glu Val Tyr Val Asp Lys Ile Glu610 615 620Phe Ile Pro Val Glu625
<210>4<211>1806<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(1806)<400>4atg aat tca aag gaa cat gat tat cta aaa gtt tgt aat gat tta agt 48Met Asn Ser Lys Glu His Asp Tyr Leu Lys Val Cys Asn Asp Leu Ser1 5 10 15gac gcc aat att aat atg gaa cgg ttt gat aag aat gat gca ctg gaa 96Asp Ala Asn Ile Asn Met Glu Arg Phe Asp Lys Asn Asp Ala Leu Glu20 25 30att ggt atg tcc att gta tct gaa ctt att ggt atg att cca ggc gga 144Ile Gly Met Ser Ile Val Ser Glu Leu Ile Gly Met Ile Pro Gly Gly35 40 45aca gct ttg caa ttt gtg ttt aat caa ttg tgg tct cgt tta ggt gat 192Thr Ala Leu Gln Phe Val Phe Asn Gln Leu Trp Ser Arg Leu Gly Asp50 55 60tct gga tgg aat gcg ttc atg gaa cat gtg gag gaa tta att gat act 240Ser Gly Trp Asn Ala Phe Met Glu His Val Glu Glu Leu Ile Asp Thr65 70 75 80aaa ata gaa ggg tat gca aaa aat aaa gcc tta tct gaa tta gca ggt 288Lys Ile Glu Gly Tyr Ala Lys Asn Lys Ala Leu Ser Glu Leu Ala Gly85 90 95ata caa aga aac ctt gaa aca tat ata caa tta cgt aat gaa tgg gaa 336Ile Gln Arg Asn Leu Glu Thr Tyr Ile Gln Leu Arg Asn Glu Trp Glu100 105 110aat gat att gaa aac tca aag gct caa ggt aag gta gct aat tac tat 384Asn Asp Ile Glu Asn Ser Lys Ala Gln Gly Lys Val Ala Asn Tyr Tyr115 120 125gaa agt ctt gag cag gcg gtt gaa agg agt atg cct caa ttt gca gtg 432Glu Ser Leu Glu Gln Ala Val Glu Arg Ser Met Pro Gln Phe Ala Val130 135 140gag aat ttt gaa gta cca ctt tta act gtc tat gtg caa gct gct aat 480Glu Asn Phe Glu Val Pro Leu Leu Thr Val Tyr Val Gln Ala Ala Asn145 150 155 160ctt cat tta tta tta tta aga gat gtt tca gtt tat gga aag tgt tgg 528Leu His Leu Leu Leu Leu Arg Asp Val Ser Val Tyr Gly Lys Cys Trp165 170 175gga tgg tcg gag cag aaa att aaa att tat tat gat aaa cag att aag 576Gly Trp Ser Glu Gln Lys Ile Lys Ile Tyr Tyr Asp Lys Gln Ile Lys180 185 190tat acc cat gaa tac aca aat cat tgt gta aat tgg tat aat aaa gga 624Tyr Thr His Glu Tyr Thr Asn His Cys Val Asn Trp Tyr Asn Lys Gly195 200 205ctt gag aga tta aaa aat aaa ggt tct tct tat caa gat tgg tac aat 672Leu Glu Arg Leu Lys Asn Lys Gly Ser Ser Tyr Gln Asp Trp Tyr Asn210 215 220tat aat cgt ttc cgt aga gaa atg act ctt act gtt tta gat atc gtt 720Tyr Asn Arg Phe Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Val
225 230 235 240gct tta ttc ccg cac tat gat gta caa act tat cca ata aca acc gtt 768Ala Leu Phe Pro His Tyr Asp Val Gln Thr Tyr Pro Ile Thr Thr Val245 250 255gct cag cta aca agg gaa gtt tat acg gat cct tta ctt aat ttt aat 816Ala Gln Leu Thr Arg Glu Val Tyr Thr Asp Pro Leu Leu Asn Phe Asn260 265 270cct aaa tta cat tct gtg tct caa tta cct agt ttt agt gac atg gaa 864Pro Lys Leu His Ser Val Ser Gln Leu Pro Ser Phe Ser Asp Met Glu275 280 285aat gca aca att aga act cca cat ctg atg gaa ttt tta aga atg cta 912Asn Ala Thr Ile Arg Thr Pro His Leu Met Glu Phe Leu Arg Met Leu290 295 300aca att tat aca gat tgg tat agt gtg gga aga aac tat tat tgg gga 960Thr Ile Tyr Thr Asp Trp Tyr Ser Val Gly Arg Asn Tyr Tyr Trp Gly305 310 315 320gga cat cgc gtg acg tct tac cat gta gga gga gag aat ata aga tca 1008Gly His Arg Val Thr Ser Tyr His Val Gly Gly Glu Asn Ile Arg Ser325 330 335cct cta tat ggt aga gag gca aat caa gag gtt cct aga gat ttt tat 1056Pro Leu Tyr Gly Arg Glu Ala Asn Gln Glu Val Pro Arg Asp Phe Tyr340 345 350ttt tat gga ccc gtt ttt aag acg tta tca aag ccg act cta aga cca 1104Phe Tyr Gly Pro Val Phe Lys Thr Leu Ser Lys Pro Thr Leu Arg Pro355 360 365tta cag cag cct gca cca gct cct cct ttt aat tta cgt agc tta gag 1152Leu Gln Gln Pro Ala Pro Ala Pro Pro Phe Asn Leu Arg Ser Leu Glu370 375 380gga gta gaa ttc cac act cct aca ggt agt ttt atg tat cgt gaa aga 1200Gly Val Glu Phe His Thr Pro Thr Gly Ser Phe Met Tyr Arg Glu Arg385 390 395 400gga tcg gta gat tct ttt aat gag ttg ccg cct ttt aat cca gtt ggg 1248Gly Ser Val Asp Ser Phe Asn Glu Leu Pro Pro Phe Asn Pro Val Gly405 410 415tta cct cat aag gta tac agt cac cgt tta tgt cat gca acg ttt gtt 1296Leu Pro His Lys Val Tyr Ser His Arg Leu Cys His Ala Thr Phe Val420 425 430cgt aaa tct ggg acc cct tat tta aca aca ggt gcc atc ttt tct tgg 1344Arg Lys Ser Gly Thr Pro Tyr Leu Thr Thr Gly Ala Ile Phe Ser Trp435 440 445aca cat cgt agt gct gaa gaa acc aat aca att gaa tca aat att att 1392Thr His Arg Ser Ala Glu Glu Thr Asn Thr Ile Glu Ser Asn Ile Ile450 455 460acg caa atc ccg tta gta aaa gca tat caa att ggg tca ggc act act 1440Thr Gln Ile Pro Leu Val Lys Ala Tyr Gln Ile Gly Ser Gly Thr Thr465 470 475 480gta agg aaa gga cca gga ttc aca gga ggg gat ata ctt cga aga aca 1488Val Arg Lys Gly Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr485 490 495ggt cct gga aca ttt gga gat atg aga ata aat att aat gca cca tta 1536Gly Pro Gly Thr Phe Gly Asp Met Arg Ile Asn Ile Asn Ala Pro Leu
500 505 510tct caa aga tat cgt gta agg att cgt tat gct tct acg aca gat tta 1584Ser Gln Arg Tyr Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asp Leu515 520 525caa ttt gtc acg agt att aat ggg acc acc att aat att ggt aac ttc 1632Gln Phe Val Thr Ser Ile Asn Gly Thr Thr Ile Asn Ile Gly Asn Phe530 535 540ccg aaa act att aat aat cta aat act tta ggt tct gag ggc tat aga 1680Pro Lys Thr Ile Asn Asn Leu Asn Thr Leu Gly Ser Glu Gly Tyr Arg545 550 555 560aca gta tcg ttt agt act cca ttt agt ttc tca aat gca caa agc ata 1728Thr Val Ser Phe Ser Thr Pro Phe Ser Phe Ser Asn Ala Gln Ser Ile565 570 575ttt aga tta ggt ata caa gca ttt tct gga gtt caa gaa gtt tat gtg 1776Phe Arg Leu Gly Ile Gln Ala Phe Ser Gly Val Gln Glu Val Tyr Val580 585 590gat aaa att gaa ttt att cct gtt gaa tag 1806Asp Lys Ile Glu Phe Ile Pro Val Glu *595 600<210>5<211>601<212>PRT<213>Bacillus thuringiensis<400>5Met Asn Ser Lys Glu His Asp Tyr Leu Lys Val Cys Asn Asp Leu Ser1 5 10 15Asp Ala Asn Ile Asn Met Glu Arg Phe Asp Lys Asn Asp Ala Leu Glu20 25 30Ile Gly Met Ser Ile Val Ser Glu Leu Ile Gly Met Ile Pro Gly Gly35 40 45Thr Ala Leu Gln Phe Val Phe Asn Gln Leu Trp Ser Arg Leu Gly Asp50 55 60Ser Gly Trp Asn Ala Phe Met Glu His Val Glu Glu Leu Ile Asp Thr65 70 75 80Lys Ile Glu Gly Tyr Ala Lys Asn Lys Ala Leu Ser Glu Leu Ala Gly85 90 95Ile Gln Arg Asn Leu Glu Thr Tyr Ile Gln Leu Arg Asn Glu Trp Glu100 105 110Asn Asp Ile Glu Asn Ser Lys Ala Gln Gly Lys Val Ala Asn Tyr Tyr115 120 125Glu Ser Leu Glu Gln Ala Val Glu Arg Ser Met Pro Gln Phe Ala Val130 135 140Glu Asn Phe Glu Val Pro Leu Leu Thr Val Tyr Val Gln Ala Ala Asn145 150 155 160Leu His Leu Leu Leu Leu Arg Asp Val Ser Val Tyr Gly Lys Cys Trp165 170 175Gly Trp Ser Glu Gln Lys Ile Lys Ile Tyr Tyr Asp Lys Gln Ile Lys180 185 190Tyr Thr His Glu Tyr Thr Asn His Cys Val Asn Trp Tyr Asn Lys Gly195 200 205Leu Glu Arg Leu Lys Asn Lys Gly Ser Ser Tyr Gln Asp Trp Tyr Asn210 215 220Tyr Asn Arg Phe Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Val225 230 235 240Ala Leu Phe Pro His Tyr Asp Val Gln Thr Tyr Pro Ile Thr Thr Val245 250 255Ala Gln Leu Thr Arg Glu Val Tyr Thr Asp Pro Leu Leu Asn Phe Asn260 265 270Pro Lys Leu His Ser Val Ser Gln Leu Pro Ser Phe Ser Asp Met Glu
275 280 285Asn Ala Thr Ile Arg Thr Pro His Leu Met Glu Phe Leu Arg Met Leu290 295 300Thr Ile Tyr Thr Asp Trp Tyr Ser Val Gly Arg Asn Tyr Tyr Trp Gly305 310 315 320Gly His Arg Val Thr Ser Tyr His Val Gly Gly Glu Asn Ile Arg Ser325 330 335Pro Leu Tyr Gly Arg Glu Ala Asn Gln Glu Val Pro Arg Asp Phe Tyr340 345 350Phe Tyr Gly Pro Val Phe Lys Thr Leu Ser Lys Pro Thr Leu Arg Pro355 360 365Leu Gln Gln Pro Ala Pro Ala Pro Pro Phe Asn Leu Arg Ser Leu Glu370 375 380Gly Val Glu Phe His Thr Pro Thr Gly Ser Phe Met Tyr Arg Glu Arg385 390 395 400Gly Ser Val Asp Ser Phe Asn Glu Leu Pro Pro Phe Asn Pro Val Gly405 410 415Leu Pro His Lys Val Tyr Ser His Arg Leu Cys His Ala Thr Phe Val420 425 430Arg Lys Ser Gly Thr Pro Tyr Leu Thr Thr Gly Ala Ile Phe Ser Trp435 440 445Thr His Arg Ser Ala Glu Glu Thr Asn Thr Ile Glu Ser Asn Ile Ile450 455 460Thr Gln Ile Pro Leu Val Lys Ala Tyr Gln Ile Gly Ser Gly Thr Thr465 470 475 480Val Arg Lys Gly Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr485 490 495Gly Pro Gly Thr Phe Gly Asp Met Arg Ile Asn Ile Asn Ala Pro Leu500 505 510Ser Gln Arg Tyr Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asp Leu515 520 525Gln Phe Val Thr Ser Ile Asn Gly Thr Thr Ile Asn Ile Gly Asn Phe530 535 540Pro Lys Thr Ile Asn Asn Leu Asn Thr Leu Gly Ser Glu Gly Tyr Arg545 550 555 560Thr Val Ser Phe Ser Thr Pro Phe Ser Phe Ser Asn Ala Gln Ser Ile565 570 575Phe Arg Leu Gly Ile Gln Ala Phe Ser Gly Val Gln Glu Val Tyr Val580 585 590Asp Lys Ile Glu Phe Ile Pro Val Glu595 600<210>6<211>2208<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2208)<400>6atg aat caa aat aac gat aat aac gaa tat gaa att att gat tcg cat 48Met Asn Gln Asn Asn Asp Asn Asn Glu Tyr Glu Ile Ile Asp Ser His1 5 10 15acc tca cct tat ttt ccg aac aga aac agt aat gat tct aga tac cct 96Thr Ser Pro Tyr Phe Pro Asn Arg Asn Ser Asn Asp Ser Arg Tyr Pro20 25 30tac aca aat aat cca aat caa cca tta caa aac aca aat tac aaa gag 144Tyr Thr Asn Asn Pro Asn Gln Pro Leu Gln Asn Thr Asn Tyr Lys Glu35 40 45tgg ctc aat atg tgt caa ggg aat aca caa tat ggt gat aat ttc gag 192Trp Leu Asn Met Cys Gln Gly Asn Thr Gln Tyr Gly Asp Asn Phe Glu50 55 60
aca ttt gct agt gct gat aca att gct gca gtt agt gca ggt act att 240Thr Phe Ala Ser Ala Asp Thr Ile Ala Ala Val Ser Ala Gly Thr Ile65 70 75 80gta tcc ggt act ctg tta gcc ggt ata ggt ggg ctc act tct ata tcc 288Val Ser Gly Thr Leu Leu Ala Gly Ile Gly Gly Leu Thr Ser Ile Ser85 90 95gga ccg ata gga ata ata ggt gct ata ata ata tct ttt ggt acc cta 336Gly Pro Ile Gly Ile Ile Gly Ala Ile Ile Ile Ser Phe Gly Thr Leu100 105 110atc act gtc ttt tgg ccc gcg gga gaa caa gac aaa aca gta tgg aca 384Ile Thr Val Phe Trp Pro Ala Gly Glu Gln Asp Lys Thr Val Trp Thr115 120 125caa ttt att aaa atg gga gaa att ttt gtt gat aca ccg tta aca gaa 432Gln Phe Ile Lys Met Gly Glu Ile Phe Val Asp Thr Pro Leu Thr Glu130 135 140agc ata aaa cag cta aag tta caa act tta gaa gga ttt aga caa ata 480Ser Ile Lys Gln Leu Lys Leu Gln Thr Leu Glu Gly Phe Arg Gln Ile145 150 155 160tta caa agc tat aat aca gca tta gat gat tgg aga aaa tta aaa aga 528Leu Gln Ser Tyr Asn Thr Ala Leu Asp Asp Trp Arg Lys Leu Lys Arg165 170 175cta caa gct cct gga tta cca cca tca tca gca tta caa caa gct gcc 576Leu Gln Ala Pro Gly Leu Pro Pro Ser Ser Ala Leu Gln Gln Ala Ala180 185 190ttg act ctt aaa ata cga ttt gag aat gtt cac aat gat ttt att cga 624Leu Thr Leu Lys Ile Arg Phe Glu Asn Val His Asn Asp Phe Ile Arg195 200 205gaa ata cct ggt ttc caa ctt gaa act tat aaa acg cta tta cta cct 672Glu Ile Pro Gly Phe Gln Leu Glu Thr Tyr Lys Thr Leu Leu Leu Pro210 215 220att tat gcg caa gct gct aat ttt cat tta aat tta tta caa caa ggt 720Ile Tyr Ala Gln Ala Ala Asn Phe His Leu Asn Leu Leu Gln Gln Gly225 230 235 240gct gaa ttg gct gat gaa tgg aat gca gat ata cat cct tca caa att 768Ala Glu Leu Ala Asp Glu Trp Asn Ala Asp Ile His Pro Ser Gln Ile245 250 255gaa cct aat gct gga aca tca gat gac tat tat aaa ctt tta aaa gaa 816Glu Pro Asn Ala Gly Thr Ser Asp Asp Tyr Tyr Lys Leu Leu Lys Glu260 265 270aat ata cct aaa tat agt aac tat tgt gca aat acc tat aga aca gga 864Asn Ile Pro Lys Tyr Ser Asn Tyr Cys Ala Asn Thr Tyr Arg Thr Gly275 280 285cta aaa aat ctt aga gac gaa cca aat atg aaa tgg agt ata ttt aat 912Leu Lys Asn Leu Arg Asp Glu Pro Asn Met Lys Trp Ser Ile Phe Asn290 295 300gac tat cga aga tat atg acc att act gta tta gat acc atc tct caa 960Asp Tyr Arg Arg Tyr Met Thr Ile Thr Val Leu Asp Thr Ile Ser Gln305 310 315 320ttt tct tta tat gat ata aaa aga tat aga gat tca ata gga gga ata 1008Phe Ser Leu Tyr Asp Ile Lys Arg Tyr Arg Asp Ser Ile Gly Gly Ile325 330 335
gaa gta aaa ggc att aag aat gaa ctc aca aga gaa att tat aca act 1056Glu Val Lys Gly Ile Lys Asn Glu Leu Thr Arg Glu Ile Tyr Thr Thr340 345 350gaa ata aat ttt gat cgt ctt cct caa ctt aga gtt caa ccc aat cta 1104Glu Ile Asn Phe Asp Arg Leu Pro Gln Leu Arg Val Gln Pro Asn Leu355 360 365gct acg atg gaa tat aat tta aca cgt gca agt ttt aaa tta ttt tca 1152Ala Thr Met Glu Tyr Asn Leu Thr Arg Ala Ser Phe Lys Leu Phe Ser370 375 380ttt tta gaa caa ttt att ttt tat aca gaa aat aca aat ttc ggg aat 1200Phe Leu Glu Gln Phe Ile Phe Tyr Thr Glu Asn Thr Asn Phe Gly Asn385 390 395 400cgt tta gtt ggt att tct aat cgt gat gca cct act tat agc aat act 1248Arg Leu Val Gly Ile Ser Asn Arg Asp Ala Pro Thr Tyr Ser Asn Thr405 410 415ata act gaa act tta tat gga gaa aga aca ggt tca ccc aca aca aaa 1296Ile Thr Glu Thr Leu Tyr Gly Glu Arg Thr Gly Ser Pro Thr Thr Lys420 425 430aca ata aga cca ttt gaa tct tat aaa gtt tca att gta act gat aga 1344Thr Ile Arg Pro Phe Glu Ser Tyr Lys Val Ser Ile Val Thr Asp Arg435 440 445caa tca cct cct gtt tcc cct att caa cca cac ttt ata att aat caa 1392Gln Ser Pro Pro Val Ser Pro Ile Gln Pro His Phe Ile Ile Asn Gln450 455 460att gaa ctt tat tta aat ggc tca tct aac aac aca ctc aaa tat tca 1440Ile Glu Leu Tyr Leu Asn Gly Ser Ser Asn Asn Thr Leu Lys Tyr Ser465 470 475 480gca gga ggg tct tta tct aat tat caa aac aca act ttt ttt caa ttt 1488Ala Gly Gly Ser Leu Ser Asn Tyr Gln Asn Thr Thr Phe Phe Gln Phe485 490 495cct aga aaa aaa gac tgc aat cta gtt att gat cca ggt tgt tca cca 1536Pro Arg Lys Lys Asp Cys Asn Leu Val Ile Asp Pro Gly Cys Ser Pro500 505 510aac ttt aat aac tat agt cat att tta tcc cat ttt tca tta ttt act 1584Asn Phe Asn Asn Tyr Ser His Ile Leu Ser His Phe Ser Leu Phe Thr515 520 525tat tcc tat gtg att gga tta cag cta caa ata tta gat aca ggt gta 1632Tyr Ser Tyr Val Ile Gly Leu Gln Leu Gln Ile Leu Asp Thr Gly Val530 535 540tta gga tgg aca cac agt agt gtt gat aga tat aat gca ata tca gat 1680Leu Gly Trp Thr His Ser Ser Val Asp Arg Tyr Asn Ala Ile Ser Asp545 550 555 560aaa ata att aca atg atc cca gca atc aaa ggt aac aat ctt gat aca 1728Lys Ile Ile Thr Met Ile Pro Ala Ile Lys Gly Asn Asn Leu Asp Thr565 570 575aac tct aag gta att gaa gga cct ggt cat aca gga gga aac ttg gtt 1776Asn Ser Lys Val Ile Glu Gly Pro Gly His Thr Gly Gly Asn Leu Val580 585 590tat tta caa agt caa ggg cgt tta gaa att aca tgt gaa act cct aat 1824Tyr Leu Gln Ser Gln Gly Arg Leu Glu Ile Thr Cys Glu Thr Pro Asn595 600 605
tct aca caa tct tat ttc att aga ctt cga tat gct aca aat ggt gct 1872Ser Thr Gln Ser Tyr Phe Ile Arg Leu Arg Tyr Ala Thr Asn Gly Ala610 615 620gga aat act ctt cct aat ata tct ctt aca ata cca gga gta ata gga 1920Gly Asn Thr Leu Pro Asn Ile Ser Leu Thr Ile Pro Gly Val Ile Gly625 630 635 640ata cca cct caa cga ctc aac aac act ttt tct ggt aca aat tat aat 1968Ile Pro Pro Gln Arg Leu Asn Asn Thr Phe Ser Gly Thr Asn Tyr Asn645 650 655aat tta caa tac gga gat ttt ggg tat ttc caa ttt cca agt aca gta 2016Asn Leu Gln Tyr Gly Asp Phe Gly Tyr Phe Gln Phe Pro Ser Thr Val660 665 670aca tta cct tta aat cga aac ata cca ttt ata ttt aat cgt gca gat 2064Thr Leu Pro Leu Asn Arg Asn Ile Pro Phe Ile Phe Asn Arg Ala Asp675 680 685gta tca aat tca att tta atc att gat aaa att gaa ttt ata cca att 2112Val Ser Asn Ser Ile Leu Ile Ile Asp Lys Ile Glu Phe Ile Pro Ile690 695 700act tcc tct atg cac caa aat aga gaa aaa caa aaa tta gaa act atc 2160Thr Ser Ser Met His Gln Asn Arg Glu Lys Gln Lys Leu Glu Thr Ile705 710 715 720caa aca aaa ata aat aca ttt ttc aca aat cat aca aaa aca ctt tga 2208Gln Thr Lys Ile Asn Thr Phe Phe Thr Asn His Thr Lys Thr Leu *725 730 735<210>7<211>735<212>PRT<213>Bacillus thuringiensis<400>7Met Asn Gln Asn Asn Asp Asn Asn Glu Tyr Glu Ile Ile Asp Ser His1 5 10 15Thr Ser Pro Tyr Phe Pro Asn Arg Asn Ser Asn Asp Ser Arg Tyr Pro20 25 30Tyr Thr Asn Asn Pro Asn Gln Pro Leu Gln Asn Thr Asn Tyr Lys Glu35 40 45Trp Leu Asn Met Cys Gln Gly Asn Thr Gln Tyr Gly Asp Asn Phe Glu50 55 60Thr Phe Ala Ser Ala Asp Thr Ile Ala Ala Val Ser Ala Gly Thr Ile65 70 75 80Val Ser Gly Thr Leu Leu Ala Gly Ile Gly Gly Leu Thr Ser Ile Ser85 90 95Gly Pro Ile Gly Ile Ile Gly Ala Ile Ile Ile Ser Phe Gly Thr Leu100 105 110Ile Thr Val Phe Trp Pro Ala Gly Glu Gln Asp Lys Thr Val Trp Thr115 120 125Gln Phe Ile Lys Met Gly Glu Ile Phe Val Asp Thr Pro Leu Thr Glu130 135 140Ser Ile Lys Gln Leu Lys Leu Gln Thr Leu Glu Gly Phe Arg Gln Ile145 150 155 160Leu Gln Ser Tyr Asn Thr Ala Leu Asp Asp Trp Arg Lys Leu Lys Arg165 170 175Leu Gln Ala Pro Gly Leu Pro Pro Ser Ser Ala Leu Gln Gln Ala Ala180 185 190Leu Thr Leu Lys Ile Arg Phe Glu Asn Val His Asn Asp Phe Ile Arg195 200 205Glu Ile Pro Gly Phe Gln Leu Glu Thr Tyr Lys Thr Leu Leu Leu Pro
210 215 220Ile Tyr Ala Gln Ala Ala Asn Phe His Leu Asn Leu Leu Gln Gln Gly225 230 235 240Ala Glu Leu Ala Asp Glu Trp Asn Ala Asp Ile His Pro Ser Gln Ile245 250 255Glu Pro Asn Ala Gly Thr Ser Asp Asp Tyr Tyr Lys Leu Leu Lys Glu260 265 270Asn Ile Pro Lys Tyr Ser Asn Tyr Cys Ala Asn Thr Tyr Arg Thr Gly275 280 285Leu Lys Asn Leu Arg Asp Glu Pro Asn Met Lys Trp Ser Ile Phe Asn290 295 300Asp Tyr Arg Arg Tyr Met Thr Ile Thr Val Leu Asp Thr Ile Ser Gln305 310 315 320Phe Ser Leu Tyr Asp Ile Lys Arg Tyr Arg Asp Ser Ile Gly Gly Ile325 330 335Glu Val Lys Gly Ile Lys Asn Glu Leu Thr Arg Glu Ile Tyr Thr Thr340 345 350Glu Ile Asn Phe Asp Arg Leu Pro Gln Leu Arg Val Gln Pro Asn Leu355 360 365Ala Thr Met Glu Tyr Asn Leu Thr Arg Ala Ser Phe Lys Leu Phe Ser370 375 380Phe Leu Glu Gln Phe Ile Phe Tyr Thr Glu Asn Thr Asn Phe Gly Asn385 390 395 400Arg Leu Val Gly Ile Ser Asn Arg Asp Ala Pro Thr Tyr Ser Asn Thr405 410 415Ile Thr Glu Thr Leu Tyr Gly Glu Arg Thr Gly Ser Pro Thr Thr Lys420 425 430Thr Ile Arg Pro Phe Glu Ser Tyr Lys Val Ser Ile Val Thr Asp Arg435 440 445Gln Ser Pro Pro Val Ser Pro Ile Gln Pro His Phe Ile Ile Asn Gln450 455 460Ile Glu Leu Tyr Leu Asn Gly Ser Ser Asn Asn Thr Leu Lys Tyr Ser465 470 475 480Ala Gly Gly Ser Leu Ser Asn Tyr Gln Asn Thr Thr Phe Phe Gln Phe485 490 495Pro Arg Lys Lys Asp Cys Asn Leu Val Ile Asp Pro Gly Cys Ser Pro500 505 510Asn Phe Asn Asn Tyr Ser His Ile Leu Ser His Phe Ser Leu Phe Thr515 520 525Tyr Ser Tyr Val Ile Gly Leu Gln Leu Gln Ile Leu Asp Thr Gly Val530 535 540Leu Gly Trp Thr His Ser Ser Val Asp Arg Tyr Asn Ala Ile Ser Asp545 550 555 560Lys Ile Ile Thr Met Ile Pro Ala Ile Lys Gly Asn Asn Leu Asp Thr565 570 575Asn Ser Lys Val Ile Glu Gly Pro Gly His Thr Gly Gly Asn Leu Val580 585 590Tyr Leu Gln Ser Gln Gly Arg Leu Glu Ile Thr Cys Glu Thr Pro Asn595 600 605Ser Thr Gln Ser Tyr Phe Ile Arg Leu Arg Tyr Ala Thr Asn Gly Ala610 615 620Gly Asn Thr Leu Pro Asn Ile Ser Leu Thr Ile Pro Gly Val Ile Gly625 630 635 640Ile Pro Pro Gln Arg Leu Asn Asn Thr Phe Ser Gly Thr Asn Tyr Asn645 650 655Asn Leu Gln Tyr Gly Asp Phe Gly Tyr Phe Gln Phe Pro Ser Thr Val660 665 670Thr Leu Pro Leu Asn Arg Asn Ile Pro Phe Ile Phe Asn Arg Ala Asp675 680 685Val Ser Asn Ser Ile Leu Ile Ile Asp Lys Ile Glu Phe Ile Pro Ile690 695 700Thr Ser Ser Met His Gln Asn Arg Glu Lys Gln Lys Leu Glu Thr Ile705 710 715 720Gln Thr Lys Ile Asn Thr Phe Phe Thr Asn His Thr Lys Thr Leu725 730 735<210>8
<211>2235<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2235)<400>8gtg aat caa aat aat aat aat gaa tat gag att atc gat tca aag aat 48Met Asn Gln Asn Asn Asn Asn Glu Tyr Glu Ile Ile Asp Ser Lys Asn1 5 10 15tta tct tat cct tct aac aga aat att gat cat tct aga tac cct tac 96Leu Ser Tyr Pro Ser Asn Arg Asn Ile Asp His Ser Arg Tyr Pro Tyr20 25 30aca aat aat cca aat caa cca tta caa aac aca aat tac aaa gag tgg 144Thr Asn Asn Pro Asn Gln Pro Leu Gln Asn Thr Asn Tyr Lys Glu Trp35 40 45ctc aat atg tgt caa ggg aat aca caa tat ggt gat aat ttc gag aca 192Leu Asn Met Cys Gln Gly Asn Thr Gln Tyr Gly Asp Asn Phe Glu Thr50 55 60ttt gct agt gct gat aca att gct gca gtt agt gca ggt act att gta 240Phe Ala Ser Ala Asp Thr Ile Ala Ala Val Ser Ala Gly Thr Ile Val65 70 75 80tcc ggt act ctg tta gcc ggt ata ggt ggg ctc act tct ata tcc gga 288Ser Gly Thr Leu Leu Ala Gly Ile Gly Gly Leu Thr Ser Ile Ser Gly85 90 95ccg ata gga ata ata ggt gct ata ata ata tct ttt ggt acc cta atc 336Pro Ile Gly Ile Ile Gly Ala Ile Ile Ile Ser Phe Gly Thr Leu Ile100 105 110act gtc ttt tgg ccc gcg gga gaa caa gac aaa aca gta tgg aca caa 384Thr Val Phe Trp Pro Ala Gly Glu Gln Asp Lys Thr Val Trp Thr Gln115 120 125ttt att aaa atg gga gaa att ttt gtt gat aca ccg tta aca gaa agc 432Phe Ile Lys Met Gly Glu Ile Phe Val Asp Thr Pro Leu Thr Glu Ser130 135 140ata aaa cag cta aag tta caa act tta gaa gga ttt aga caa ata tta 480Ile Lys Gln Leu Lys Leu Gln Thr Leu Glu Gly Phe Arg Gln Ile Leu145 150 155 160caa agc tat aat aca gca tta gat gat tgg aga aaa tta aaa aga cta 528Gln Ser Tyr Asn Thr Ala Leu Asp Asp Trp Arg Lys Leu Lys Arg Leu165 170 175caa gct cct gga tta cca cca tca tca gca tta caa caa gct gcc ttg 576Gln Ala Pro Gly Leu Pro Pro Ser Ser Ala Leu Gln Gln Ala Ala Leu180 185 190act ctt aaa ata cga ttt gag aat gtt cac aat gat ttt att cga gaa 624Thr Leu Lys Ile Arg Phe Glu Asn Val His Asn Asp Phe Ile Arg Glu195 200 205ata cct ggt ttc caa ctt gaa act tat aaa acg cta tta cta cct att 672Ile Pro Gly Phe Gln Leu Glu Thr Tyr Lys Thr Leu Leu Leu Pro Ile210 215 220tat gcg caa gct gct aat ttt cat tta aat tta tta caa caa ggt gct 720Tyr Ala Gln Ala Ala Asn Phe His Leu Asn Leu Leu Gln Gln Gly Ala225 230 235 240
gaa ttg gct gat gaa tgg aat gca gat ata cat cct tca caa att gaa 768Glu Leu Ala Asp Glu Trp Asn Ala Asp Ile His Pro Ser Gln Ile Glu245 250 255cct aat gct gga aca tca gat gac tat tat aaa ctt tta aaa gaa aat 816Pro Asn Ala Gly Thr Ser Asp Asp Tyr Tyr Lys Leu Leu Lys Glu Asn260 265 270ata cct aaa tat agt aac tat tgt gca aat acc tat aga gaa gga cta 864Ile Pro Lys Tyr Ser Asn Tyr Cys Ala Asn Thr Tyr Arg Glu Gly Leu275 280 285aat aaa ctt cga aac gaa cct aat atg aga tgg agt ata ttt aat gat 912Asn Lys Leu Arg Asn Glu Pro Asn Met Arg Trp Ser Ile Phe Asn Asp290 295 300tat cga aga tat atg act att act gta tta gat act atc gct caa ttt 960Tyr Arg Arg Tyr Met Thr Ile Thr Val Leu Asp Thr Ile Ala Gln Phe305 310 315 320tct ttt tat gat ata aag aga tac aaa gat tca ata gga aga ata ggt 1008Ser Phe Tyr Asp Ile Lys Arg Tyr Lys Asp Ser Ile Gly Arg Ile Gly325 330 335ggc att aaa act gaa ctt aca aga gaa att tat aca act gaa ata aat 1056Gly Ile Lys Thr Glu Leu Thr Arg Glu Ile Tyr Thr Thr Glu Ile Asn340 345 350ttt gac cgt ctt act tac ctt gaa att caa ccc aat ctc gct ata atg 1104Phe Asp Arg Leu Thr Tyr Leu Glu Ile Gln Pro Asn Leu Ala Ile Met355 360 365gaa tat aat tta aca cgt tca ggg ctt aga tta ttt tca ttt tta gat 1152Glu Tyr Asn Leu Thr Arg Ser Gly Leu Arg Leu Phe Ser Phe Leu Asp370 375 380gaa ctt ata ttt tat aca aaa aat gaa acg tac ggg aat cgt tta gtt 1200Glu Leu Ile Phe Tyr Thr Lys Asn Glu Thr Tyr Gly Asn Arg Leu Val385 390 395 400ggt att gcg aat cgt aat aga tct act tat gct acg aca gga act gaa 1248Gly Ile Ala Asn Arg Asn Arg Ser Thr Tyr Ala Thr Thr Gly Thr Glu405 410 415att ata tat gga gaa aga aca ggt cca ccc aca aca aaa act tta ata 1296Ile Ile Tyr Gly Glu Arg Thr Gly Pro Pro Thr Thr Lys Thr Leu Ile420 425 430cca ttt gaa tcc tat aaa gtt tca att gta act gat aga caa gta act 1344Pro Phe Glu Ser Tyr Lys Val Ser Ile Val Thr Asp Arg Gln Val Thr435 440 445cct act tcc cct ttt cct aac ata tac ttt aca att aat caa att gaa 1392Pro Thr Ser Pro Phe Pro Asn Ile Tyr Phe Thr Ile Asn Gln Ile Glu450 455 460ctt tat tta aat aat tca cct agt aat aaa tta aca tat tca gct ggg 1440Leu Tyr Leu Asn Asn Ser Pro Ser Asn Lys Leu Thr Tyr Ser Ala Gly465 470 475 480ggg aat tta tct aat gat aaa aaa aca act gat ttt caa ttt cct gta 1488Gly Asn Leu Ser Asn Asp Lys Lys Thr Thr Asp Phe Gln Phe Pro Val485 490 495aaa aaa gac tgt aaa cca att att aat cca aat tgt tta cca agc tat 1536Lys Lys Asp Cys Lys Pro Ile Ile Asn Pro Asn Cys Leu Pro Ser Tyr500 505 510
aat agt tat agt cat att tta tcc cag ttt tct tta ttt aat tat tcc 1584Asn Ser Tyr Ser His Ile Leu Ser Gln Phe Ser Leu Phe Asn Tyr Ser515 520 525tat aaa att gga tta gcg cta aat ata tta tat aca ggt gca tta gga 1632Tyr Lys Ile Gly Leu Ala Leu Asn Ile Leu Tyr Thr Gly Ala Leu Gly530 535 540tgg aca cac agt agt gtt aat aga aat aat gca ata tca gat aaa ata 1680Trp Thr His Ser Ser Val Asn Arg Asn Asn Ala Ile Ser Asp Lys Ile545 550 555 560att aca atg atc cca gca atc aaa ggt aac agt ctt gat aca aac tct 1728Ile Thr Met Ile Pro Ala Ile Lys Gly Asn Ser Leu Asp Thr Asn Ser565 570 575aag gta att gaa gga cct ggt cat aca gga gga aac ttg gtt tat tta 1776Lys Val Ile Glu Gly Pro Gly His Thr Gly Gly Asn Leu Val Tyr Leu580 585 590caa agt caa ggg cgt tta gag att aca tgt aga act cct aat tct aca 1824Gln Ser Gln Gly Arg Leu Glu Ile Thr Cys Arg Thr Pro Asn Ser Thr595 600 605caa tct tat tac att aga ctt cga tac gct aca aat ggt gct gga aat 1872Gln Ser Tyr Tyr Ile Arg Leu Arg Tyr Ala Thr Asn Gly Ala Gly Asn610 615 620act ctt cct aat ata tct ctt aca ata cca gga gta ata gga ata cca 1920Thr Leu Pro Asn Ile Ser Leu Thr Ile Pro Gly Val Ile Gly Ile Pro625 630 635 640cct caa cga ctc aac aac act ttt tct ggt aca aat tat aat aat tta 1968Pro Gln Arg Leu Asn Asn Thr Phe Ser Gly Thr Asn Tyr Asn Asn Leu645 650 655caa tac gga gat ttt ggg tat ttc caa ttt cca agt aca gta aca tta 2016Gln Tyr Gly Asp Phe Gly Tyr Phe Gln Phe Pro Ser Thr Val Thr Leu660 665 670cct tta aat cga aac ata cca ttt ata ttt aat cgt gca gat gta tca 2064Pro Leu Asn Arg Asn Ile Pro Phe Ile Phe Asn Arg Ala Asp Val Ser675 680 685aat tca att tta atc att gat aaa att gaa ttt ata cca att act tcc 2112Asn Ser Ile Leu Ile Ile Asp Lys Ile Glu Phe Ile Pro Ile Thr Ser690 695 700tct gta cgc caa aat aga gaa aaa caa aaa tta gaa act atc caa aca 2160Ser Val Arg Gln Asn Arg Glu Lys Gln Lys Leu Glu Thr Ile Gln Thr705 710 715 720aaa ata aat aca ttt ttc aca aat cat aca aaa aat act tta aat ata 2208Lys Ile Asn Thr Phe Phe Thr Asn His Thr Lys Asn Thr Leu Asn Ile725 730 735gaa gcc aca aac tat gat att gat taa 2235Glu Ala Thr Asn Tyr Asp Ile Asp *740<210>9<211>744<212>PRT<213>Bacillus thuringiensis<400>9
Met Asn Gln Asn Asn Asn Asn Glu Tyr Glu Ile Ile Asp Ser Lys Asn1 5 10 15Leu Ser Tyr Pro Ser Asn Arg Asn Ile Asp His Ser Arg Tyr Pro Tyr20 25 30Thr Asn Asn Pro Asn Gln Pro Leu Gln Asn Thr Asn Tyr Lys Glu Trp35 40 45Leu Asn Met Cys Gln Gly Asn Thr Gln Tyr Gly Asp Asn Phe Glu Thr50 55 60Phe Ala Ser Ala Asp Thr Ile Ala Ala Val Ser Ala Gly Thr Ile Val65 70 75 80Ser Gly Thr Leu Leu Ala Gly Ile Gly Gly Leu Thr Ser Ile Ser Gly85 90 95Pro Ile Gly Ile Ile Gly Ala Ile Ile Ile Ser Phe Gly Thr Leu Ile100 105 110Thr Val Phe Trp Pro Ala Gly Glu Gln Asp Lys Thr Val Trp Thr Gln115 120 125Phe Ile Lys Met Gly Glu Ile Phe Val Asp Thr Pro Leu Thr Glu Ser130 135 140Ile Lys Gln Leu Lys Leu Gln Thr Leu Glu Gly Phe Arg Gln Ile Leu145 150 155 160Gln Ser Tyr Asn Thr Ala Leu Asp Asp Trp Arg Lys Leu Lys Arg Leu165 170 175Gln Ala Pro Gly Leu Pro Pro Ser Ser Ala Leu Gln Gln Ala Ala Leu180 185 190Thr Leu Lys Ile Arg Phe Glu Asn Val His Asn Asp Phe Ile Arg Glu195 200 205Ile Pro Gly Phe Gln Leu Glu Thr Tyr Lys Thr Leu Leu Leu Pro Ile210 215 220Tyr Ala Gln Ala Ala Asn Phe His Leu Asn Leu Leu Gln Gln Gly Ala225 230 235 240Glu Leu Ala Asp Glu Trp Asn Ala Asp Ile His Pro Ser Gln Ile Glu245 250 255Pro Asn Ala Gly Thr Ser Asp Asp Tyr Tyr Lys Leu Leu Lys Glu Asn260 265 270Ile Pro Lys Tyr Ser Asn Tyr Cys Ala Asn Thr Tyr Arg Glu Gly Leu275 280 285Asn Lys Leu Arg Asn Glu Pro Asn Met Arg Trp Ser Ile Phe Asn Asp290 295 300Tyr Arg Arg Tyr Met Thr Ile Thr Val Leu Asp Thr Ile Ala Gln Phe305 310 315 320Ser Phe Tyr Asp Ile Lys Arg Tyr Lys Asp Ser Ile Gly Arg Ile Gly325 330 335Gly Ile Lys Thr Glu Leu Thr Arg Glu Ile Tyr Thr Thr Glu Ile Asn340 345 350Phe Asp Arg Leu Thr Tyr Leu Glu Ile Gln Pro Asn Leu Ala Ile Met355 360 365Glu Tyr Asn Leu Thr Arg Ser Gly Leu Arg Leu Phe Ser Phe Leu Asp370 375 380Glu Leu Ile Phe Tyr Thr Lys Asn Glu Thr Tyr Gly Asn Arg Leu Val385 390 395 400Gly Ile Ala Asn Arg Asn Arg Ser Thr Tyr Ala Thr Thr Gly Thr Glu405 410 415Ile Ile Tyr Gly Glu Arg Thr Gly Pro Pro Thr Thr Lys Thr Leu Ile420 425 430Pro Phe Glu Ser Tyr Lys Val Ser Ile Val Thr Asp Arg Gln Val Thr435 440 445Pro Thr Ser Pro Phe Pro Asn Ile Tyr Phe Thr Ile Asn Gln Ile Glu450 455 460Leu Tyr Leu Asn Asn Ser Pro Ser Asn Lys Leu Thr Tyr Ser Ala Gly465 470 475 480Gly Asn Leu Ser Asn Asp Lys Lys Thr Thr Asp Phe Gln Phe Pro Val485 490 495Lys Lys Asp Cys Lys Pro Ile Ile Asn Pro Asn Cys Leu Pro Ser Tyr500 505 510Asn Ser Tyr Ser His Ile Leu Ser Gln Phe Ser Leu Phe Asn Tyr Ser515 520 525Tyr Lys Ile Gly Leu Ala Leu Asn Ile Leu Tyr Thr Gly Ala Leu Gly530 535 540
Trp Thr His Ser Ser Val Asn Arg Asn Asn Ala Ile Ser Asp Lys Ile545 550 555 560Ile Thr Met Ile Pro Ala Ile Lys Gly Asn Ser Leu Asp Thr Asn Ser565 570 575Lys Val Ile Glu Gly Pro Gly His Thr Gly Gly Asn Leu Val Tyr Leu580 585 590Gln Ser Gln Gly Arg Leu Glu Ile Thr Cys Arg Thr Pro Asn Ser Thr595 600 605Gln Ser Tyr Tyr Ile Arg Leu Arg Tyr Ala Thr Asn Gly Ala Gly Asn610 615 620Thr Leu Pro Asn Ile Ser Leu Thr Ile Pro Gly Val Ile Gly Ile Pro625 630 635 640Pro Gln Arg Leu Asn Asn Thr Phe Ser Gly Thr Asn Tyr Asn Asn Leu645 650 655Gln Tyr Gly Asp Phe Gly Tyr Phe Gln Phe Pro Ser Thr Val Thr Leu660 665 670Pro Leu Asn Arg Asn Ile Pro Phe Ile Phe Asn Arg Ala Asp Val Ser675 680 685Asn Ser Ile Leu Ile Ile Asp Lys Ile Glu Phe Ile Pro Ile Thr Ser690 695 700Ser Val Arg Gln Asn Arg Glu Lys Gln Lys Leu Glu Thr Ile Gln Thr705 710 715 720Lys Ile Asn Thr Phe Phe Thr Asn His Thr Lys Asn Thr Leu Asn Ile725 730 735Glu Ala Thr Asn Tyr Asp Ile Asp740<210>10<211>2085<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2085)<400>10atg tgt caa ggg aat aca caa tat ggt gat aat ttc gag aca ttt gct 48Met Cys Gln Gly Asn Thr Gln Tyr Gly Asp Asn Phe Glu Thr Phe Ala1 5 10 15agt gct gat aca att gct gca gtt agt gca ggt act att gta tcc ggt 96Ser Ala Asp Thr Ile Ala Ala Val Ser Ala Gly Thr Ile Val Ser Gly20 25 30act ctg tta gcc ggt ata ggt ggg ctc act tct ata tcc gga ccg ata 144Thr Leu Leu Ala Gly Ile Gly Gly Leu Thr Ser Ile Ser Gly Pro Ile35 40 45gga ata ata ggt gct ata ata ata tct ttt ggt acc cta atc act gtc 192Gly Ile Ile Gly Ala Ile Ile Ile Ser Phe Gly Thr Leu Ile Thr Val50 55 60ttt tgg ccc gcg gga gaa caa gac aaa aca gta tgg aca caa ttt att 240Phe Trp Pro Ala Gly Glu Gln Asp Lys Thr Val Trp Thr Gln Phe Ile65 70 75 80aaa atg gga gaa att ttt gtt gat aca ccg tta aca gaa agc ata aaa 288Lys Met Gly Glu Ile Phe Val Asp Thr Pro Leu Thr Glu Ser Ile Lys85 90 95cag cta aag tta caa act tta gaa gga ttt aga caa ata tta caa agc 336Gln Leu Lys Leu Gln Thr Leu Glu Gly Phe Arg Gln Ile Leu Gln Ser100 105 110tat aat aca gca tta gat gat tgg aga aaa tta aaa aga cta caa gct 384Tyr Asn Thr Ala Leu Asp Asp Trp Arg Lys Leu Lys Arg Leu Gln Ala
115 120 125cct gga tta cca cca tca tca gca tta caa caa gct gcc ttg act ctt 432Pro Gly Leu Pro Pro Ser Ser Ala Leu Gln Gln Ala Ala Leu Thr Leu130 135 140aaa ata cga ttt gag aat gtt cac aat gat ttt att cga gaa ata cct 480Lys Ile Arg Phe Glu Asn Val His Asn Asp Phe Ile Arg Glu Ile Pro145 150 155 160ggt ttc caa ctt gaa act tat aaa acg cta tta cta cct att tat gcg 528Gly Phe Gln Leu Glu Thr Tyr Lys Thr Leu Leu Leu Pro Ile Tyr Ala165 170 175caa gct gct aat ttt cat tta aat tta tta caa caa ggt gct gaa ttg 576Gln Ala Ala Asn Phe His Leu Asn Leu Leu Gln Gln Gly Ala Glu Leu180 185 190gct gat gaa tgg aat gca gat ata cat cct tca caa att gaa cct aat 624Ala Asp Glu Trp Asn Ala Asp Ile His Pro Ser Gln Ile Glu Pro Asn195 200 205gct gga aca tca gat gac tat tat aaa ctt tta aaa gaa aat ata cct 672Ala Gly Thr Ser Asp Asp Tyr Tyr Lys Leu Leu Lys Glu Asn Ile Pro210 215 220aaa tat agt aac tat tgt gca aat acc tat aga gaa gga cta aat aaa 720Lys Tyr Ser Asn Tyr Cys Ala Asn Thr Tyr Arg Glu Gly Leu Asn Lys225 230 235 240ctt cga aac gaa cct aat atg aga tgg agt ata ttt aat gat tat cga 768Leu Arg Asn Glu Pro Asn Met Arg Trp Ser Ile Phe Asn Asp Tyr Arg245 250 255aga tat atg act att act gta tta gat act atc gct caa ttt tct ttt 816Arg Tyr Met Thr Ile Thr Val Leu Asp Thr Ile Ala Gln Phe Ser Phe260 265 270tat gat ata aag aga tac aaa gat tca ata gga aga ata ggt ggc att 864Tyr Asp Ile Lys Arg Tyr Lys Asp Ser Ile Gly Arg Ile Gly Gly Ile275 280 285aaa act gaa ctt aca aga gaa att tat aca act gaa ata aat ttt gac 912Lys Thr Glu Leu Thr Arg Glu Ile Tyr Thr Thr Glu Ile Asn Phe Asp290 295 300cgt ctt act tac ctt gaa att caa ccc aat ctc gct ata atg gaa tat 960Arg Leu Thr Tyr Leu Glu Ile Gln Pro Asn Leu Ala Ile Met Glu Tyr305 310 315 320aat tta aca cgt tca ggg ctt aga tta ttt tca ttt tta gat gaa ctt 1008Asn Leu Thr Arg Ser Gly Leu Arg Leu Phe Ser Phe Leu Asp Glu Leu325 330 335ata ttt tat aca aaa aat gaa acg tac ggg aat cgt tta gtt ggt att 1056Ile Phe Tyr Thr Lys Asn Glu Thr Tyr Gly Asn Arg Leu Val Gly Ile340 345 350gcg aat cgt aat aga tct act tat gct acg aca gga act gaa att ata 1104Ala Asn Arg Asn Arg Ser Thr Tyr Ala Thr Thr Gly Thr Glu Ile Ile355 360 365tat gga gaa aga aca ggt cca ccc aca aca aaa act tta ata cca ttt 1152Tyr Gly Glu Arg Thr Gly Pro Pro Thr Thr Lys Thr Leu Ile Pro Phe370 375 380gaa tcc tat aaa gtt tca att gta act gat aga caa gta act cct act 1200Glu Ser Tyr Lys Val Ser Ile Val Thr Asp Arg Gln Val Thr Pro Thr
385 390 395 400tcc cct ttt cct aac ata tac ttt aca att aat caa att gaa ctt tat 1248Ser Pro Phe Pro Asn Ile Tyr Phe Thr Ile Asn Gln Ile Glu Leu Tyr405 410 415tta aat aat tca cct agt aat aaa tta aca tat tca gct ggg ggg aat 1296Leu Asn Asn Ser Pro Ser Asn Lys Leu Thr Tyr Ser Ala Gly Gly Asn420 425 430tta tct aat gat aaa aaa aca act gat ttt caa ttt cct gta aaa aaa 1344Leu Ser Asn Asp Lys Lys Thr Thr Asp Phe Gln Phe Pro Val Lys Lys435 440 445gac tgt aaa cca att att aat cca aat tgt tta cca agc tat aat agt 1392Asp Cys Lys Pro Ile Ile Asn Pro Asn Cys Leu Pro Ser Tyr Asn Ser450 455 460tat agt cat att tta tcc cag ttt tct tta ttt aat tat tcc tat aaa 1440Tyr Ser His Ile Leu Ser Gln Phe Ser Leu Phe Asn Tyr Ser Tyr Lys465 470 475 480att gga tta gcg cta aat ata tta tat aca ggt gca tta gga tgg aca 1488Ile Gly Leu Ala Leu Asn Ile Leu Tyr Thr Gly Ala Leu Gly Trp Thr485 490 495cac agt agt gtt aat aga aat aat gca ata tca gat aaa ata att aca 1536His Ser Ser Val Asn Arg Asn Asn Ala Ile Ser Asp Lys Ile Ile Thr500 505 510atg atc cca gca atc aaa ggt aac agt ctt gat aca aac tct aag gta 1584Met Ile Pro Ala Ile Lys Gly Asn Ser Leu Asp Thr Asn Ser Lys Val515 520 525att gaa gga cct ggt cat aca gga gga aac ttg gtt tat tta caa agt 1632Ile Glu Gly Pro Gly His Thr Gly Gly Asn Leu Val Tyr Leu Gln Ser530 535 540caa ggg cgt tta gag att aca tgt aga act cct aat tct aca caa tct 1680Gln Gly Arg Leu Glu Ile Thr Cys Arg Thr Pro Asn Ser Thr Gln Ser545 550 555 560tat tac att aga ctt cga tac gct aca aat ggt gct gga aat act ctt 1728Tyr Tyr Ile Arg Leu Arg Tyr Ala Thr Asn Gly Ala Gly Asn Thr Leu565 570 575cct aat ata tct ctt aca ata cca gga gta ata gga ata cca cct caa 1776Pro Asn Ile Ser Leu Thr Ile Pro Gly Val Ile Gly Ile Pro Pro Gln580 585 590cga ctc aac aac act ttt tct ggt aca aat tat aat aat tta caa tac 1824Arg Leu Asn Asn Thr Phe Ser Gly Thr Asn Tyr Asn Asn Leu Gln Tyr595 600 605gga gat ttt ggg tat ttc caa ttt cca agt aca gta aca tta cct tta 1872Gly Asp Phe Gly Tyr Phe Gln Phe Pro Ser Thr Val Thr Leu Pro Leu610 615 620aat cga aac ata cca ttt ata ttt aat cgt gca gat gta tca aat tca 1920Asn Arg Asn Ile Pro Phe Ile Phe Asn Arg Ala Asp Val Ser Asn Ser625 630 635 640att tta atc att gat aaa att gaa ttt ata cca att act tcc tct gta 1968Ile Leu Ile Ile Asp Lys Ile Glu Phe Ile Pro Ile Thr Ser Ser Val645 650 655cgc caa aat aga gaa aaa caa aaa tta gaa act atc caa aca aaa ata 2016Arg Gln Asn Arg Glu Lys Gln Lys Leu Glu Thr Ile Gln Thr Lys Ile
660 665 670aat aca ttt ttc aca aat cat aca aaa aat act tta aat ata gaa gcc 2064Asn Thr Phe Phe Thr Asn His Thr Lys Asn Thr Leu Asn Ile Glu Ala675 680 685aca aac tat gat att gat taa 2085Thr Asn Tyr Asp Ile Asp *690<210>11<211>694<212>PRT<213>Bacillus thuringiensis<400>11Met Cys Gln Gly Asn Thr Gln Tyr Gly Asp Asn Phe Glu Thr Phe Ala1 5 10 15Ser Ala Asp Thr Ile Ala Ala Val Ser Ala Gly Thr Ile Val Ser Gly20 25 30Thr Leu Leu Ala Gly Ile Gly Gly Leu Thr Ser Ile Ser Gly Pro Ile35 40 45Gly Ile Ile Gly Ala Ile Ile Ile Ser Phe Gly Thr Leu Ile Thr Val50 55 60Phe Trp Pro Ala Gly Glu Gln Asp Lys Thr Val Trp Thr Gln Phe Ile65 70 75 80Lys Met Gly Glu Ile Phe Val Asp Thr Pro Leu Thr Glu Ser Ile Lys85 90 95Gln Leu Lys Leu Gln Thr Leu Glu Gly Phe Arg Gln Ile Leu Gln Ser100 105 110Tyr Asn Thr Ala Leu Asp Asp Trp Arg Lys Leu Lys Arg Leu Gln Ala115 120 125Pro Gly Leu Pro Pro Ser Ser Ala Leu Gln Gln Ala Ala Leu Thr Leu130 135 140Lys Ile Arg Phe Glu Asn Val His Asn Asp Phe Ile Arg Glu Ile Pro145 150 155 160Gly Phe Gln Leu Glu Thr Tyr Lys Thr Leu Leu Leu Pro Ile Tyr Ala165 170 175Gln Ala Ala Asn Phe His Leu Asn Leu Leu Gln Gln Gly Ala Glu Leu180 185 190Ala Asp Glu Trp Asn Ala Asp Ile His Pro Ser Gln Ile Glu Pro Asn195 200 205Ala Gly Thr Ser Asp Asp Tyr Tyr Lys Leu Leu Lys Glu Asn Ile Pro210 215 220Lys Tyr Ser Asn Tyr Cys Ala Asn Thr Tyr Arg Glu Gly Leu Asn Lys225 230 235 240Leu Arg Asn Glu Pro Asn Met Arg Trp Ser Ile Phe Asn Asp Tyr Arg245 250 255Arg Tyr Met Thr Ile Thr Val Leu Asp Thr Ile Ala Gln Phe Ser Phe260 265 270Tyr Asp Ile Lys Arg Tyr Lys Asp Ser Ile Gly Arg Ile Gly Gly Ile275 280 285Lys Thr Glu Leu Thr Arg Glu Ile Tyr Thr Thr Glu Ile Asn Phe Asp290 295 300Arg Leu Thr Tyr Leu Glu Ile Gln Pro Asn Leu Ala Ile Met Glu Tyr305 310 315 320Asn Leu Thr Arg Ser Gly Leu Arg Leu Phe Ser Phe Leu Asp Glu Leu325 330 335Ile Phe Tyr Thr Lys Asn Glu Thr Tyr Gly Asn Arg Leu Val Gly Ile340 345 350Ala Asn Arg Asn Arg Ser Thr Tyr Ala Thr Thr Gly Thr Glu Ile Ile355 360 365Tyr Gly Glu Arg Thr Gly Pro Pro Thr Thr Lys Thr Leu Ile Pro Phe370 375 380Glu Ser Tyr Lys Val Ser Ile Val Thr Asp Arg Gln Val Thr Pro Thr385 390 395 400Ser Pro Phe Pro Asn Ile Tyr Phe Thr Ile Asn Gln Ile Glu Leu Tyr
405 410 415Leu Asn Asn Ser Pro Ser Asn Lys Leu Thr Tyr Ser Ala Gly Gly Asn420 425 430Leu Ser Asn Asp Lys Lys Thr Thr Asp Phe Gln Phe Pro Val Lys Lys435 440 445Asp Cys Lys Pro Ile Ile Asn Pro Asn Cys Leu Pro Ser Tyr Asn Ser450 455 460Tyr Ser His Ile Leu Ser Gln Phe Ser Leu Phe Asn Tyr Ser Tyr Lys465 470 475 480Ile Gly Leu Ala Leu Asn Ile Leu Tyr Thr Gly Ala Leu Gly Trp Thr485 490 495His Ser Ser Val Asn Arg Asn Asn Ala Ile Ser Asp Lys Ile Ile Thr500 505 510Met Ile Pro Ala Ile Lys Gly Asn Ser Leu Asp Thr Asn Ser Lys Val515 520 525Ile Glu Gly Pro Gly His Thr Gly Gly Asn Leu Val Tyr Leu Gln Ser530 535 540Gln Gly Arg Leu Glu Ile Thr Cys Arg Thr Pro Asn Ser Thr Gln Ser545 550 555 560Tyr Tyr Ile Arg Leu Arg Tyr Ala Thr Asn Gly Ala Gly Asn Thr Leu565 570 575Pro Asn Ile Ser Leu Thr Ile Pro Gly Val Ile Gly Ile Pro Pro Gln580 585 590Arg Leu Asn Asn Thr Phe Ser Gly Thr Asn Tyr Asn Asn Leu Gln Tyr595 600 605Gly Asp Phe Gly Tyr Phe Gln Phe Pro Ser Thr Val Thr Leu Pro Leu610 615 620Asn Arg Asn Ile Pro Phe Ile Phe Asn Arg Ala Asp Val Ser Asn Ser625 630 635 640Ile Leu Ile Ile Asp Lys Ile Glu Phe Ile Pro Ile Thr Ser Ser Val645 650 655Arg Gln Asn Arg Glu Lys Gln Lys Leu Glu Thr Ile Gln Thr Lys Ile660 665 670Asn Thr Phe Phe Thr Asn His Thr Lys Asn Thr Leu Asn Ile Glu Ala675 680 685Thr Asn Tyr Asp Ile Asp690<210>12<211>5980<212>DNA<213>Bacillus thruingiensis<400>12tacatgcaat acataaagag aaggtttaaa aaatcaatac ctcaccaaaa ataatgggtt 60tatttgtaga aacattgtta caggaataca ttggggtact acgaatatat agaaagacac 120ctaacatata tttattaggt gtcttaaaaa taaggactat ataaggagtg aaaaagatga 180gtccatatca aaataaaaat gaatatgaaa tattggaatc ctcatcgaat aacacaaata 240cgccaaacag atatcctttt gcaaataatc gggatatgtc tactatgtct tggaatgatt 300gtcagggaat ctcatgggat gaaatttggg aatcagtcga aacgataaca agtattggga 360taaatcttat agagtttgtg atagaaccta gtttgggtgg aattaataca ctattatcaa 420taataggaaa actaattccg actaatcgtc aaactgtgtc agcactttct atatgtgatt 480tattatctat aattcgtaaa gaggtagccg atagtgtttt aagtgatgcg attgcagatt 540ttgacggtaa attgaaaaat tatagagagt attatctttc ttatcttggg gcttggctta 600aagacggtaa accacttcaa aagacaaata attctgatat cggacaatta gtttattatt 660ttaaactttc agaaagagat ttcaatgaaa ttctaggagg gtcattgtca agaaacaatg 720ctcaagtatt gttattacct acttttgcac aagctgcaaa tgtgcagtta ttactattaa 780gggatgcagt tcaatataaa gcacaatggt tcccattttt gagtgcagag aatgtaagat 840cggaattaat atcacctaac agtggttgtg attttaccgg tgattactat gagcgattaa 900aatgcaaaac ggcagagtat accaattatt gtttatattg gtatcaggta ggtttaaatc 960agataaaaca gggggggaca ggtgctgaca cttggtcgaa atttaataaa tttcgtagag 1020aaatgacgtt ggcggtattg gatattatcg ctatatttcc aacttatgat tttgagaaat 1080atccattgcc aacacatgta gagttgacta gggaaattta tacagatgca gtgggatatt 1140catcgggaac ttatagttgg ttacggaatt ggcctaatac ttttaatggg ttagaggcta 1200atggaacacg gggacctggt ttagttactt ggcttagcaa aataggtata tataatgagt 1260atgtttcgag atattttgcc ggctgggtag gaactcgtca ttatgaagac tacacaaagg 1320gtaacggtat ttttcaacgt atgtctggaa ctacgagtaa tgatctacgt aatattgatt 1380
ttcagaatgc cgatgtatat aaaattactt cattagctat catgaaccta gtaggagaga 1440ctaccgctag accagagtat cgtgtttcaa aggcagattt tcgtagggta gggggacctg 1500atttaaatta tgatgcaggt aataatgggc taagcaggat gacaattgaa tctacgttcc 1560cacttgtatt gcactctaat ggtgttagag gaccctctca tagattatca aatgcggcat 1620gtgttgtata tggaaactcc agagttaacg tatatggttg gacacataca agtttaaaac 1680gtgaaaatat aattgaagcc aatcaaatta cacaaatacc ggcggtgaag agttattacc 1740ttcaaaatta tcttgctaat gcctatacct atgtaataaa aggcactcat acaggtgggg 1800atttaatccg ttttttaaga acaaaatcag agtataacgc agtttatgca ggtggcggaa 1860ttagattgat tattaataac aaaactgcag gacaaagtta ccgtattcgt tttcgttatg 1920ctgcagataa agctgctttc tttagtgtat atctttatcc aggaggttgg ggttcaaatc 1980gttttgtatc gcttgaaaaa tcttactctg gaaattatga cgatttaaaa tatagtgatt 2040ttaaattcgc tgaaattatc acacctccat tacctagttc aaacattcag atggatgtgg 2100agatgcaagc gaatagtttt caatcagatg taaacgtggt tctcgacaaa attgaattcc 2160tcccaagtaa tacaacaact ttagaatatg agggagaacg ggacctagaa aaaacaaaga 2220acgcggtgaa cgatctgttt accaattaaa acaaaataat ttactagaat aggtggtatt 2280gctgtttaac aaataagcga aaaaggttgt gagtcctatg tttacaagta gtacgaaaaa 2340tacgttaaaa atagaaacga cagattatga aatagatcaa gcggccattt ctatagaatg 2400tatgtcagat gaacaaaatc ctcaggaaaa aataatgtta tgggatgaaa taaaactggc 2460aaaacaactt agtcagtctc gtaatctact ccaaaatgga gacttttctg ggaatgattg 2520gacattcggt aatgatatta tcataggatc caataatcct atttttaaag gaaaatttct 2580acagatgcgt ggagcacgag acatatatgg aactctattt ccaacctata tctgtcaaaa 2640aatagatgag tctaaattaa aaccatatac acgttatcga gtaagagggt ttgtgggaag 2700tagtaaagat ttgaaattaa tggtaacacg ttacgggaaa gaaattgatg ctatcatgaa 2760tgttccaaat gatttggcct atatgcagcc taatccttca tgtggagatt atcgctgtga 2820atcatcgtct cagtatgtga gccaagggta tcctacacca acagatggat atgctcccga 2880tatgtatgca tgcccgcaaa atatagatag aaagcatgtg aagtgtcacg atcgtcatcc 2940atttgatttt catattgaca ccggagaagt agatacaaat acaaatgtag gtattgatgt 3000cttattaaaa atttctaatc cagatggata cgctacagta gggaatctag aagtcattga 3060agaaggacca ctaacaggtg aagcattggc acatgtgaaa caaaaggaaa agaaatggaa 3120acaacacatg gagaaaaaac gttgggaaac acaacaagcc tatgatccag caaaacaggc 3180tgtagatgca ttatttacaa atgaacaaga gttacactat catattactt tagatcatat 3240tcaaaacgct gatcgactgg tacagtcgat tccctatgta taccataatt ggttaccgaa 3300tgctccaggt atgaactatg atgtatatca agagttaaac gcacgtatca tgcaaggtta 3360taatttatat gatgcacgaa atgtcataac aaatggtgac tttacacaag gattacaggg 3420atggcacgca acaggaaatg ccgcggtaca acaaatggat ggagcttcag tattagttct 3480atcaaattgg agcgcggggg tatctcaaaa cttgcatgct caagatcatc atggatatgt 3540gttacgtgtg attgccaaaa aagaaggacc tggaaaaggg tatgtaacga tgatggattg 3600taatggaaag caggaaacac ttaagttcac ttcttgcgaa gaaggatata tgacaaaaac 3660agtagaggta ttcccagaaa gtgatcgtgt acggattgaa ataggagaaa ccgaaggtac 3720attttatata gatagcatcg agttgctttg tatgcaagga tatgataaca ataataacct 3780gcacacgggt aatatgtatg agcaaagtta taatggaaat tataatcaaa atactagcga 3840tgtgtattac caagggtata caaacaacta taaccaagac tctagtaata tgtataatca 3900aaattatact aacaatgatg acctgcattc cggttgcaca tgtaaccaag ggcataactc 3960tggctgtaca tgtaatcaag gatataaccg ttaacgattc taaataagaa tcaacatcat 4020tgcgaaaaat aaaaacctac tcacaaaatc tattgcatat cataacataa gctttacaaa 4080taacggacat attctagaag aggtctcctt aattctaaaa taaggagatt tttttcgttt 4140ccccaatatt gattaatgaa aatactcctt tacagaaaga tttaggctga ttggtgtttt 4200gtgcaagcag caattcctga aaaatgcaga aaaaattcca gaggcatgca gctacttttt 4260taaatgttaa ttttggtatt tccagaagat aaagcatttc gtaaacgata actttcacct 4320gtgaatgcaa tgatatgagc atgatggacc aaacggtcca ccaatgcaga agtaagccgg 4380ttatcaccaa atatacgatt ccattgacca aattctaagt ttgaagtaac tatgacactt 4440ttttgttcgt aacagtcggc aataatatga aataggagct ccgctccttg cttttgaaat 4500ggtacataac ctagttcatc taaaattaat aaatcacagg cttcaatttg cttcttgatg 4560cgggttaaat tcccttgttt taaagcctct tctaatagac ctactaaatc tgctactcgg 4620aaaaacttca cttgatatcc ctgttcacaa gccttcaaac ctaatgctgt agctaaatgt 4680gttttcccag ttccaggaga tccaagtaat aaaagatttt gtttcccttc aaaaaatcgc 4740aattcacaaa tgtgttcttg attcgttgta gatggcagat gaatttgttc actccattca 4800taattcttga gccattttaa ctctcgaaat ttcgctttct tgattaagtt tgctactttt 4860ttcgtttgac gagattcaat ctctatattg aaaatatcac gcagaaattg ttctttggtt 4920tcaaaaggta tttcatcata atggtctata atatagctta aatgcagtga tttacaggcg 4980tcttgcaggt ttcgttgtat cataatgaga atgaacccct ttctaaagtt ggacaaagct 5040ttcgatcata ttgctgaaga tctgtttcgt aatctattaa aacaggtggc gtatggtttt 5100cttgccattt ttgtggatac gtcacttgtt gagtctgcaa tatgatatcg agttcgtgag 5160gagctttttt caaccattct tcattctcta acagtaagtg tagctgctgt aatgagtatt 5220tttgaattaa atatttgaat cctgttaaac aagcttttct ttcatcctta tgaaagagca 5280gataatcttg aacccgctga ggtaaatatt tgaaaaatct agaataacga acaacgcgtg 5340gtttcttttc ccaatcttca aaaatctctt cccatagaat cggcctactc gtgtgcatat 5400acggtctgta ttctgtaaaa atgttctcgc cttctgcggt ataacatgta aattgatccc 5460
attctttttt aatgataatc acttgtttta cgtaaaacaa aacgttcttg atccactgtg 5520atttctccat atttatttac cgttgttgtg tccattgaga agaccgttag gtcttccaaa 5580ggtaatggtt tcaatgcagc cttatcatca ttccaaaggt cttcaatcat gacttccttc 5640tcataatgta ggcgtttctg atcttctatt gcttgtactt ctaaccattg tgcgagttga 5700gaaaaactct ccataatcgg agcggttgta aaccaattgt ttcgtgtata actgactttc 5760cgttcaacgt tccccttttc atgcccgcta taaggattac aaggttgcac ttcaaagtta 5820taatgcattt gaaattgtaa aaatgcatcc gtataagttc gattatcacc ttttcctact 5880gtgactacag cggcagataa gttatcaatt cgaagatgcg ttggtacacc accagcttga 5940tgaaataatc gttttaagcc ttctaaaaag ccttctgtat 5980<210>13<211>2082<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2082)<400>13gtg aaa aag atg agt cca tat caa aat aaa aat gaa tat gaa ata ttg 48Met Lys Lys Met Ser Pro Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Leu1 5 10 15gaa tcc tca tcg aat aac aca aat acg cca aac aga tat cct ttt gca 96Glu Ser Ser Ser Asn Asn Thr Asn Thr Pro Asn Arg Tyr Pro Phe Ala20 25 30aat aat cgg gat atg tct act atg tct tgg aat gat tgt cag gga atc 144Asn Asn Arg Asp Met Ser Thr Met Ser Trp Asn Asp Cys Gln Gly Ile35 40 45tca tgg gat gaa att tgg gaa tca gtc gaa acg ata aca agt att ggg 192Ser Trp Asp Glu Ile Trp Glu Ser Val Glu Thr Ile Thr Ser Ile Gly50 55 60ata aat ctt ata gag ttt gtg ata gaa cct agt ttg ggt gga att aat 240Ile Asn Leu Ile Glu Phe Val Ile Glu Pro Ser Leu Gly Gly Ile Asn65 70 75 80aca cta tta tca ata ata gga aaa cta att ccg act aat cgt caa act 288Thr Leu Leu Ser Ile Ile Gly Lys Leu Ile Pro Thr Asn Arg Gln Thr85 90 95gtg tca gca ctt tct ata tgt gat tta tta tct ata att cgt aaa gag 336Val Ser Ala Leu Ser Ile Cys Asp Leu Leu Ser Ile Ile Arg Lys Glu100 105 110gta gcc gat agt gtt tta agt gat gcg att gca gat ttt gac ggt aaa 384Val Ala Asp Ser Val Leu Ser Asp Ala Ile Ala Asp Phe Asp Gly Lys115 120 125ttg aaa aat tat aga gag tat tat ctt tct tat ctt ggg gct tgg ctt 432Leu Lys Asn Tyr Arg Glu Tyr Tyr Leu Ser Tyr Leu Gly Ala Trp Leu130 135 140aaa gac ggt aaa cca ctt caa aag aca aat aat tct gat atc gga caa 480Lys Asp Gly Lys Pro Leu Gln Lys Thr Asn Asn Ser Asp Ile Gly Gln145 150 155 160tta gtt tat tat ttt aaa ctt tca gaa aga gat ttc aat gaa att cta 528Leu Val Tyr Tyr Phe Lys Leu Ser Glu Arg Asp Phe Asn Glu Ile Leu165 170 175gga ggg tca ttg tca aga aac aat gct caa gta ttg tta tta cct act 576Gly Gly Ser Leu Ser Arg Asn Asn Ala Gln Val Leu Leu Leu Pro Thr
180 185 190ttt gca caa gct gca aat gtg cag tta tta cta tta agg gat gca gtt 624Phe Ala Gln Ala Ala Asn Val Gln Leu Leu Leu Leu Arg Asp Ala Val195 200 205caa tat aaa gca caa tgg ttc cca ttt ttg agt gca gag aat gta aga 672Gln Tyr Lys Ala Gln Trp Phe Pro Phe Leu Ser Ala Glu Asn Val Arg210 215 220tcg gaa tta ata tca cct aac agt ggt tgt gat ttt acc ggt gat tac 720Ser Glu Leu Ile Ser Pro Asn Ser Gly Cys Asp Phe Thr Gly Asp Tyr225 230 235 240tat gag cga tta aaa tgc aaa acg gca gag tat acc aat tat tgt tta 768Tyr Glu Arg Leu Lys Cys Lys Thr Ala Glu Tyr Thr Asn Tyr Cys Leu245 250 255tat tgg tat cag gta ggt tta aat cag ata aaa cag ggg ggg aca ggt 816Tyr Trp Tyr Gln Val Gly Leu Asn Gln Ile Lys Gln Gly Gly Thr Gly260 265 270gct gac act tgg tcg aaa ttt aat aaa ttt cgt aga gaa atg acg ttg 864Ala Asp Thr Trp Ser Lys Phe Asn Lys Phe Arg Arg Glu Met Thr Leu275 280 285gcg gta ttg gat att atc gct ata ttt cca act tat gat ttt gag aaa 912Ala Val Leu Asp Ile Ile Ala Ile Phe Pro Thr Tyr Asp Phe Glu Lys290 295 300tat cca ttg cca aca cat gta gag ttg act agg gaa att tat aca gat 960Tyr Pro Leu Pro Thr His Val Glu Leu Thr Arg Glu Ile Tyr Thr Asp305 310 315 320gca gtg gga tat tca tcg gga act tat agt tgg tta cgg aat tgg cct 1008Ala Val Gly Tyr Ser Ser Gly Thr Tyr Ser Trp Leu Arg Asn Trp Pro325 330 335aat act ttt aat ggg tta gag gct aat gga aca cgg gga cct ggt tta 1056Asn Thr Phe Asn Gly Leu Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu340 345 350gtt act tgg ctt agc aaa ata ggt ata tat aat gag tat gtt tcg aga 1104Val Thr Trp Leu Ser Lys Ile Gly Ile Tyr Asn Glu Tyr Val Ser Arg355 360 365tat ttt gcc ggc tgg gta gga act cgt cat tat gaa gac tac aca aag 1152Tyr Phe Ala Gly Trp Val Gly Thr Arg His Tyr Glu Asp Tyr Thr Lys370 375 380ggt aac ggt att ttt caa cgt atg tct gga act acg agt aat gat cta 1200Gly Asn Gly Ile Phe Gln Arg Met Ser Gly Thr Thr Ser Asn Asp Leu385 390 395 400cgt aat att gat ttt cag aat gcc gat gta tat aaa att act tca tta 1248Arg Asn Ile Asp Phe Gln Asn Ala Asp Val Tyr Lys Ile Thr Ser Leu405 410 415gct atc atg aac cta gta gga gag act acc gct aga cca gag tat cgt 1296Ala Ile Met Asn Leu Val Gly Glu Thr Thr Ala Arg Pro Glu Tyr Arg420 425 430gtt tca aag gca gat ttt cgt agg gta ggg gga cct gat tta aat tat 1344Val Ser Lys Ala Asp Phe Arg Arg Val Gly Gly Pro Asp Leu Asn Tyr435 440 445gat gca ggt aat aat ggg cta agc agg atg aca att gaa tct acg ttc 1392Asp Ala Gly Asn Asn Gly Leu Ser Arg Met Thr Ile Glu Ser Thr Phe
450 455 460cca ctt gta ttg cac tct aat ggt gtt aga gga ccc tct cat aga tta 1440Pro Leu Val Leu His Ser Asn Gly Val Arg Gly Pro Ser His Arg Leu465 470 475 480tca aat gcg gca tgt gtt gta tat gga aac tcc aga gtt aac gta tat 1488Ser Asn Ala Ala Cys Val Val Tyr Gly Asn Ser Arg Val Asn Val Tyr485 490 495ggt tgg aca cat aca agt tta aaa cgt gaa aat ata att gaa gcc aat 1536Gly Trp Thr His Thr Ser Leu Lys Arg Glu Asn Ile Ile Glu Ala Asn500 505 5l0caa att aca caa ata ccg gcg gtg aag agt tat tac ctt caa aat tat 1584Gln Ile Thr Gln Ile Pro Ala Val Lys Ser Tyr Tyr Leu Gln Asn Tyr515 520 525ctt gct aat gcc tat acc tat gta ata aaa ggc act cat aca ggt ggg 1632Leu Ala Asn Ala Tyr Thr Tyr Val Ile Lys Gly Thr His Thr Gly Gly530 535 540gat tta atc cgt ttt tta aga aca aaa tca gag tat aac gca gtt tat 1680Asp Leu Ile Arg Phe Leu Arg Thr Lys Ser Glu Tyr Asn Ala Val Tyr545 550 555 560gca ggt ggc gga att aga ttg att att aat aac aaa act gca gga caa 1728Ala Gly Gly Gly Ile Arg Leu Ile Ile Asn Asn Lys Thr Ala Gly Gln565 570 575agt tac cgt att cgt ttt cgt tat gct gca gat aaa gct gct ttc ttt 1776Ser Tyr Arg Ile Arg Phe Arg Tyr Ala Ala Asp Lys Ala Ala Phe Phe580 585 590agt gta tat ctt tat cca gga ggt tgg ggt tca aat cgt ttt gta tcg 1824Ser Val Tyr Leu Tyr Pro Gly Gly Trp Gly Ser Asn Arg Phe Val Ser595 600 605ctt gaa aaa tct tac tct gga aat tat gac gat tta aaa tat agt gat 1872Leu Glu Lys Ser Tyr Ser Gly Asn Tyr Asp Asp Leu Lys Tyr Ser Asp610 615 620ttt aaa ttc gct gaa att atc aca cct cca tta cct agt tca aac att 1920Phe Lys Phe Ala Glu Ile Ile Thr Pro Pro Leu Pro Ser Ser Asn Ile625 630 635 640cag atg gat gtg gag atg caa gcg aat agt ttt caa tca gat gta aac 1968Gln Met Asp Val Glu Met Gln Ala Asn Ser Phe Gln Ser Asp Val Asn645 650 655gtg gtt ctc gac aaa att gaa ttc ctc cca agt aat aca aca act tta 2016Val Val Leu Asp Lys Ile Glu Phe Leu Pro Ser Asn Thr Thr Thr Leu660 665 670gaa tat gag gga gaa cgg gac cta gaa aaa aca aag aac gcg gtg aac 2064Glu Tyr Glu Gly Glu Arg Asp Leu Glu Lys Thr Lys Asn Ala Val Asn675 680 685gat ctg ttt acc aat taa 2082Asp Leu Phe Thr Asn *690<210>14<211>693<212>PRT<213>Bacillus thuringiensis
<400>14Met Lys Lys Met Ser Pro Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Leu1 5 10 15Glu Ser Ser Ser Asn Asn Thr Asn Thr Pro Asn Arg Tyr Pro Phe Ala20 25 30Asn Asn Arg Asp Met Ser Thr Met Ser Trp Asn Asp Cys Gln Gly Ile35 40 45Ser Trp Asp Glu Ile Trp Glu Ser Val Glu Thr Ile Thr Ser Ile Gly50 55 60Ile Asn Leu Ile Glu Phe Val Ile Glu Pro Ser Leu Gly Gly Ile Asn65 70 75 80Thr Leu Leu Ser Ile Ile Gly Lys Leu Ile Pro Thr Asn Arg Gln Thr85 90 95Val Ser Ala Leu Ser Ile Cys Asp Leu Leu Ser Ile Ile Arg Lys Glu100 105 110Val Ala Asp Ser Val Leu Ser Asp Ala Ile Ala Asp Phe Asp Gly Lys115 120 125Leu Lys Asn Tyr Arg Glu Tyr Tyr Leu Ser Tyr Leu Gly Ala Trp Leu130 135 140Lys Asp Gly Lys Pro Leu Gln Lys Thr Asn Asn Ser Asp Ile Gly Gln145 150 155 160Leu Val Tyr Tyr Phe Lys Leu Ser Glu Arg Asp Phe Asn Glu Ile Leu165 170 175Gly Gly Ser Leu Ser Arg Asn Asn Ala Gln Val Leu Leu Leu Pro Thr180 185 190Phe Ala Gln Ala Ala Asn Val Gln Leu Leu Leu Leu Arg Asp Ala Val195 200 205Gln Tyr Lys Ala Gln Trp Phe Pro Phe Leu Ser Ala Glu Asn Val Arg210 215 220Ser Glu Leu Ile Ser Pro Asn Ser Gly Cys Asp Phe Thr Gly Asp Tyr225 230 235 240Tyr Glu Arg Leu Lys Cys Lys Thr Ala Glu Tyr Thr Asn Tyr Cys Leu245 250 255Tyr Trp Tyr Gln Val Gly Leu Asn Gln Ile Lys Gln Gly Gly Thr Gly260 265 270Ala Asp Thr Trp Ser Lys Phe Asn Lys Phe Arg Arg Glu Met Thr Leu275 280 285Ala Val Leu Asp Ile Ile Ala Ile Phe Pro Thr Tyr Asp Phe Glu Lys290 295 300Tyr Pro Leu Pro Thr His Val Glu Leu Thr Arg Glu Ile Tyr Thr Asp305 310 315 320Ala Val Gly Tyr Ser Ser Gly Thr Tyr Ser Trp Leu Arg Asn Trp Pro325 330 335Asn Thr Phe Asn Gly Leu Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu340 345 350Val Thr Trp Leu Ser Lys Ile Gly Ile Tyr Asn Glu Tyr Val Ser Arg355 360 365Tyr Phe Ala Gly Trp Val Gly Thr Arg His Tyr Glu Asp Tyr Thr Lys370 375 380Gly Asn Gly Ile Phe Gln Arg Met Ser Gly Thr Thr Ser Asn Asp Leu385 390 395 400Arg Asn Ile Asp Phe Gln Asn Ala Asp Val Tyr Lys Ile Thr Ser Leu405 410 415Ala Ile Met Asn Leu Val Gly Glu Thr Thr Ala Arg Pro Glu Tyr Arg420 425 430Val Ser Lys Ala Asp Phe Arg Arg Val Gly Gly Pro Asp Leu Asn Tyr435 440 445Asp Ala Gly Asn Asn Gly Leu Ser Arg Met Thr Ile Glu Ser Thr Phe450 455 460Pro Leu Val Leu His Ser Asn Gly Val Arg Gly Pro Ser His Arg Leu465 470 475 480Ser Asn Ala Ala Cys Val Val Tyr Gly Asn Ser Arg Val Asn Val Tyr485 490 495Gly Trp Thr His Thr Ser Leu Lys Arg Glu Asn Ile Ile Glu Ala Asn500 505 510Gln Ile Thr Gln Ile Pro Ala Val Lys Ser Tyr Tyr Leu Gln Asn Tyr515 520 525Leu Ala Asn Ala Tyr Thr Tyr Val Ile Lys Gly Thr His Thr Gly Gly
530 535 540Asp Leu Ile Arg Phe Leu Arg Thr Lys Ser Glu Tyr Asn Ala Val Tyr545 550 555 560Ala Gly Gly Gly Ile Arg Leu Ile Ile Asn Asn Lys Thr Ala Gly Gln565 570 575Ser Tyr Arg Ile Arg Phe Arg Tyr Ala Ala Asp Lys Ala Ala Phe Phe580 585 590Ser Val Tyr Leu Tyr Pro Gly Gly Trp Gly Ser Asn Arg Phe Val Ser595 600 605Leu Glu Lys Ser Tyr Ser Gly Asn Tyr Asp Asp Leu Lys Tyr Ser Asp610 615 620Phe Lys Phe Ala Glu Ile Ile Thr Pro Pro Leu Pro Ser Ser Asn Ile625 630 635 640Gln Met Asp Val Glu Met Gln Ala Asn Ser Phe Gln Ser Asp Val Asn645 650 655Val Val Leu Asp Lys Ile Glu Phe Leu Pro Ser Asn Thr Thr Thr Leu660 665 670Glu Tyr Glu Gly Glu Arg Asp Leu Glu Lys Thr Lys Asn Ala Val Asn675 680 685Asp Leu Phe Thr Asn690<210>15<211>2073<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2073)<400>15atg agt cca tat caa aat aaa aat gaa tat gaa ata ttg gaa tcc tca 48Met Ser Pro Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Leu Glu Ser Ser1 5 10 15tcg aat aac aca aat acg cca aac aga tat cct ttt gca aat aat cgg 96Ser Asn Asn Thr Asn Thr Pro Asn Arg Tyr Pro Phe Ala Asn Asn Arg20 25 30gat atg tct act atg tct tgg aat gat tgt cag gga atc tca tgg gat 144Asp Met Ser Thr Met Ser Trp Asn Asp Cys Gln Gly Ile Ser Trp Asp35 40 45gaa att tgg gaa tca gtc gaa acg ata aca agt att ggg ata aat ctt 192Glu Ile Trp Glu Ser Val Glu Thr Ile Thr Ser Ile Gly Ile Asn Leu50 55 60ata gag ttt gtg ata gaa cct agt ttg ggt gga att aat aca cta tta 240Ile Glu Phe Val Ile Glu Pro Ser Leu Gly Gly Ile Asn Thr Leu Leu65 70 75 80tca ata ata gga aaa cta att ccg act aat cgt caa act gtg tca gca 288Ser Ile Ile Gly Lys Leu Ile Pro Thr Asn Arg Gln Thr Val Ser Ala85 90 95ctt tct ata tgt gat tta tta tct ata att cgt aaa gag gta gcc gat 336Leu Ser Ile Cys Asp Leu Leu Ser Ile Ile Arg Lys Glu Val Ala Asp100 105 110agt gtt tta agt gat gcg att gca gat ttt gac ggt aaa ttg aaa aat 384Ser Val Leu Ser Asp Ala Ile Ala Asp Phe Asp Gly Lys Leu Lys Asn115 120 125tat aga gag tat tat ctt tct tat ctt ggg gct tgg ctt aaa gac ggt 432Tyr Arg Glu Tyr Tyr Leu Ser Tyr Leu Gly Ala Trp Leu Lys Asp Gly130 135 140
aaa cca ctt caa aag aca aat aat tct gat atc gga caa tta gtt tat 480Lys Pro Leu Gln Lys Thr Asn Asn Ser Asp Ile Gly Gln Leu Val Tyr145 150 155 160tat ttt aaa ctt tca gaa aga gat ttc aat gaa att cta gga ggg tca 528Tyr Phe Lys Leu Ser Glu Arg Asp Phe Asn Glu Ile Leu Gly Gly Ser165 170 175ttg tca aga aac aat gct caa gta ttg tta tta cct act ttt gca caa 576Leu Ser Arg Asn Asn Ala Gln Val Leu Leu Leu Pro Thr Phe Ala Gln180 185 190gct gca aat gtg cag tta tta cta tta agg gat gca gtt caa tat aaa 624Ala Ala Asn Val Gln Leu Leu Leu Leu Arg Asp Ala Val Gln Tyr Lys195 200 205gca caa tgg ttc cca ttt ttg agt gca gag aat gta aga tcg gaa tta 672Ala Gln Trp Phe Pro Phe Leu Ser Ala Glu Asn Val Arg Ser Glu Leu210 215 220ata tca cct aac agt ggt tgt gat ttt acc ggt gat tac tat gag cga 720Ile Ser Pro Asn Ser Gly Cys Asp Phe Thr Gly Asp Tyr Tyr Glu Arg225 230 235 240tta aaa tgc aaa acg gca gag tat acc aat tat tgt tta tat tgg tat 768Leu Lys Cys Lys Thr Ala Glu Tyr Thr Asn Tyr Cys Leu Tyr Trp Tyr245 250 255cag gta ggt tta aat cag ata aaa cag ggg ggg aca ggt gct gac act 816Gln Val Gly Leu Asn Gln Ile Lys Gln Gly Gly Thr Gly Ala Asp Thr260 265 270tgg tcg aaa ttt aat aaa ttt cgt aga gaa atg acg ttg gcg gta ttg 864Trp Ser Lys Phe Asn Lys Phe Arg Arg Glu Met Thr Leu Ala Val Leu275 280 285gat att atc gct ata ttt cca act tat gat ttt gag aaa tat cca ttg 912Asp Ile Ile Ala Ile Phe Pro Thr Tyr Asp Phe Glu Lys Tyr Pro Leu290 295 300cca aca cat gta gag ttg act agg gaa att tat aca gat gca gtg gga 960Pro Thr His Val Glu Leu Thr Arg Glu Ile Tyr Thr Asp Ala Val Gly305 310 315 320tat tca tcg gga act tat agt tgg tta cgg aat tgg cct aat act ttt 1008Tyr Ser Ser Gly Thr Tyr Ser Trp Leu Arg Asn Trp Pro Asn Thr Phe325 330 335aat ggg tta gag gct aat gga aca cgg gga cct ggt tta gtt act tgg 1056Asn Gly Leu Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu Val Thr Trp340 345 350ctt agc aaa ata ggt ata tat aat gag tat gtt tcg aga tat ttt gcc 1104Leu Ser Lys Ile Gly Ile Tyr Asn Glu Tyr Val Ser Arg Tyr Phe Ala355 360 365ggc tgg gta gga act cgt cat tat gaa gac tac aca aag ggt aac ggt 1152Gly Trp Val Gly Thr Arg His Tyr Glu Asp Tyr Thr Lys Gly Asn Gly370 375 380att ttt caa cgt atg tct gga act acg agt aat gat cta cgt aat att 1200Ile Phe Gln Arg Met Ser Gly Thr Thr Ser Asn Asp Leu Arg Asn Ile385 390 395 400gat ttt cag aat gcc gat gta tat aaa att act tca tta gct atc atg 1248Asp Phe Gln Asn Ala Asp Val Tyr Lys Ile Thr Ser Leu Ala Ile Met405 410 415
aac cta gta gga gag act acc gct aga cca gag tat cgt gtt tca aag 1296Asn Leu Val Gly Glu Thr Thr Ala Arg Pro Glu Tyr Arg Val Ser Lys420 425 430gca gat ttt cgt agg gta ggg gga cct gat tta aat tat gat gca ggt 1344Ala Asp Phe Arg Arg Val Gly Gly Pro Asp Leu Asn Tyr Asp Ala Gly435 440 445aat aat ggg cta agc agg atg aca att gaa tct acg ttc cca ctt gta 1392Asn Asn Gly Leu Ser Arg Met Thr Ile Glu Ser Thr Phe Pro Leu Val450 455 460ttg cac tct aat ggt gtt aga gga ccc tct cat aga tta tca aat gcg 1440Leu His Ser Asn Gly Val Arg Gly Pro Ser His Arg Leu Ser Asn Ala465 470 475 480gca tgt gtt gta tat gga aac tcc aga gtt aac gta tat ggt tgg aca 1488Ala Cys Val Val Tyr Gly Asn Ser Arg Val Asn Val Tyr Gly Trp Thr485 490 495cat aca agt tta aaa cgt gaa aat ata att gaa gcc aat caa att aca 1536His Thr Ser Leu Lys Arg Glu Asn Ile Ile Glu Ala Asn Gln Ile Thr500 505 510caa ata ccg gcg gtg aag agt tat tac ctt caa aat tat ctt gct aat 1584Gln Ile Pro Ala Val Lys Ser Tyr Tyr Leu Gln Asn Tyr Leu Ala Asn515 520 525gcc tat acc tat gta ata aaa ggc act cat aca ggt ggg gat tta atc 1632Ala Tyr Thr Tyr Val Ile Lys Gly Thr His Thr Gly Gly Asp Leu Ile530 535 540cgt ttt tta aga aca aaa tca gag tat aac gca gtt tat gca ggt ggc 1680Arg Phe Leu Arg Thr Lys Ser Glu Tyr Asn Ala Val Tyr Ala Gly Gly545 550 555 560gga att aga ttg att att aat aac aaa act gca gga caa agt tac cgt 1728Gly Ile Arg Leu Ile Ile Asn Asn Lys Thr Ala Gly Gln Ser Tyr Arg565 570 575att cgt ttt cgt tat gct gca gat aaa gct gct ttc ttt agt gta tat 1776Ile Arg Phe Arg Tyr Ala Ala Asp Lys Ala Ala Phe Phe Ser Val Tyr580 585 590ctt tat cca gga ggt tgg ggt tca aat cgt ttt gta tcg ctt gaa aaa 1824Leu Tyr Pro Gly Gly Trp Gly Ser Asn Arg Phe Val Ser Leu Glu Lys595 600 605tct tac tct gga aat tat gac gat tta aaa tat agt gat ttt aaa ttc 1872Ser Tyr Ser Gly Asn Tyr Asp Asp Leu Lys Tyr Ser Asp Phe Lys Phe610 615 620gct gaa att atc aca cct cca tta cct agt tca aac att cag atg gat 1920Ala Glu Ile Ile Thr Pro Pro Leu Pro Ser Ser Asn Ile Gln Met Asp625 630 635 640gtg gag atg caa gcg aat agt ttt caa tca gat gta aac gtg gtt ctc 1968Val Glu Met Gln Ala Asn Ser Phe Gln Ser Asp Val Asn Val Val Leu645 650 655gac aaa att gaa ttc ctc cca agt aat aca aca act tta gaa tat gag 2016Asp Lys Ile Glu Phe Leu Pro Ser Asn Thr Thr Thr Leu Glu Tyr Glu660 665 670gga gaa cgg gac cta gaa aaa aca aag aac gcg gtg aac gat ctg ttt 2064Gly Glu Arg Asp Leu Glu Lys Thr Lys Asn Ala Val Asn Asp Leu Phe675 680 685
acc aat taa 2073Thr Asn *690<210>16<211>690<212>PRT<213>Bacillus thuringiensis<400>16Met Ser Pro Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Leu Glu Ser Ser1 5 10 15Ser Asn Asn Thr Asn Thr Pro Asn Arg Tyr Pro Phe Ala Asn Asn Arg20 25 30Asp Met Ser Thr Met Ser Trp Asn Asp Cys Gln Gly Ile Ser Trp Asp35 40 45Glu Ile Trp Glu Ser Val Glu Thr Ile Thr Ser Ile Gly Ile Asn Leu50 55 60Ile Glu Phe Val Ile Glu Pro Ser Leu Gly Gly Ile Asn Thr Leu Leu65 70 75 80Ser Ile Ile Gly Lys Leu Ile Pro Thr Asn Arg Gln Thr Val Ser Ala85 90 95Leu Ser Ile Cys Asp Leu Leu Ser Ile Ile Arg Lys Glu Val Ala Asp100 105 110Ser Val Leu Ser Asp Ala Ile Ala Asp Phe Asp Gly Lys Leu Lys Asn115 120 125Tyr Arg Glu Tyr Tyr Leu Ser Tyr Leu Gly Ala Trp Leu Lys Asp Gly130 135 140Lys Pro Leu Gln Lys Thr Asn Asn Ser Asp Ile Gly Gln Leu Val Tyr145 150 155 160Tyr Phe Lys Leu Ser Glu Arg Asp Phe Asn Glu Ile Leu Gly Gly Ser165 170 175Leu Ser Arg Asn Asn Ala Gln Val Leu Leu Leu Pro Thr Phe Ala Gln180 185 190Ala Ala Asn Val Gln Leu Leu Leu Leu Arg Asp Ala Val Gln Tyr Lys195 200 205Ala Gln Trp Phe Pro Phe Leu Ser Ala Glu Asn Val Arg Ser Glu Leu210 215 220Ile Ser Pro Asn Ser Gly Cys Asp Phe Thr Gly Asp Tyr Tyr Glu Arg225 230 235 240Leu Lys Cys Lys Thr Ala Glu Tyr Thr Asn Tyr Cys Leu Tyr Trp Tyr245 250 255Gln Val Gly Leu Asn Gln Ile Lys Gln Gly Gly Thr Gly Ala Asp Thr260 265 270Trp Ser Lys Phe Asn Lys Phe Arg Arg Glu Met Thr Leu Ala Val Leu275 280 285Asp Ile Ile Ala Ile Phe Pro Thr Tyr Asp Phe Glu Lys Tyr Pro Leu290 295 300Pro Thr His Val Glu Leu Thr Arg Glu Ile Tyr Thr Asp Ala Val Gly305 310 315 320Tyr Ser Ser Gly Thr Tyr Ser Trp Leu Arg Asn Trp Pro Asn Thr Phe325 330 335Asn Gly Leu Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu Val Thr Trp340 345 350Leu Ser Lys Ile Gly Ile Tyr Asn Glu Tyr Val Ser Arg Tyr Phe Ala355 360 365Gly Trp Val Gly Thr Arg His Tyr Glu Asp Tyr Thr Lys Gly Asn Gly370 375 380Ile Phe Gln Arg Met Ser Gly Thr Thr Ser Asn Asp Leu Arg Asn Ile385 390 395 400Asp Phe Gln Asn Ala Asp Val Tyr Lys Ile Thr Ser Leu Ala Ile Met405 410 415Asn Leu Val Gly Glu Thr Thr Ala Arg Pro Glu Tyr Arg Val Ser Lys420 425 430Ala Asp Phe Arg Arg Val Gly Gly Pro Asp Leu Asn Tyr Asp Ala Gly435 440 445
Asn Asn Gly Leu Ser Arg Met Thr Ile Glu Ser Thr Phe Pro Leu Val450 455 460Leu His Ser Asn Gly Val Arg Gly Pro Ser His Arg Leu Ser Asn Ala465 470 475 480Ala Cys Val Val Tyr Gly Asn Ser Arg Val Asn Val Tyr Gly Trp Thr485 490 495His Thr Ser Leu Lys Arg Glu Asn Ile Ile Glu Ala Asn Gln Ile Thr500 505 510Gln Ile Pro Ala Val Lys Ser Tyr Tyr Leu Gln Asn Tyr Leu Ala Asn515 520 525Ala Tyr Thr Tyr Val Ile Lys Gly Thr His Thr Gly Gly Asp Leu Ile530 535 540Arg Phe Leu Arg Thr Lys Ser Glu Tyr Asn Ala Val Tyr Ala Gly Gly545 550 555 560Gly Ile Arg Leu Ile Ile Asn Asn Lys Thr Ala Gly Gln Ser Tyr Arg565 570 575Ile Arg Phe Arg Tyr Ala Ala Asp Lys Ala Ala Phe Phe Ser Val Tyr580 585 590Leu Tyr Pro Gly Gly Trp Gly Ser Asn Arg Phe Val Ser Leu Glu Lys595 600 605Ser Tyr Ser Gly Asn Tyr Asp Asp Leu Lys Tyr Ser Asp Phe Lys Phe610 615 620Ala Glu Ile Ile Thr Pro Pro Leu Pro Ser Ser Asn Ile Gln Met Asp625 630 635 640Val Glu Met Gln Ala Asn Ser Phe Gln Ser Asp Val Asn Val Val Leu645 650 655Asp Lys Ile Glu Phe Leu Pro Ser Asn Thr Thr Thr Leu Glu Tyr Glu660 665 670Gly Glu Arg Asp Leu Glu Lys Thr Lys Asn Ala Val Asn Asp Leu Phe675 680 685Thr Asn690<210>17<211>1686<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(1686)<400>17gtg agt cct atg ttt aca agt agt acg aaa aat acg tta aaa ata gaa 48Met Ser Pro Met Phe Thr Ser Ser Thr Lys Asn Thr Leu Lys Ile Glu1 5 10 15acg aca gat tat gaa ata gat caa gcg gcc att tct ata gaa tgt atg 96Thr Thr Asp Tyr Glu Ile Asp Gln Ala Ala Ile Ser Ile Glu Cys Met20 25 30tca gat gaa caa aat cct cag gaa aaa ata atg tta tgg gat gaa ata 144Ser Asp Glu Gln Asn Pro Gln Glu Lys Ile Met Leu Trp Asp Glu Ile35 40 45aaa ctg gca aaa caa ctt agt cag tct cgt aat cta ctc caa aat gga 192Lys Leu Ala Lys Gln Leu Ser Gln Ser Arg Asn Leu Leu Gln Asn Gly50 55 60gac ttt tct ggg aat gat tgg aca ttc ggt aat gat att atc ata gga 240Asp Phe Ser Gly Asn Asp Trp Thr Phe Gly Asn Asp Ile Ile Ile Gly65 70 75 80tcc aat aat cct att ttt aaa gga aaa ttt cta cag atg cgt gga gca 288Ser Asn Asn Pro Ile Phe Lys Gly Lys Phe Leu Gln Met Arg Gly Ala85 90 95
cga gac ata tat gga act cta ttt cca acc tat atc tgt caa aaa ata 336Arg Asp Ile Tyr Gly Thr Leu Phe Pro Thr Tyr Ile Cys Gln Lys Ile100 105 110gat gag tct aaa tta aaa cca tat aca cgt tat cga gta aga ggg ttt 384Asp Glu Ser Lys Leu Lys Pro Tyr Thr Arg Tyr Arg Val Arg Gly Phe115 120 125gtg gga agt agt aaa gat ttg aaa tta atg gta aca cgt tac ggg aaa 432Val Gly Ser Ser Lys Asp Leu Lys Leu Met Val Thr Arg Tyr Gly Lys130 135 140gaa att gat gct atc atg aat gtt cca aat gat ttg gcc tat atg cag 480Glu Ile Asp Ala Ile Met Asn Val Pro Asn Asp Leu Ala Tyr Met Gln145 150 155 160cct aat cct tca tgt gga gat tat cgc tgt gaa tca tcg tct cag tat 528Pro Asn Pro Ser Cys Gly Asp Tyr Arg Cys Glu Ser Ser Ser Gln Tyr165 170 175gtg agc caa ggg tat cct aca cca aca gat gga tat gct ccc gat atg 576Val Ser Gln Gly Tyr Pro Thr Pro Thr Asp Gly Tyr Ala Pro Asp Met180 185 190tat gca tgc ccg caa aat ata gat aga aag cat gtg aag tgt cac gat 624Tyr Ala Cys Pro Gln Asn Ile Asp Arg Lys His Val Lys Cys His Asp195 200 205cgt cat cca ttt gat ttt cat att gac acc gga gaa gta gat aca aat 672Arg His Pro phe Asp Phe His Ile Asp Thr Gly Glu Val Asp Thr Asn210 215 220aca aat gta ggt att gat gtc tta tta aaa att tct aat cca gat gga 720Thr Asn Val Gly Ile Asp Val Leu Leu Lys Ile Ser Asn Pro Asp Gly225 230 235 240tac gct aca gta ggg aat cta gaa gtc att gaa gaa gga cca cta aca 768Tyr Ala Thr Val Gly Asn Leu Glu Val Ile Glu Glu Gly Pro Leu Thr245 250 255ggt gaa gca ttg gca cat gtg aaa caa aag gaa aag aaa tgg aaa caa 816Gly Glu Ala Leu Ala His Val Lys Gln Lys Glu Lys Lys Trp Lys Gln260 265 270cac atg gag aaa aaa cgt tgg gaa aca caa caa gcc tat gat cca gca 864His Met Glu Lys Lys Arg Trp Glu Thr Gln Gln Ala Tyr Asp Pro Ala275 280 285aaa cag gct gta gat gca tta ttt aca aat gaa caa gag tta cac tat 912Lys Gln Ala Val Asp Ala Leu Phe Thr Asn Glu Gln Glu Leu His Tyr290 295 300cat att act tta gat cat att caa aac gct gat cga ctg gta cag tcg 960His Ile Thr Leu Asp His Ile Gln Asn Ala Asp Arg Leu Val Gln Ser305 310 315 320att ccc tat gta tac cat aat tgg tta ccg aat gct cca ggt atg aac 1008Ile Pro Tyr Val Tyr His Asn Trp Leu Pro Asn Ala Pro Gly Met Asn325 330 335tat gat gta tat caa gag tta aac gca cgt atc atg caa ggt tat aat 1056Tyr Asp Val Tyr Gln Glu Leu Asn Ala Arg Ile Met Gln Gly Tyr Asn340 345 350tta tat gat gca cga aat gtc ata aca aat ggt gac ttt aca caa gga 1104Leu Tyr Asp Ala Arg Asn Val Ile Thr Asn Gly Asp Phe Thr Gln Gly355 360 365
tta cag gga tgg cac gca aca gga aat gcc gcg gta caa caa atg gat 1152Leu Gln Gly Trp His Ala Thr Gly Asn Ala Ala Val Gln Gln Met Asp370 375 380gga gct tca gta tta gtt cta tca aat tgg agc gcg ggg gta tct caa 1200Gly Ala Ser Val Leu Val Leu Ser Asn Trp Ser Ala Gly Val Ser Gln385 390 395 400aac ttg cat gct caa gat cat cat gga tat gtg tta cgt gtg att gcc 1248Asn Leu His Ala Gln Asp His His Gly Tyr Val Leu Arg Val Ile Ala405 410 415aaa aaa gaa gga cct gga aaa ggg tat gta acg atg atg gat tgt aat 1296Lys Lys Glu Gly Pro Gly Lys Gly Tyr Val Thr Met Met Asp Cys Asn420 425 430gga aag cag gaa aca ctt aag ttc act tct tgc gaa gaa gga tat atg 1344Gly Lys Gln Glu Thr Leu Lys Phe Thr Ser Cys Glu Glu Gly Tyr Met435 440 445aca aaa aca gta gag gta ttc cca gaa agt gat cgt gta cgg att gaa 1392Thr Lys Thr Val Glu Val Phe Pro Glu Ser Asp Arg Val Arg Ile Glu450 455 460ata gga gaa acc gaa ggt aca ttt tat ata gat agc atc gag ttg ctt 1440Ile Gly Glu Thr Glu Gly Thr Phe Tyr Ile Asp Ser Ile Glu Leu Leu465 470 475 480tgt atg caa gga tat gat aac aat aat aac ctg cac acg ggt aat atg 1488Cys Met Gln Gly Tyr Asp Asn Asn Asn Asn Leu His Thr Gly Asn Met485 490 495tat gag caa agt tat aat gga aat tat aat caa aat act agc gat gtg 1536Tyr Glu Gln Ser Tyr Asn Gly Asn Tyr Asn Gln Asn Thr Ser Asp Val500 505 510tat tac caa ggg tat aca aac aac tat aac caa gac tct agt aat atg 1584Tyr Tyr Gln Gly Tyr Thr Asn Asn Tyr Asn Gln Asp Ser Ser Asn Met515 520 525tat aat caa aat tat act aac aat gat gac ctg cat tcc ggt tgc aca 1632Tyr Asn Gln Asn Tyr Thr Asn Asn Asp Asp Leu His Ser Gly Cys Thr530 535 540tgt aac caa ggg cat aac tct ggc tgt aca tgt aat caa gga tat aac 1680Cys Asn Gln Gly His Asn Ser Gly Cys Thr Cys Asn Gln Gly Tyr Asn545 550 555 560cgt taa 1686Arg *<210>18<211>561<212>PRT<213>Bacillus thuringiensis<400>18Met Ser Pro Met Phe Thr Ser Ser Thr Lys Asn Thr Leu Lys Ile Glu1 5 10 15Thr Thr Asp Tyr Glu Ile Asp Gln Ala Ala Ile Ser Ile Glu Cys Met20 25 30Ser Asp Glu Gln Asn Pro Gln Glu Lys Ile Met Leu Trp Asp Glu Ile35 40 45Lys Leu Ala Lys Gln Leu Ser Gln Ser Arg Asn Leu Leu Gln Asn Gly50 55 60Asp Phe Ser Gly Asn Asp Trp Thr Phe Gly Asn Asp Ile Ile Ile Gly
65 70 75 80Ser Asn Asn Pro Ile Phe Lys Gly Lys Phe Leu Gln Met Arg Gly Ala85 90 95Arg Asp Ile Tyr Gly Thr Leu Phe Pro Thr Tyr Ile Cys Gln Lys Ile100 105 110Asp Glu Ser Lys Leu Lys Pro Tyr Thr Arg Tyr Arg Val Arg Gly Phe115 120 125Val Gly Ser Ser Lys Asp Leu Lys Leu Met Val Thr Arg Tyr Gly Lys130 135 140Glu Ile Asp Ala Ile Met Asn Val Pro Asn Asp Leu Ala Tyr Met Gln145 150 155 160Pro Asn Pro Ser Cys Gly Asp Tyr Arg Cys Glu Ser Ser Ser Gln Tyr165 170 175Val Ser Gln Gly Tyr Pro Thr Pro Thr Asp Gly Tyr Ala Pro Asp Met180 185 190Tyr Ala Cys Pro Gln Asn Ile Asp Arg Lys His Val Lys Cys His Asp195 200 205Arg His Pro Phe Asp Phe His Ile Asp Thr Gly Glu Val Asp Thr Asn210 215 220Thr Asn Val Gly Ile Asp Val Leu Leu Lys Ile Ser Asn Pro Asp Gly225 230 235 240Tyr Ala Thr Val Gly Asn Leu Glu Val Ile Glu Glu Gly Pro Leu Thr245 250 255Gly Glu Ala Leu Ala His Val Lys Gln Lys Glu Lys Lys Trp Lys Gln260 265 270His Met Glu Lys Lys Arg Trp Glu Thr Gln Gln Ala Tyr Asp Pro Ala275 280 285Lys Gln Ala Val Asp Ala Leu Phe Thr Asn Glu Gln Glu Leu His Tyr290 295 300His Ile Thr Leu Asp His Ile Gln Asn Ala Asp Arg Leu Val Gln Ser305 310 315 320Ile Pro Tyr Val Tyr His Asn Trp Leu Pro Asn Ala Pro Gly Met Asn325 330 335Tyr Asp Val Tyr Gln Glu Leu Asn Ala Arg Ile Met Gln Gly Tyr Asn340 345 350Leu Tyr Asp Ala Arg Asn Val Ile Thr Asn Gly Asp Phe Thr Gln Gly355 360 365Leu Gln Gly Trp His Ala Thr Gly Asn Ala Ala Val Gln Gln Met Asp370 375 380Gly Ala Ser Val Leu Val Leu Ser Asn Trp Ser Ala Gly Val Ser Gln385 390 395 400Asn Leu His Ala Gln Asp His His Gly Tyr Val Leu Arg Val Ile Ala405 410 415Lys Lys Glu Gly Pro Gly Lys Gly Tyr Val Thr Met Met Asp Cys Asn420 425 430Gly Lys Gln Glu Thr Leu Lys Phe Thr Ser Cys Glu Glu Gly Tyr Met435 440 445Thr Lys Thr Val Glu Val Phe Pro Glu Ser Asp Arg Val Arg Ile Glu450 455 460Ile Gly Glu Thr Glu Gly Thr Phe Tyr Ile Asp Ser Ile Glu Leu Leu465 470 475 480Cys Met Gln Gly Tyr Asp Asn Asn Asn Asn Leu His Thr Gly Asn Met485 490 495Tyr Glu Gln Ser Tyr Asn Gly Asn Tyr Asn Gln Asn Thr Ser Asp Val500 505 510Tyr Tyr Gln Gly Tyr Thr Asn Asn Tyr Asn Gln Asp Ser Ser Asn Met515 520 525Tyr Asn Gln Asn Tyr Thr Asn Asn Asp Asp Leu His Ser Gly Cys Thr530 535 540Cys Asn Gln Gly His Asn Ser Gly Cys Thr Cys Asn Gln Gly Tyr Asn545 550 555 560Arg<210>19<211>2049<212>DNA
<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2049)<400>19atg aat tca tat aaa aat aaa aat gaa tat gaa atg ttg gat gct tta 48Met Asn Ser Tyr Lys Asn Lys Asn Glu Tyr Glu Met Leu Asp Ala Leu1 5 10 15cga atc aac tct aat atg tct aat tgt tat cca agg tat cca cta gca 96Arg Ile Asn Ser Asn Met Ser Asn Cys Tyr Pro Arg Tyr Pro Leu Ala20 25 30aaa gat cca caa atg act atg cga aac acg aac tat aaa gaa tgg cta 144Lys Asp Pro Gln Met Thr Met Arg Asn Thr Asn Tyr Lys Glu Trp Leu35 40 45aat atg tgt gat tca aat aca caa ttt att ggt gat ata agc acg tat 192Asn Met Cys Asp Ser Asn Thr Gln Phe Ile Gly Asp Ile Ser Thr Tyr50 55 60tct agc cct gaa gct gct tta agt gta cga gat gct gtt tta acg ggt 240Ser Ser Pro Glu Ala Ala Leu Ser Val Arg Asp Ala Val Leu Thr Gly65 70 75 80att aac agt gta ggg act ata ctt tcg aat tta ggg gtc cct ttg gca 288Ile Asn Ser Val Gly Thr Ile Leu Ser Asn Leu Gly Val Pro Leu Ala85 90 95agt caa tca ttt gga ata att agt agg cta ata ggt att tta tgg gca 336Ser Gln Ser Phe Gly Ile Ile Ser Arg Leu Ile Gly Ile Leu Trp Ala100 105 110ggg cct gat cca ttt gaa gca ctt atg gtt ctt gtt gaa gag ctt att 384Gly Pro Asp Pro Phe Glu Ala Leu Met Val Leu Val Glu Glu Leu Ile115 120 125aag aaa agt ata gat cag cgt gta aga gaa aat gct ctt aga gag cta 432Lys Lys Ser Ile Asp Gln Arg Val Arg Glu Asn Ala Leu Arg Glu Leu130 135 140gaa ggt tta cag gga att atg aga cta tat caa act aga ctg caa gca 480Glu Gly Leu Gln Gly Ile Met Arg Leu Tyr Gln Thr Arg Leu Gln Ala145 150 155 160tgg cta gtt aac aag aat gat gac aat cgg agg gca cta gta acg cag 528Trp Leu Val Asn Lys Asn Asp Asp Asn Arg Arg Ala Leu Val Thr Gln165 170 175tat gca att gtt gat aac ttt ttc gaa aag aat atg cca aaa ttc aag 576Tyr Ala Ile Val Asp Asn Phe Phe Glu Lys Asn Met Pro Lys Phe Lys180 185 190gaa aga aac ttt gaa att tta ttg tta cca gta tat gca caa gcc gcg 624Glu Arg Asn Phe Glu Ile Leu Leu Leu Pro Val Tyr Ala Gln Ala Ala195 200 205aat ttg cat tta att tta tta aga gat gct gat tat ttt gga gca cag 672Asn Leu His Leu Ile Leu Leu Arg Asp Ala Asp Tyr Phe Gly Ala Gln210 215 220tgg caa tta ggt gat gat gaa att cgt gat aat tat atc aga cta caa 720Trp Gln Leu Gly Asp Asp Glu Ile Arg Asp Asn Tyr Ile Arg Leu Gln225 230 235 240gga ctg att aga gaa tat aaa gat cat tgt ata aca ttc tat aac cag 768
Gly Leu Ile Arg Glu Tyr Lys Asp His Cys Ile Thr Phe Tyr Asn Gln245 250 255ggt tta aat caa ttt aat cgc tca aat gct caa gat tgg gtg agc ttt 816Gly Leu Asn Gln Phe Asn Arg Ser Asn Ala Gln Asp Trp Val Ser Phe260 265 270aat agg ttt cgt aca gat atg aca tta aca gta tta gat ctc gca ata 864Asn Arg Phe Arg Thr Asp Met Thr Leu Thr Val Leu Asp Leu Ala Ile275 280 285tta ttt cca aac tat gat cca cgt agg tat cca tta gca gta aaa acg 912Leu Phe Pro Asn Tyr Asp Pro Arg Arg Tyr Pro Leu Ala Val Lys Thr290 295 300gaa ttg act agg gaa gtt tat aca gat cca gta ggg ttt act ggg gta 960Glu Leu Thr Arg Glu Val Tyr Thr Asp Pro Val Gly Phe Thr Gly Val305 310 315 320tta gaa agt gga ggt agg act tac cct tgg tat aat cct aat aat aca 1008Leu Glu Ser Gly Gly Arg Thr Tyr Pro Trp Tyr Asn Pro Asn Asn Thr325 330 335acc ttt act gct atg gaa aat aac gca aga cga cgt cct tct tat acc 1056Thr Phe Thr Ala Met Glu Asn Asn Ala Arg Arg Arg Pro Ser Tyr Thr340 345 350act tgg ctt aat cgt att ttt gta tat aca agg act cta ggt aat atg 1104Thr Trp Leu Asn Arg Ile Phe Val Tyr Thr Arg Thr Leu Gly Asn Met355 360 365tct gat gtg aga aat att tgg gga ggg cat aca tta gtt gaa aat gga 1152Ser Asp Val Arg Asn Ile Trp Gly Gly His Thr Leu Val Glu Asn Gly370 375 380aat gat ggt tct gaa ata acc cat aac ttt ggt aaa act gat tct att 1200Asn Asp Gly Ser Glu Ile Thr His Asn Phe Gly Lys Thr Asp Ser Ile385 390 395 400act cct att caa tat ttt aat ttc gcg aac ctt tct gtt ttc agt att 1248Thr Pro Ile Gln Tyr Phe Asn Phe Ala Asn Leu Ser Val Phe Ser Ile405 410 415gag tca ctt gct cgt ata tat tta gga gga aca gag gct aat aat tat 1296Glu Ser Leu Ala Arg Ile Tyr Leu Gly Gly Thr Glu Ala Asn Asn Tyr420 425 430att act agt cag tat gga gtc tcg aga gtt att ttt aat aca tca aat 1344Ile Thr Ser Gln Tyr Gly Val Ser Arg Val Ile Phe Asn Thr Ser Asn435 440 445ata aat aat gta cct gga tct tta aga tac gaa gtg cct gct aat ctt 1392Ile Asn Asn Val Pro Gly Ser Leu Arg Tyr Glu Val Pro Ala Asn Leu450 455 460cca tcc caa act ata tta tca gaa tta cca gga aag gat aag cca aga 1440Pro Ser Gln Thr Ile Leu Ser Glu Leu Pro Gly Lys Asp Lys Pro Arg465 470 475 480cca aac gca gga gat ttc agc cat aga tta tct tat ata tca aat ttt 1488Pro Asn Ala Gly Asp Phe Ser His Arg Leu Ser Tyr Ile Ser Asn Phe485 490 495gat gca cgg cga agt agt tca ggc ggt att gtt agt ctt tta acg ttt 1536Asp Ala Arg Arg Ser Ser Ser Gly Gly Ile Val Ser Leu Leu Thr Phe500 505 510ggt tgg gca cat acc agt atg gat cgt aat aat cgt ctt gaa cca gat 1584
Gly Trp Ala His Thr Ser Met Asp Arg Asn Asn Arg Leu Glu Pro Asp515 520 525aaa att act caa ata gat gca gtt aaa ggt tgg ggg ggg aat atc ggg 1632Lys Ile Thr Gln Ile Asp Ala Val Lys Gly Trp Gly Gly Asn Ile Gly530 535 540ttt gtc atc cca gga cct act ggg ggg aat ttg gta aaa gtc agt gat 1680Phe Val Ile Pro Gly Pro Thr Gly Gly Asn Leu Val Lys Val Ser Asp545 550 555 560agt tgg cat tca ctt aaa gtt caa gca cca caa aga caa aca agt tat 1728Ser Trp His Ser Leu Lys Val Gln Ala Pro Gln Arg Gln Thr Ser Tyr565 570 575cgt att cgt ttg cgt tat gct tgt tta gtt acc cat ggg gat gct att 1776Arg Ile Arg Leu Arg Tyr Ala Cys Leu Val Thr His Gly Asp Ala Ile580 585 590ttt gta gaa cac agc ggc agt agt cat ata gtt tca ttt ttt gat tgc 1824Phe Val Glu His Ser Gly Ser Ser His Ile Val Ser Phe Phe Asp Cys595 600 605tca aat tca tca ggt cgt cca tca aac act ctt cta gag agt gat ttt 1872Ser Asn Ser Ser Gly Arg Pro Ser Asn Thr Leu Leu Glu Ser Asp Phe610 615 620cgc tat att gat gtt cca ggt att ttt aca cca tca ata aat ccc tta 1920Arg Tyr Ile Asp Val Pro Gly Ile Phe Thr Pro Ser Ile Asn Pro Leu625 630 635 640ata aga tat aga aca caa agc ttt ggt acc cac gcg ata gac aaa ttt 1968Ile Arg Tyr Arg Thr Gln Ser Phe Gly Thr His Ala Ile Asp Lys Phe645 650 655gaa ttt att cca ctt aac act ttt ccg aat caa tca tta gaa aaa aga 2016Glu Phe Ile Pro Leu Asn Thr Phe Pro Asn Gln Ser Leu Glu Lys Arg660 665 670gaa cag gaa gta aat gat cta ttt atc aat taa 2049Glu Gln Glu Val Asn Asp Leu Phe Ile Asn *675 680<210>20<211>682<212>PRT<213>Bacillus thuringiensis<400>20Met Asn Ser Tyr Lys Asn Lys Asn Glu Tyr Glu Met Leu Asp Ala Leu1 5 10 15Arg Ile Asn Ser Asn Met Ser Asn Cys Tyr Pro Arg Tyr Pro Leu Ala20 25 30Lys Asp Pro Gln Met Thr Met Arg Asn Thr Asn Tyr Lys Glu Trp Leu35 40 45Asn Met Cys Asp Ser Asn Thr Gln Phe Ile Gly Asp Ile Ser Thr Tyr50 55 60Ser Ser Pro Glu Ala Ala Leu Ser Val Arg Asp Ala Val Leu Thr Gly65 70 75 80Ile Asn Ser Val Gly Thr Ile Leu Ser Asn Leu Gly Val Pro Leu Ala85 90 95Ser Gln Ser Phe Gly Ile Ile Ser Arg Leu Ile Gly Ile Leu Trp Ala100 105 110Gly Pro Asp Pro Phe Glu Ala Leu Met Val Leu Val Glu Glu Leu Ile115 120 125Lys Lys Ser Ile Asp Gln Arg Val Arg Glu Asn Ala Leu Arg Glu Leu130 135 140
Glu Gly Leu Gln Gly Ile Met Arg Leu Tyr Gln Thr Arg Leu Gln Ala145 150 155 160Trp Leu Val Asn Lys Asn Asp Asp Asn Arg Arg Ala Leu Val Thr Gln165 170 175Tyr Ala Ile Val Asp Asn Phe Phe Glu Lys Asn Met Pro Lys Phe Lys180 185 190Glu Arg Asn Phe Glu Ile Leu Leu Leu Pro Val Tyr Ala Gln Ala Ala195 200 205Asn Leu His Leu Ile Leu Leu Arg Asp Ala Asp Tyr Phe Gly Ala Gln210 215 220Trp Gln Leu Gly Asp Asp Glu Ile Arg Asp Asn Tyr Ile Arg Leu Gln225 230 235 240Gly Leu Ile Arg Glu Tyr Lys Asp His Cys Ile Thr Phe Tyr Asn Gln245 250 255Gly Leu Asn Gln Phe Asn Arg Ser Asn Ala Gln Asp Trp Val Ser Phe260 265 270Asn Arg Phe Arg Thr Asp Met Thr Leu Thr Val Leu Asp Leu Ala Ile275 280 285Leu Phe Pro Asn Tyr Asp Pro Arg Arg Tyr Pro Leu Ala Val Lys Thr290 295 300Glu Leu Thr Arg Glu Val Tyr Thr Asp Pro Val Gly Phe Thr Gly Val305 310 315 320Leu Glu Ser Gly Gly Arg Thr Tyr Pro Trp Tyr Asn Pro Asn Asn Thr325 330 335Thr Phe Thr Ala Met Glu Asn Asn Ala Arg Arg Arg Pro Ser Tyr Thr340 345 350Thr Trp Leu Asn Arg Ile Phe Val Tyr Thr Arg Thr Leu Gly Asn Met355 360 365Ser Asp Val Arg Asn Ile Trp Gly Gly His Thr Leu Val Glu Asn Gly370 375 380Asn Asp Gly Ser Glu Ile Thr His Asn Phe Gly Lys Thr Asp Ser Ile385 390 395 400Thr Pro Ile Gln Tyr Phe Asn Phe Ala Asn Leu Ser Val Phe Ser Ile405 410 415Glu Ser Leu Ala Arg Ile Tyr Leu Gly Gly Thr Glu Ala Asn Asn Tyr420 425 430Ile Thr Ser Gln Tyr Gly Val Ser Arg Val Ile Phe Asn Thr Ser Asn435 440 445Ile Asn Asn Val Pro Gly Ser Leu Arg Tyr Glu Val Pro Ala Asn Leu450 455 460Pro Ser Gln Thr Ile Leu Ser Glu Leu Pro Gly Lys Asp Lys Pro Arg465 470 475 480Pro Asn Ala Gly Asp Phe Ser His Arg Leu Ser Tyr Ile Ser Asn Phe485 490 495Asp Ala Arg Arg Ser Ser Ser Gly Gly Ile Val Ser Leu Leu Thr Phe500 505 510Gly Trp Ala His Thr Ser Met Asp Arg Asn Asn Arg Leu Glu Pro Asp515 520 525Lys Ile Thr Gln Ile Asp Ala Val Lys Gly Trp Gly Gly Asn Ile Gly530 535 540Phe Val Ile Pro Gly Pro Thr Gly Gly Asn Leu Val Lys Val Ser Asp545 550 555 560Ser Trp His Ser Leu Lys Val Gln Ala Pro Gln Arg Gln Thr Ser Tyr565 570 575Arg Ile Arg Leu Arg Tyr Ala Cys Leu Val Thr His Gly Asp Ala Ile580 585 590Phe Val Glu His Ser Gly Ser Ser His Ile Val Ser Phe Phe Asp Cys595 600 605Ser Asn Ser Ser Gly Arg Pro Ser Asn Thr Leu Leu Glu Ser Asp Phe610 615 620Arg Tyr Ile Asp Val Pro Gly Ile Phe Thr Pro Ser Ile Asn Pro Leu625 630 635 640Ile Arg Tyr Arg Thr Gln Ser Phe Gly Thr His Ala Ile Asp Lys Phe645 650 655Glu Phe Ile Pro Leu Asn Thr Phe Pro Asn Gln Ser Leu Glu Lys Arg660 665 670Glu Gln Glu Val Asn Asp Leu Phe Ile Asn675 680
<210>21<211>2016<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2016)<A00>21atg ttg gat gct tta cga atc aac tct aat atg tct aat tgt tat cca 48Met Leu Asp Ala Leu Arg Ile Asn Ser Asn Met Ser Asn Cys Tyr Pro1 5 10 15agg tat cca cta gca aaa gat cca caa atg act atg cga aac acg aac 96Arg Tyr Pro Leu Ala Lys Asp Pro Gln Met Thr Met Arg Asn Thr Asn20 25 30tat aaa gaa tgg cta aat atg tgt gat tca aat aca caa ttt att ggt 144Tyr Lys Glu Trp Leu Asn Met Cys Asp Ser Asn Thr Gln Phe Ile Gly35 40 45gat ata agc acg tat tct agc cct gaa gct gct tta agt gta cga gat 192Asp Ile Ser Thr Tyr Ser Ser Pro Glu Ala Ala Leu Ser Val Arg Asp50 55 60gct gtt tta acg ggt att aac agt gta ggg act ata ctt tcg aat tta 240Ala Val Leu Thr Gly Ile Asn Ser Val Gly Thr Ile Leu Ser Asn Leu65 70 75 80ggg gtc cct ttg gca agt caa tca ttt gga ata att agt agg cta ata 288Gly Val Pro Leu Ala Ser Gln Ser Phe Gly Ile Ile Ser Arg Leu Ile85 90 95ggt att tta tgg gca ggg cct gat cca ttt gaa gca ctt atg gtt ctt 336Gly Ile Leu Trp Ala Gly Pro Asp Pro Phe Glu Ala Leu Met Val Leu100 105 110gtt gaa gag ctt att aag aaa agt ata gat cag cgt gta aga gaa aat 384Val Glu Glu Leu Ile Lys Lys Ser Ile Asp Gln Arg Val Arg Glu Asn115 120 125gct ctt aga gag cta gaa ggt tta cag gga att atg aga cta tat caa 432Ala Leu Arg Glu Leu Glu Gly Leu Gln Gly Ile Met Arg Leu Tyr Gln130 135 140act aga ctg caa gca tgg cta gtt aac aag aat gat gac aat cgg agg 480Thr Arg Leu Gln Ala Trp Leu Val Asn Lys Asn Asp Asp Asn Arg Arg145 150 155 160gca cta gta acg cag tat gca att gtt gat aac ttt ttc gaa aag aat 528Ala Leu Val Thr Gln Tyr Ala Ile Val Asp Asn Phe Phe Glu Lys Asn165 170 175atg cca aaa ttc aag gaa aga aac ttt gaa att tta ttg tta cca gta 576Met Pro Lys Phe Lys Glu Arg Asn Phe Glu Ile Leu Leu Leu Pro Val180 185 190tat gca caa gcc gcg aat ttg cat tta att tta tta aga gat gct gat 624Tyr Ala Gln Ala Ala Asn Leu His Leu Ile Leu Leu Arg Asp Ala Asp195 200 205tat ttt gga gca cag tgg caa tta ggt gat gat gaa att cgt gat aat 672Tyr Phe Gly Ala Gln Trp Gln Leu Gly Asp Asp Glu Ile Arg Asp Asn210 215 220
tat atc aga cta caa gga ctg att aga gaa tat aaa gat cat tgt ata 720Tyr Ile Arg Leu Gln Gly Leu Ile Arg Glu Tyr Lys Asp His Cys Ile225 230 235 240aca ttc tat aac cag ggt tta aat caa ttt aat cgc tca aat gct caa 768Thr Phe Tyr Asn Gln Gly Leu Asn Gln Phe Asn Arg Ser Asn Ala Gln245 250 255gat tgg gtg agc ttt aat agg ttt cgt aca gat atg aca tta aca gta 816Asp Trp Val Ser Phe Asn Arg Phe Arg Thr Asp Met Thr Leu Thr Val260 265 270tta gat ctc gca ata tta ttt cca aac tat gat cca cgt agg tat cca 864Leu Asp Leu Ala Ile Leu Phe Pro Asn Tyr Asp Pro Arg Arg Tyr Pro275 280 285tta gca gta aaa acg gaa ttg act agg gaa gtt tat aca gat cca gta 912Leu Ala Val Lys Thr Glu Leu Thr Arg Glu Val Tyr Thr Asp Pro Val290 295 300ggg ttt act ggg gta tta gaa agt gga ggt agg act tac cct tgg tat 960Gly Phe Thr Gly Val Leu Glu Ser Gly Gly Arg Thr Tyr Pro Trp Tyr305 310 315 320aat cct aat aat aca acc ttt act gct atg gaa aat aac gca aga cga 1008Asn Pro Asn Asn Thr Thr Phe Thr Ala Met Glu Asn Asn Ala Arg Arg325 330 335cgt cct tct tat acc act tgg ctt aat cgt att ttt gta tat aca agg 1056Arg Pro Ser Tyr Thr Thr Trp Leu Asn Arg Ile Phe Val Tyr Thr Arg340 345 350act cta ggt aat atg tct gat gtg aga aat att tgg gga ggg cat aca 1104Thr Leu Gly Asn Met Ser Asp Val Arg Asn Ile Trp Gly Gly His Thr355 360 365tta gtt gaa aat gga aat gat ggt tct gaa ata acc cat aac ttt ggt 1152Leu Val Glu Asn Gly Asn Asp Gly Ser Glu Ile Thr His Asn Phe Gly370 375 380aaa act gat tct att act cct att caa tat ttt aat ttc gcg aac ctt 1200Lys Thr Asp Ser Ile Thr Pro Ile Gln Tyr Phe Asn Phe Ala Asn Leu385 390 395 400tct gtt ttc agt att gag tca ctt gct cgt ata tat tta gga gga aca 1248Ser Val Phe Ser Ile Glu Ser Leu Ala Arg Ile Tyr Leu Gly Gly Thr405 410 415gag gct aat aat tat att act agt cag tat gga gtc tcg aga gtt att 1296Glu Ala Asn Asn Tyr Ile Thr Ser Gln Tyr Gly Val Ser Arg Val Ile420 425 430ttt aat aca tca aat ata aat aat gta cct gga tct tta aga tac gaa 1344Phe Asn Thr Ser Asn Ile Asn Asn Val Pro Gly Ser Leu Arg Tyr Glu435 440 445gtg cct gct aat ctt cca tcc caa act ata tta tca gaa tta cca gga 1392Val Pro Ala Asn Leu Pro Ser Gln Thr Ile Leu Ser Glu Leu Pro Gly450 455 460aag gat aag cca aga cca aac gca gga gat ttc agc cat aga tta tct 1440Lys Asp Lys Pro Arg Pro Asn Ala Gly Asp Phe Ser His Arg Leu Ser465 470 475 480tat ata tca aat ttt gat gca cgg cga agt agt tca ggc ggt att gtt 1488Tyr Ile Ser Asn Phe Asp Ala Arg Arg Ser Ser Ser Gly Gly Ile Val485 490 495
agt ctt tta acg ttt ggt tgg gca cat acc agt atg gat cgt aat aat 1536Ser Leu Leu Thr Phe Gly Trp Ala His Thr Ser Met Asp Arg Asn Asn500 505 510cgt ctt gaa cca gat aaa att act caa ata gat gca gtt aaa ggt tgg 1584Arg Leu Glu Pro Asp Lys Ile Thr Gln Ile Asp Ala Val Lys Gly Trp515 520 525ggg ggg aat atc ggg ttt gtc atc cca gga cct act ggg ggg aat ttg 1632Gly Gly Asn Ile Gly Phe Val Ile Pro Gly Pro Thr Gly Gly Asn Leu530 535 540gta aaa gtc agt gat agt tgg cat tca ctt aaa gtt caa gca cca caa 1680Val Lys Val Ser Asp Ser Trp His Ser Leu Lys Val Gln Ala Pro Gln545 550 555 560aga caa aca agt tat cgt att cgt ttg cgt tat gct tgt tta gtt acc 1728Arg Gln Thr Ser Tyr Arg Ile Arg Leu Arg Tyr Ala Cys Leu Val Thr565 570 575cat ggg gat gct att ttt gta gaa cac agc ggc agt agt cat ata gtt 1776His Gly Asp Ala Ile Phe Val Glu His Ser Gly Ser Ser His Ile Val580 585 590tca ttt ttt gat tgc tca aat tca tca ggt cgt cca tca aac act ctt 1824Ser Phe Phe Asp Cys Ser Asn Ser Ser Gly Arg Pro Ser Asn Thr Leu595 600 605cta gag agt gat ttt cgc tat att gat gtt cca ggt att ttt aca cca 1872Leu Glu Ser Asp Phe Arg Tyr Ile Asp Val Pro Gly Ile Phe Thr Pro610 615 620tca ata aat ccc tta ata aga tat aga aca caa agc ttt ggt acc cac 1920Ser Ile Asn Pro Leu Ile Arg Tyr Arg Thr Gln Ser Phe Gly Thr His625 630 635 640gcg ata gac aaa ttt gaa ttt att cca ctt aac act ttt ccg aat caa 1968Ala Ile Asp Lys Phe Glu Phe Ile Pro Leu Asn Thr Phe Pro Asn Gln645 650 655tca tta gaa aaa aga gaa cag gaa gta aat gat cta ttt atc aat taa 2016Ser Leu Glu Lys Arg Glu Gln Glu Val Asn Asp Leu Phe Ile Asn *660 665 670<210>22<211>671<212>PRT<213>Bacillus thuringiensis<400>22Met Leu Asp Ala Leu Arg Ile Asn Ser Asn Met Ser Asn Cys Tyr Pro1 5 10 15Arg Tyr Pro Leu Ala Lys Asp Pro Gln Met Thr Met Arg Asn Thr Asn20 25 30Tyr Lys Glu Trp Leu Asn Met Cys Asp Ser Asn Thr Gln Phe Ile Gly35 40 45Asp Ile Ser Thr Tyr Ser Ser Pro Glu Ala Ala Leu Ser Val Arg Asp50 55 60Ala Val Leu Thr Gly Ile Asn Ser Val Gly Thr Ile Leu Ser Asn Leu65 70 75 80Gly Val Pro Leu Ala Ser Gln Ser Phe Gly Ile Ile Ser Arg Leu Ile85 90 95Gly Ile Leu Trp Ala Gly Pro Asp Pro Phe Glu Ala Leu Met Val Leu100 105 110Val Glu Glu Leu Ile Lys Lys Ser Ile Asp Gln Arg Val Arg Glu Asn115 120 125
Ala Leu Arg Glu Leu Glu Gly Leu Gln Gly Ile Met Arg Leu Tyr Gln130 135 140Thr Arg Leu Gln Ala Trp Leu Val Asn Lys Asn Asp Asp Asn Arg Arg145 150 155 160Ala Leu Val Thr Gln Tyr Ala Ile Val Asp Asn Phe Phe Glu Lys Asn165 170 175Met Pro Lys Phe Lys Glu Arg Asn Phe Glu Ile Leu Leu Leu Pro Val180 185 190Tyr Ala Gln Ala Ala Asn Leu His Leu Ile Leu Leu Arg Asp Ala Asp195 200 205Tyr Phe Gly Ala Gln Trp Gln Leu Gly Asp Asp Glu Ile Arg Asp Asn210 215 220Tyr Ile Arg Leu Gln Gly Leu Ile Arg Glu Tyr Lys Asp His Cys Ile225 230 235 240Thr Phe Tyr Asn Gln Gly Leu Asn Gln Phe Asn Arg Ser Asn Ala Gln245 250 255Asp Trp Val Ser Phe Asn Arg Phe Arg Thr Asp Met Thr Leu Thr Val260 265 270Leu Asp Leu Ala Ile Leu Phe Pro Asn Tyr Asp Pro Arg Arg Tyr Pro275 280 285Leu Ala Val Lys Thr Glu Leu Thr Arg Glu Val Tyr Thr Asp Pro Val290 295 300Gly Phe Thr Gly Val Leu Glu Ser Gly Gly Arg Thr Tyr Pro Trp Tyr305 310 315 320Asn Pro Asn Asn Thr Thr Phe Thr Ala Met Glu Asn Asn Ala Arg Arg325 330 335Arg Pro Ser Tyr Thr Thr Trp Leu Asn Arg Ile Phe Val Tyr Thr Arg340 345 350Thr Leu Gly Asn Met Ser Asp Val Arg Asn Ile Trp Gly Gly His Thr355 360 365Leu Val Glu Asn Gly Asn Asp Gly Ser Glu Ile Thr His Asn Phe Gly370 375 380Lys Thr Asp Ser Ile Thr Pro Ile Gln Tyr Phe Asn Phe Ala Asn Leu385 390 395 400Ser Val Phe Ser Ile Glu Ser Leu Ala Arg Ile Tyr Leu Gly Gly Thr405 410 415Glu Ala Asn Asn Tyr Ile Thr Ser Gln Tyr Gly Val Ser Arg Val Ile420 425 430Phe Asn Thr Ser Asn Ile Asn Asn Val Pro Gly Ser Leu Arg Tyr Glu435 440 445Val Pro Ala Asn Leu Pro Ser Gln Thr Ile Leu Ser Glu Leu Pro Gly450 455 460Lys Asp Lys Pro Arg Pro Asn Ala Gly Asp Phe Ser His Arg Leu Ser465 470 475 480Tyr Ile Ser Asn Phe Asp Ala Arg Arg Ser Ser Ser Gly Gly Ile Val485 490 495Ser Leu Leu Thr Phe Gly Trp Ala His Thr Ser Met Asp Arg Asn Asn500 505 510Arg Leu Glu Pro Asp Lys Ile Thr Gln Ile Asp Ala Val Lys Gly Trp515 520 525Gly Gly Asn Ile Gly Phe Val Ile Pro Gly Pro Thr Gly Gly Asn Leu530 535 540Val Lys Val Ser Asp Ser Trp His Ser Leu Lys Val Gln Ala Pro Gln545 550 555 560Arg Gln Thr Ser Tyr Arg Ile Arg Leu Arg Tyr Ala Cys Leu Val Thr565 570 575His Gly Asp Ala Ile Phe Val Glu His Ser Gly Ser Ser His Ile Val580 585 590Ser Phe Phe Asp Cys Ser Asn Ser Ser Gly Arg Pro Ser Asn Thr Leu595 600 605Leu Glu Ser Asp Phe Arg Tyr Ile Asp Val Pro Gly Ile Phe Thr Pro610 615 620Ser Ile Asn Pro Leu Ile Arg Tyr Arg Thr Gln Ser Phe Gly Thr His625 630 635 640Ala Ile Asp Lys Phe Glu Phe Ile Pro Leu Asn Thr Phe Pro Asn Gln645 650 655Ser Leu Glu Lys Arg Glu Gln Glu Val Asn Asp Leu Phe Ile Asn660 665 670
<210>23<211>1986<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(1986)<400>23atg tct aat tgt tat cca agg tat cca cta gca aaa gat cca caa atg 48Met Ser Asn Cys Tyr Pro Arg Tyr Pro Leu Ala Lys Asp Pro Gln Met1 5 10 15act atg cga aac acg aac tat aaa gaa tgg cta aat atg tgt gat tca 96Thr Met Arg Asn Thr Asn Tyr Lys Glu Trp Leu Asn Met Cys Asp Ser20 25 30aat aca caa ttt att ggt gat ata agc acg tat tct agc cct gaa gct 144Asn Thr Gln Phe Ile Gly Asp Ile Ser Thr Tyr Ser Ser Pro Glu Ala35 40 45gct tta agt gta cga gat gct gtt tta acg ggt att aac agt gta ggg 192Ala Leu Ser Val Arg Asp Ala Val Leu Thr Gly Ile Asn Ser Val Gly50 55 60act ata ctt tcg aat tta ggg gtc cct ttg gca agt caa tca ttt gga 240Thr Ile Leu Ser Asn Leu Gly Val Pro Leu Ala Ser Gln Ser Phe Gly65 70 75 80ata att agt agg cta ata ggt att tta tgg gca ggg cct gat cca ttt 288Ile Ile Ser Arg Leu Ile Gly Ile Leu Trp Ala Gly Pro Asp Pro Phe85 90 95gaa gca ctt atg gtt ctt gtt gaa gag ctt att aag aaa agt ata gat 336Glu Ala Leu Met Val Leu Val Glu Glu Leu Ile Lys Lys Ser Ile Asp100 105 110cag cgt gta aga gaa aat gct ctt aga gag cta gaa ggt tta cag gga 384Gln Arg Val Arg Glu Asn Ala Leu Arg Glu Leu Glu Gly Leu Gln Gly115 120 125att atg aga cta tat caa act aga ctg caa gca tgg cta gtt aac aag 432Ile Met Arg Leu Tyr Gln Thr Arg Leu Gln Ala Trp Leu Val Asn Lys130 135 140aat gat gac aat cgg agg gca cta gta acg cag tat gca att gtt gat 480Asn Asp Asp Asn Arg Arg Ala Leu Val Thr Gln Tyr Ala Ile Val Asp145 150 155 160aac ttt ttc gaa aag aat atg cca aaa ttc aag gaa aga aac ttt gaa 528Asn Phe Phe Glu Lys Asn Met Pro Lys Phe Lys Glu Arg Asn Phe Glu165 170 175att tta ttg tta cca gta tat gca caa gcc gcg aat ttg cat tta att 576Ile Leu Leu Leu Pro Val Tyr Ala Gln Ala Ala Asn Leu His Leu Ile180 185 190tta tta aga gat gct gat tat ttt gga gca cag tgg caa tta ggt gat 624Leu Leu Arg Asp Ala Asp Tyr Phe Gly Ala Gln Trp Gln Leu Gly Asp195 200 205gat gaa att cgt gat aat tat atc aga cta caa gga ctg att aga gaa 672Asp Glu Ile Arg Asp Asn Tyr Ile Arg Leu Gln Gly Leu Ile Arg Glu210 215 220
tat aaa gat cat tgt ata aca ttc tat aac cag ggt tta aat caa ttt 720Tyr Lys Asp His Cys Ile Thr Phe Tyr Asn Gln Gly Leu Asn Gln Phe225 230 235 240aat cgc tca aat gct caa gat tgg gtg agc ttt aat agg ttt cgt aca 768Asn Arg Ser Asn Ala Gln Asp Trp Val Ser Phe Asn Arg Phe Arg Thr245 250 255gat atg aca tta aca gta tta gat ctc gca ata tta ttt cca aac tat 816Asp Met Thr Leu Thr Val Leu Asp Leu Ala Ile Leu Phe Pro Asn Tyr260 265 270gat cca cgt agg tat cca tta gca gta aaa acg gaa ttg act agg gaa 864Asp Pro Arg Arg Tyr Pro Leu Ala Val Lys Thr Glu Leu Thr Arg Glu275 280 285gtt tat aca gat cca gta ggg ttt act ggg gta tta gaa agt gga ggt 912Val Tyr Thr Asp Pro Val Gly Phe Thr Gly Val Leu Glu Ser Gly Gly290 295 300agg act tac cct tgg tat aat cct aat aat aca acc ttt act gct atg 960Arg Thr Tyr Pro Trp Tyr Asn Pro Asn Asn Thr Thr Phe Thr Ala Met305 310 315 320gaa aat aac gca aga cga cgt cct tct tat acc act tgg ctt aat cgt 1008Glu Asn Asn Ala Arg Arg Arg Pro Ser Tyr Thr Thr Trp Leu Asn Arg325 330 335att ttt gta tat aca agg act cta ggt aat atg tct gat gtg aga aat 1056Ile Phe Val Tyr Thr Arg Thr Leu Gly Asn Met Ser Asp Val Arg Asn340 345 350att tgg gga ggg cat aca tta gtt gaa aat gga aat gat ggt tct gaa 1104Ile Trp Gly Gly His Thr Leu Val Glu Asn Gly Asn Asp Gly Ser Glu355 360 365ata acc cat aac ttt ggt aaa act gat tct att act cct att caa tat 1152Ile Thr His Asn Phe Gly Lys Thr Asp Ser Ile Thr Pro Ile Gln Tyr370 375 380ttt aat ttc gcg aac ctt tct gtt ttc agt att gag tca ctt gct cgt 1200Phe Asn Phe Ala Asn Leu Ser Val Phe Ser Ile Glu Ser Leu Ala Arg385 390 395 400ata tat tta gga gga aca gag gct aat aat tat att act agt cag tat 1248Ile Tyr Leu Gly Gly Thr Glu Ala Asn Asn Tyr Ile Thr Ser Gln Tyr405 410 415gga gtc tcg aga gtt att ttt aat aca tca aat ata aat aat gta cct 1296Gly Val Ser Arg Val Ile Phe Asn Thr Ser Asn Ile Asn Asn Val Pro420 425 430gga tct tta aga tac gaa gtg cct gct aat ctt cca tcc caa act ata 1344Gly Ser Leu Arg Tyr Glu Val Pro Ala Asn Leu Pro Ser Gln Thr Ile435 440 445tta tca gaa tta cca gga aag gat aag cca aga cca aac gca gga gat 1392Leu Ser Glu Leu Pro Gly Lys Asp Lys Pro Arg Pro Asn Ala Gly Asp450 455 460ttc agc cat aga tta tct tat ata tca aat ttt gat gca cgg cga agt 1440Phe Ser His Arg Leu Ser Tyr Ile Ser Asn Phe Asp Ala Arg Arg Ser465 470 475 480agt tca ggc ggt att gtt agt ctt tta acg ttt ggt tgg gca cat acc 1488Ser Ser Gly Gly Ile Val Ser Leu Leu Thr Phe Gly Trp Ala His Thr485 490 495
agt atg gat cgt aat aat cgt ctt gaa cca gat aaa att act caa ata 1536Ser Met Asp Arg Asn Asn Arg Leu Glu Pro Asp Lys Ile Thr Gln Ile500 505 510gat gca gtt aaa ggt tgg ggg ggg aat atc ggg ttt gtc atc cca gga 1584Asp Ala Val Lys Gly Trp Gly Gly Asn Ile Gly Phe Val Ile Pro Gly515 520 525cct act ggg ggg aat ttg gta aaa gtc agt gat agt tgg cat tca ctt 1632Pro Thr Gly Gly Asn Leu Val Lys Val Ser Asp Ser Trp His Ser Leu530 535 540aaa gtt caa gca cca caa aga caa aca agt tat cgt att cgt ttg cgt 1680Lys Val Gln Ala Pro Gln Arg Gln Thr Ser Tyr Arg Ile Arg Leu Arg545 550 555 560tat gct tgt tta gtt acc cat ggg gat gct att ttt gta gaa cac agc 1728Tyr Ala Cys Leu Val Thr His Gly Asp Ala Ile Phe Val Glu His Ser565 570 575ggc agt agt cat ata gtt tca ttt ttt gat tgc tca aat tca tca ggt 1776Gly Ser Ser His Ile Val Ser Phe Phe Asp Cys Ser Asn Ser Ser Gly580 585 590cgt cca tca aac act ctt cta gag agt gat ttt cgc tat att gat gtt 1824Arg Pro Ser Asn Thr Leu Leu Glu Ser Asp Phe Arg Tyr Ile Asp Val595 600 605cca ggt att ttt aca cca tca ata aat ccc tta ata aga tat aga aca 1872Pro Gly Ile Phe Thr Pro Ser Ile Asn Pro Leu Ile Arg Tyr Arg Thr610 615 620caa agc ttt ggt acc cac gcg ata gac aaa ttt gaa ttt att cca ctt 1920Gln Ser Phe Gly Thr His Ala Ile Asp Lys Phe Glu Phe Ile Pro Leu625 630 635 640aac act ttt ccg aat caa tca tta gaa aaa aga gaa cag gaa gta aat 1968Asn Thr Phe Pro Asn Gln Ser Leu Glu Lys Arg Glu Gln Glu Val Asn645 650 655gat cta ttt atc aat taa 1986Asp Leu Phe Ile Asn *660<210>24<211>661<212>PRT<213>Bacillus thuringiensis<400>24Met Ser Asn Cys Tyr Pro Arg Tyr Pro Leu Ala Lys Asp Pro Gln Met1 5 10 15Thr Met Arg Asn Thr Asn Tyr Lys Glu Trp Leu Asn Met Cys Asp Ser20 25 30Asn Thr Gln Phe Ile Gly Asp Ile Ser Thr Tyr Ser Ser Pro Glu Ala35 40 45Ala Leu Ser Val Arg Asp Ala Val Leu Thr Gly Ile Asn Ser Val Gly50 55 60Thr Ile Leu Ser Asn Leu Gly Val Pro Leu Ala Ser Gln Ser Phe Gly65 70 75 80Ile Ile Ser Arg Leu Ile Gly Ile Leu Trp Ala Gly Pro Asp Pro Phe85 90 95Glu Ala Leu Met Val Leu Val Glu Glu Leu Ile Lys Lys Ser Ile Asp100 105 110Gln Arg Val Arg Glu Asn Ala Leu Arg Glu Leu Glu Gly Leu Gln Gly115 120 125Ile Met Arg Leu Tyr Gln Thr Arg Leu Gln Ala Trp Leu Val Asn Lys
130 135 140Asn Asp Asp Asn Arg Arg Ala Leu Val Thr Gln Tyr Ala Ile Val Asp145 150 155 160Asn Phe Phe Glu Lys Asn Met Pro Lys Phe Lys Glu Arg Asn Phe Glu165 170 175Ile Leu Leu Leu Pro Val Tyr Ala Gln Ala Ala Asn Leu His Leu Ile180 185 190Leu Leu Arg Asp Ala Asp Tyr Phe Gly Ala Gln Trp Gln Leu Gly Asp195 200 205Asp Glu Ile Arg Asp Asn Tyr Ile Arg Leu Gln Gly Leu Ile Arg Glu210 215 220Tyr Lys Asp His Cys Ile Thr Phe Tyr Asn Gln Gly Leu Asn Gln Phe225 230 235 240Asn Arg Ser Asn Ala Gln Asp Trp Val Ser Phe Asn Arg Phe Arg Thr245 250 255Asp Met Thr Leu Thr Val Leu Asp Leu Ala Ile Leu Phe Pro Asn Tyr260 265 270Asp Pro Arg Arg Tyr Pro Leu Ala Val Lys Thr Glu Leu Thr Arg Glu275 280 285Val Tyr Thr Asp Pro Val Gly Phe Thr Gly Val Leu Glu Ser Gly Gly290 295 300Arg Thr Tyr Pro Trp Tyr Asn Pro Asn Asn Thr Thr Phe Thr Ala Met305 310 315 320Glu Asn Asn Ala Arg Arg Arg Pro Ser Tyr Thr Thr Trp Leu Asn Arg325 330 335Ile Phe Val Tyr Thr Arg Thr Leu Gly Asn Met Ser Asp Val Arg Asn340 345 350Ile Trp Gly Gly His Thr Leu Val Glu Asn Gly Asn Asp Gly Ser Glu355 360 365Ile Thr His Asn Phe Gly Lys Thr Asp Ser Ile Thr Pro Ile Gln Tyr370 375 380Phe Asn Phe Ala Asn Leu Ser Val Phe Ser Ile Glu Ser Leu Ala Arg385 390 395 400Ile Tyr Leu Gly Gly Thr Glu Ala Asn Asn Tyr Ile Thr Ser Gln Tyr405 410 415Gly Val Ser Arg Val Ile Phe Asn Thr Ser Asn Ile Asn Asn Val Pro420 425 430Gly Ser Leu Arg Tyr Glu Val Pro Ala Asn Leu Pro Ser Gln Thr Ile435 440 445Leu Ser Glu Leu Pro Gly Lys Asp Lys Pro Arg Pro Asn Ala Gly Asp450 455 460Phe Ser His Arg Leu Ser Tyr Ile Ser Asn Phe Asp Ala Arg Arg Ser465 470 475 480Ser Ser Gly Gly Ile Val Ser Leu Leu Thr Phe Gly Trp Ala His Thr485 490 495Ser Met Asp Arg Asn Asn Arg Leu Glu Pro Asp Lys Ile Thr Gln Ile500 505 510Asp Ala Val Lys Gly Trp Gly Gly Asn Ile Gly Phe Val Ile Pro Gly515 520 525Pro Thr Gly Gly Asn Leu Val Lys Val Ser Asp Ser Trp His Ser Leu530 535 540Lys Val Gln Ala Pro Gln Arg Gln Thr Ser Tyr Arg Ile Arg Leu Arg545 550 555 560Tyr Ala Cys Leu Val Thr His Gly Asp Ala Ile Phe Val Glu His Ser565 570 575Gly Ser Ser His Ile Val Ser Phe Phe Asp Cys Ser Asn Ser Ser Gly580 585 590Arg Pro Ser Asn Thr Leu Leu Glu Ser Asp Phe Arg Tyr Ile Asp Val595 600 605Pro Gly Ile Phe Thr Pro Ser Ile Asn Pro Leu Ile Arg Tyr Arg Thr610 615 620Gln Ser Phe Gly Thr His Ala Ile Asp Lys Phe Glu Phe Ile Pro Leu625 630 635 640Asn Thr Phe Pro Asn Gln Ser Leu Glu Lys Arg Glu Gln Glu Val Asn645 650 655Asp Leu Phe Ile Asn660
<210>25<211>2145<212>DNA<213>Bacillus thuringiensis<400>25tttcaccaaa tataatgttt ttttgtagaa acattgttac aggaatacat tggggtacta 60cgaatatata gaaagacacc tggcatatat ttattaggtg tcttaaaaat aaggactata 120taaggagtga aaaatatgaa ttcttatcaa aatacaaatg aatatgaaat tctggatggt 180tccccgaata acacaaatat gtcaaacaga tatccttttg caaaggatcc aaatatattt 240cctattaacc tggacgcttg tcagggaagg ccatggcaag atacgtggga atcagtctcg 300gatatagtaa ctattgggac ataccttata caattcttgc tagaacccgg tataggtgga 360attcctgtaa tattttcaat aataaacaaa ctcattccgt cttctggtca atctgtggca 420gcactttcta tatgtgattt agtatctata attcgtaaag aggtagacga gagcgtgtta 480agtgacgggg ttgcagattt tgagggtgaa atgactgctt atcaagatta ttatcttcat 540tatcttgagg attggcttac agataaatca aatcctaaaa aacttgctga cgtagttaaa 600cagttccaag cacgggaaga agatttcact aaacttttag cagggtcatt atcaagacag 660aaagctgaaa tattattatt gcctacgtat gtgcaagctg caaatgtgca tttattacta 720ttaagggacg cagttaaata taaaaaagaa tggggactag tgtgtccacc gttgtatcca 780gggtcaggga gaactgattg taacgagcgg ttaaaagcga aaataaaaga gtatactaat 840tattgtgtag ggtggtataa caagggttta gatcagataa gacaggcggg tacaagtgct 900gaagtttggt cgaaatttaa taaatttcgt agagaaatga cgttggcggt attggatatt 960attgctatat ttccaactta tgattttgaa aaatatccat tagcaacaag tgtagagtta 1020actagggaaa tttatacaga tccagtggga tattcagggg gaaattatgg ttgggaacgg 1080ttttttagct ttaattcggt agaagcaaat ggaacacggg gacctggttt agttacttgg 1140cttcaagcta tagatatata tagtcattct attaatcttc agcttggtta tcttagtggc 1200tgggggggaa ctcgtcatta tgaagacttc acaaagggta acggtgcttt tcaacgtatg 1260tctggaacta cgagtaataa tccacgtaat attatttttg gcaataccga tatatttaaa 1320attatttcat tagctagata tgcaatgcaa ccgtttgttg ggtattcaat cccacggcat 1380cttgtttcac gtgcagaatt ttttccgaca acactaaata ctttcctgta tgaggtaaac 1440agttctgggt actcacagac aattgaatct gtgttaccag gtattaataa ggatctacca 1500cctagtcgta caaattactc tcatagatta tcaaatgcgg catgtgttca aaatgaaacc 1560tccagagtta acgtatttgg ttggacacat acaagtatga aaaaagataa tcgaatttat 1620ccagataaaa ttacgcaaat tcctgcagta aaagcttttg ccctaccagc aggtacagga 1680tatgcaggag gttacgtcac agctgggcct ggttatacag gaggagatgt agtaacgtta 1740ccttatcaag caagtttaaa aatacgttta acttctgcac ccacgaataa aaattaccgt 1800gttagacttc gctacgcgag tggaggacct ggtccgttcc gagtagaaag atggtcgcca 1860agttctgttt caaatgctaa tttttctcgt ccagctacag gtggctatag ttcatttgat 1920tatgtggaca ccttagttac tacatttaat caatcaggtg ttgaaataat tatacaaaat 1980ctatctggtt accaccttat tgttgacaaa gtcgaattta tcccaattga catccaaatt 2040gaaaaatgta cgaaatgtca attcgaagga gacatatgta gatgtgaagg agtacaatcc 2100ttggaaacaa aaaaagagat tgtaaatagt ttatttatca attaa 2145<210>26<211>2019<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2019)<400>26gtg aaa aat atg aat tct tat caa aat aca aat gaa tat gaa att ctg 48Met Lys Asn Met Asn Ser Tyr Gln Asn Thr Asn Glu Tyr Glu Ile Leu1 5 10 15gat ggt tcc ccg aat aac aca aat atg tca aac aga tat cct ttt gca 96Asp Gly Ser Pro Asn Asn Thr Asn Met Ser Asn Arg Tyr Pro Phe Ala20 25 30aag gat cca aat ata ttt cct att aac ctg gac gct tgt cag gga agg 144Lys Asp Pro Asn Ile Phe Pro Ile Asn Leu Asp Ala Cys Gln Gly Arg35 40 45cca tgg caa gat acg tgg gaa tca gtc tcg gat ata gta act att ggg 192Pro Trp Gln Asp Thr Trp Glu Ser Val Ser Asp Ile Val Thr Ile Gly
50 55 60aca tac ctt ata caa ttc ttg cta gaa ccc ggt ata ggt gga att cct 240Thr Tyr Leu Ile Gln Phe Leu Leu Glu Pro Gly Ile Gly Gly Ile Pro65 70 75 80gta ata ttt tca ata ata aac aaa ctc att ccg tct tct ggt caa tct 288Val Ile Phe Ser Ile Ile Asn Lys Leu Ile Pro Ser Ser Gly Gln Ser85 90 95gtg gca gca ctt tct ata tgt gat tta gta tct ata att cgt aaa gag 336Val Ala Ala Leu Ser Ile Cys Asp Leu Val Ser Ile Ile Arg Lys Glu100105 110gta gac gag agc gtg tta agt gac ggg gtt gca gat ttt gag ggt gaa 384Val Asp Glu Ser Val Leu Ser Asp Gly Val Ala Asp Phe Glu Gly Glu115 120 125atg act gct tat caa gat tat tat ctt cat tat ctt gag gat tgg ctt 432Met Thr Ala Tyr Gln Asp Tyr Tyr Leu His Tyr Leu Glu Asp Trp Leu130 135 140aca gat aaa tca aat cct aaa aaa ctt gct gac gta gtt aaa cag ttc 480Thr Asp Lys Ser Asn Pro Lys Lys Leu Ala Asp Val Val Lys Gln Phe145 150 155 160caa gca cgg gaa gaa gat ttc act aaa ctt tta gca ggg tca tta tca 528Gln Ala Arg Glu Glu Asp Phe Thr Lys Leu Leu Ala Gly Ser Leu Ser165 170 175aga cag aaa gct gaa ata tta tta ttg cct acg tat gtg caa gct gca 576Arg Gln Lys Ala Glu Ile Leu Leu Leu Pro Thr Tyr Val Gln Ala Ala180 185 190aat gtg cat tta tta cta tta agg gac gca gtt aaa tat aaa aaa gaa 624Asn Val His Leu Leu Leu Leu Arg Asp Ala Val Lys Tyr Lys Lys Glu195 200 205tgg gga cta gtg tgt cca ccg ttg tat cca ggg tca ggg aga act gat 672Trp Gly Leu Val Cys Pro Pro Leu Tyr Pro Gly Ser Gly Arg Thr Asp210 215 220tgt aac gag cgg tta aaa gcg aaa ata aaa gag tat act aat tat tgt 720Cys Asn Glu Arg Leu Lys Ala Lys Ile Lys Glu Tyr Thr Asn Tyr Cys225 230 235 240gta ggg tgg tat aac aag ggt tta gat cag ata aga cag gcg ggt aca 768Val Gly Trp Tyr Asn Lys Gly Leu Asp Gln Ile Arg Gln Ala Gly Thr245 250 255agt gct gaa gtt tgg tcg aaa ttt aat aaa ttt cgt aga gaa atg acg 816Ser Ala Glu Val Trp Sar Lys Phe Asn Lys Phe Arg Arg Glu Met Thr260 265 270ttg gcg gta ttg gat att att gct ata ttt cca act tat gat ttt gaa 864Leu Ala Val Leu Asp Ile Ile Ala Ile Phe Pro Thr Tyr Asp Phe Glu275 280 285aaa tat cca tta gca aca agt gta gag tta act agg gaa att tat aca 912Lys Tyr Pro Leu Ala Thr Ser Val Glu Leu Thr Arg Glu Ile Tyr Thr290 295 300gat cca gtg gga tat tca ggg gga aat tat ggt tgg gaa cgg ttt ttt 960Asp Pro Val Gly Tyr ser Gly Gly Asn Tyr Gly Trp Glu Arg Phe Phe305 310 315 320agc ttt aat tcg gta gaa gca aat gga aca cgg gga cct ggt tta gtt 1008Ser Phe Asn Ser Val Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu Val
325 330 335act tgg ctt caa gct ata gat ata tat agt cat tct att aat ctt cag 1056Thr Trp Leu Gln Ala Ile Asp Ile Tyr Ser His Ser Ile Asn Leu Gln340 345 350ctt ggt tat ctt agt ggc tgg ggg gga act cgt cat tat gaa gac ttc 1104Leu Gly Tyr Leu Ser Gly Trp Gly Gly Thr Arg His Tyr Glu Asp Phe355 360 365aca aag ggt aac ggt gct ttt caa cgt atg tct gga act acg agt aat 1152Thr Lys Gly Asn Gly Ala Phe Gln Arg Met Ser Gly Thr Thr Ser Asn370 375 380aat cca cgt aat att att ttt ggc aat acc gat ata ttt aaa att att 1200Asn Pro Arg Asn Ile Ile Phe Gly Asn Thr Asp Ile Phe Lys Ile Ile385 390 395 400tca tta gct aga tat gca atg caa ccg ttt gtt ggg tat tca atc cca 1248Ser Leu Ala Arg Tyr Ala Met Gln Pro Phe Val Gly Tyr Ser Ile Pro405 410 415cgg cat ctt gtt tca cgt gca gaa ttt ttt ccg aca aca cta aat act 1296Arg His Leu Val Ser Arg Ala Glu Phe Phe Pro Thr Thr Leu Asn Thr420 425 430ttc ctg tat gag gta aac agt tct ggg tac tca cag aca att gaa tct 1344Phe Leu Tyr Glu Val Asn Ser Ser Gly Tyr Ser Gln Thr Ile Glu Ser435 440 445gtg tta cca ggt att aat aag gat cta cca cct agt cgt aca aat tac 1392Val Leu Pro Gly Ile Asn Lys Asp Leu Pro Pro Ser Arg Thr Asn Tyr450 455 460tct cat aga tta tca aat gcg gca tgt gtt caa aat gaa acc tcc aga 1440Ser His Arg Leu Ser Asn Ala Ala Cys Val Gln Asn Glu Thr Ser Arg465 470 475 480gtt aac gta ttt ggt tgg aca cat aca agt atg aaa aaa gat aat cga 1488Val Asn Val Phe Gly Trp Thr His Thr Ser Met Lys Lys Asp Asn Arg485 490 495att tat cca gat aaa att acg caa att cct gca gta aaa gct ttt gcc 1536Ile Tyr Pro Asp Lys Ile Thr Gln Ile Pro Ala Val Lys Ala Phe Ala500 505 510cta cca gca ggt aca gga tat gca gga ggt tac gtc aca gct ggg cct 1584Leu Pro Ala Gly Thr Gly Tyr Ala Gly Gly Tyr Val Thr Ala Gly Pro515 520 525ggt tat aca gga gga gat gta gta acg tta cct tat caa gca agt tta 1632Gly Tyr Thr Gly Gly Asp Val Val Thr Leu Pro Tyr Gln Ala Ser Leu530 535 540aaa ata cgt tta act tct gca ccc acg aat aaa aat tac cgt gtt aga 1680Lys Ile Arg Leu Thr Ser Ala Pro Thr Asn Lys Asn Tyr Arg Val Arg545 550 555 560ctt cgc tac gcg agt gga gga cct ggt ccg ttc cga gta gaa aga tgg 1728Leu Arg Tyr Ala Ser Gly Gly Pro Gly Pro Phe Arg Val Glu Arg Trp565 570 575tcg cca agt tct gtt tca aat gct aat ttt tct cgt cca gct aca ggt 1776Ser Pro Ser Ser Val Ser Asn Ala Asn Phe Ser Arg Pro Ala Thr Gly580 585 590ggc tat agt tca ttt gat tat gtg gac acc tta gtt act aca ttt aat 1824Gly Tyr Ser Ser Phe Asp Tyr Val Asp Thr Leu Val Thr Thr Phe Asn
595 600 605caa tca ggt gtt gaa ata att ata caa aat cta tct ggt tac cac ctt 1872Gln Ser Gly Val Glu Ile Ile Ile Gln Asn Leu Ser Gly Tyr His Leu610 615 620att gtt gac aaa gtc gaa ttt atc cca att gac atc caa att gaa aaa 1920Ile Val Asp Lys Val Glu Phe Ile Pro Ile Asp Ile Gln Ile Glu Lys625 630 635 640tgt acg aaa tgt caa ttc gaa gga gac ata tgt aga tgt gaa gga gta 1968Cys Thr Lys Cys Gln Phe Glu Gly Asp Ile Cys Arg Cys Glu Gly Val645 650 655caa tcc ttg gaa aca aaa aaa gag att gta aat agt tta ttt atc aat 2016Gln Ser Leu Glu Thr Lys Lys Glu Ile Val Asn Ser Leu Phe Ile Asn660 665 670taa 2019*<210>27<211>672<212>PRT<213>Bacillus thuringiensis<400>27Met Lys Asn Met Asn Ser Tyr Gln Asn Thr Asn Glu Tyr Glu Ile Leu1 5 10 15Asp Gly Ser Pro Asn Asn Thr Asn Met Ser Asn Arg Tyr Pro Phe Ala20 25 30Lys Asp Pro Asn Ile Phe Pro Ile Asn Leu Asp Ala Cys Gln Gly Arg35 40 45Pro Trp Gln Asp Thr Trp Glu Ser Val Ser Asp Ile Val Thr Ile Gly50 55 60Thr Tyr Leu Ile Gln Phe Leu Leu Glu Pro Gly Ile Gly Gly Ile Pro65 70 75 80Val Ile Phe Ser Ile Ile Asn Lys Leu Ile Pro Ser Ser Gly Gln Ser85 90 95Val Ala Ala Leu Ser Ile Cys Asp Leu Val Ser Ile Ile Arg Lys Glu100 105 110Val Asp Glu Ser Val Leu Ser Asp Gly Val Ala Asp Phe Glu Gly Glu115 120 125Met Thr Ala Tyr Gln Asp Tyr Tyr Leu His Tyr Leu Glu Asp Trp Leu130 135 140Thr Asp Lys Ser Asn Pro Lys Lys Leu Ala Asp Val Val Lys Gln Phe145 150 155 160Gln Ala Arg Glu Glu Asp Phe Thr Lys Leu Leu Ala Gly Ser Leu Ser165 170 175Arg Gln Lys Ala Glu Ile Leu Leu Leu Pro Thr Tyr Val Gln Ala Ala180 185 190Asn Val His Leu Leu Leu Leu Arg Asp Ala Val Lys Tyr Lys Lys Glu195 200 205Trp Gly Leu Val Cys Pro Pro Leu Tyr Pro Gly Ser Gly Arg Thr Asp210 215 220Cys Asn Glu Arg Leu Lys Ala Lys Ile Lys Glu Tyr Thr Asn Tyr Cys225 230 235 240Val Gly Trp Tyr Asn Lys Gly Leu Asp Gln Ile Arg Gln Ala Gly Thr245 250 255Ser Ala Glu Val Trp Ser Lys Phe Asn Lys Phe Arg Arg Glu Met Thr260 265 270Leu Ala Val Leu Asp Ile Ile Ala Ile Phe Pro Thr Tyr Asp Phe Glu275 280 285Lys Tyr Pro Leu Ala Thr Ser Val Glu Leu Thr Arg Glu Ile Tyr Thr290 295 300Asp Pro Val Gly Tyr Ser Gly Gly Asn Tyr Gly Trp Glu Arg Phe Phe
305 310 315 320Ser Phe Asn Ser Val Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu Val325 330 335Thr Trp Leu Gln Ala Ile Asp Ile Tyr Ser His Ser Ile Asn Leu Gln340 345 350Leu Gly Tyr Leu Ser Gly Trp Gly Gly Thr Arg His Tyr Glu Asp Phe355 360 365Thr Lys Gly Asn Gly Ala Phe Gln Arg Met Ser Gly Thr Thr Ser Asn370 375 380Asn Pro Arg Asn Ile Ile Phe Gly Asn Thr Asp Ile Phe Lys Ile Ile385 390 395 400Ser Leu Ala Arg Tyr Ala Met Gln Pro Phe Val Gly Tyr Ser Ile Pro405 410 415Arg His Leu Val Ser Arg Ala Glu Phe Phe Pro Thr Thr Leu Asn Thr420 425 430Phe Leu Tyr Glu Val Asn Ser Ser Gly Tyr Ser Gln Thr Ile Glu Ser435 440 445Val Leu Pro Gly Ile Asn Lys Asp Leu Pro Pro Ser Arg Thr Asn Tyr450 455 460Ser His Arg Leu Ser Asn Ala Ala Cys Val Gln Asn Glu Thr Ser Arg465 470 475 480Val Asn Val Phe Gly Trp Thr His Thr Ser Met Lys Lys Asp Asn Arg485 490 495Ile Tyr Pro Asp Lys Ile Thr Gln Ile Pro Ala Val Lys Ala Phe Ala500 505 510Leu Pro Ala Gly Thr Gly Tyr Ala Gly Gly Tyr Val Thr Ala Gly Pro515 520 525Gly Tyr Thr Gly Gly Asp Val Val Thr Leu Pro Tyr Gln Ala Ser Leu530 535 540Lys Ile Arg Leu Thr Ser Ala Pro Thr Asn Lys Asn Tyr Arg Val Arg545 550 555 560Leu Arg Tyr Ala Ser Gly Gly Pro Gly Pro Phe Arg Val Glu Arg Trp565 570 575Ser Pro Ser Ser Val Ser Asn Ala Asn Phe Ser Arg Pro Ala Thr Gly580 585 590Gly Tyr Ser Ser Phe Asp Tyr Val Asp Thr Leu Val Thr Thr Phe Asn595 600 605Gln Ser Gly Val Glu Ile Ile Ile Gln Asn Leu Ser Gly Tyr His Leu610 615 620Ile Val Asp Lys Val Glu Phe Ile Pro Ile Asp Ile Gln Ile Glu Lys625 630 635 640Cys Thr Lys Cys Gln Phe Glu Gly Asp Ile Cys Arg Cys Glu Gly Val645 650 655Gln Ser Leu Glu Thr Lys Lys Glu Ile Val Asn Ser Leu Phe Ile Asn660 665 670<210>28<211>2010<212>DNA<213>Bacillus thuringiensis<220>
<221>CDS<222>(1)...(2010)<400>28atg aat tct tat caa aat aca aat gaa tat gaa att ctg gat ggt tcc 48Met Asn Ser Tyr Gln Asn Thr Asn Glu Tyr Glu Ile Leu Asp Gly Ser1 5 10 15ccg aat aac aca aat atg tca aac aga tat cct ttt gca aag gat cca 96Pro Asn Asn Thr Asn Met Ser Asn Arg Tyr Pro Phe Ala Lys Asp Pro20 25 30aat ata ttt cct att aac ctg gac gct tgt cag gga agg cca tgg caa 144Asn Ile Phe Pro Ile Asn Leu Asp Ala Cya Gln Gly Arg Pro Trp Gln35 40 45
gat acg tgg gaa tca gtc tcg gat ata gta act att ggg aca tac ctt 192Asp Thr Trp Glu Ser Val Ser Asp Ile Val Thr Ile Gly Thr Tyr Leu50 55 60ata caa ttc ttg cta gaa ccc ggt ata ggt gga att cct gta ata ttt 240Ile Gln Phe Leu Leu Glu Pro Gly Ile Gly Gly Ile Pro Val Ile Phe65 70 75 80tca ata ata aac aaa ctc att ccg tct tct ggt caa tct gtg gca gca 288Ser Ile Ile Asn Lys Leu Ile Pro Ser Ser Gly Gln Ser Val Ala Ala85 90 95ctt tct ata tgt gat tta gta tct ata att cgt aaa gag gta gac gag 336Leu Ser Ile Cys Asp Leu Val Ser Ile Ile Arg Lys Glu Val Asp Glu100 105 110agc gtg tta agt gac ggg gtt gca gat ttt gag ggt gaa atg act gct 384Ser Val Leu Ser Asp Gly Val Ala Asp Phe Glu Gly Glu Met Thr Ala115 120 125tat caa gat tat tat ctt cat tat ctt gag gat tgg ctt aca gat aaa 432Tyr Gln Asp Tyr Tyr Leu His Tyr Leu Glu Asp Trp Leu Thr Asp Lys130 135 140tca aat cct aaa aaa ctt gct gac gta gtt aaa cag ttc caa gca cgg 480Ser Asn Pro Lys Lys Leu Ala Asp Val Val Lys Gln Phe Gln Ala Arg145 150 155 160gaa gaa gat ttc act aaa ctt tta gca ggg tca tta tca aga cag aaa 528Glu Glu Asp Phe Thr Lys Leu Leu Ala Gly Ser Leu Ser Arg Gln Lys165 170 175gct gaa ata tta tta ttg cct acg tat gtg caa gct gca aat gtg cat 576Ala Glu Ile Leu Leu Leu Pro Thr Tyr Val Gln Ala Ala Asn Val His180 185 190tta tta cta tta agg gac gca gtt aaa tat aaa aaa gaa tgg gga cta 624Leu Leu Leu Leu Arg Asp Ala Val Lys Tyr Lys Lys Glu Trp Gly Leu195 200 205gtg tgt cca ccg ttg tat cca ggg tca ggg aga act gat tgt aac gag 672Val Cys Pro Pro Leu Tyr Pro Gly Ser Gly Arg Thr Asp Cys Asn Glu210 215 220cgg tta aaa gcg aaa ata aaa gag tat act aat tat tgt gta ggg tgg 720Arg Leu Lys Ala Lys Ile Lys Glu Tyr Thr Asn Tyr Cys Val Gly Trp225 230 235 240tat aac aag ggt tta gat cag ata aga cag gcg ggt aca agt gct gaa 768Tyr Asn Lys Gly Leu Asp Gln Ile Arg Gln Ala Gly Thr Ser Ala Glu245 250 255gtt tgg tcg aaa ttt aat aaa ttt cgt aga gaa atg acg ttg gcg gta 816Val Trp Ser Lys Phe Asn Lys Phe Arg Arg Glu Met Thr Leu Ala Val260 265 270ttg gat att att gct ata ttt cca act tat gat ttt gaa aaa tat cca 864Leu Asp Ile Ile Ala Ile Phe Pro Thr Tyr Asp Phe Glu Lys Tyr Pro275 280 285tta gca aca agt gta gag tta act agg gaa att tat aca gat cca gtg 912Leu Ala Thr Ser Val Glu Leu Thr Arg Glu Ile Tyr Thr Asp Pro Val290 295 300gga tat tca ggg gga aat tat ggt tgg gaa cgg ttt ttt agc ttt aat 960Gly Tyr Ser Gly Gly Asn Tyr Gly Trp Glu Arg Phe Phe Ser Phe Asn305 310 315 320
tcg gta gaa gca aat gga aca cgg gga cct ggt tta gtt act tgg ctt 1008Ser Val Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu Val Thr Trp Leu325 330 335caa gct ata gat ata tat agt cat tct att aat ctt cag ctt ggt tat 1056Gln Ala Ile Asp Ile Tyr Ser His Ser Ile Asn Leu Gln Leu Gly Tyr340 345 350ctt agt ggc tgg ggg gga act cgt cat tat gaa gac ttc aca aag ggt 1104Leu Ser Gly Trp Gly Gly Thr Arg His Tyr Glu Asp Phe Thr Lys Gly355 360 365aac ggt gct ttt caa cgt atg tct gga act acg agt aat aat cca cgt 1152Asn Gly Ala Phe Gln Arg Met Ser Gly Thr Thr Ser Asn Asn Pro Arg370 375 380aat att att ttt ggc aat acc gat ata ttt aaa att att tca tta gct 1200Asn Ile Ile Phe Gly Asn Thr Asp Ile Phe Lys Ile Ile Ser Leu Ala385 390 395 400aga tat gca atg caa ccg ttt gtt ggg tat tca atc cca cgg cat ctt 1248Arg Tyr Ala Met Gln Pro Phe Val Gly Tyr Ser Ile Pro Arg His Leu405 410 415gtt tca cgt gca gaa ttt ttt ccg aca aca cta aat act ttc ctg tat 1296Val Ser Arg Ala Glu Phe Phe Pro Thr Thr Leu Asn Thr Phe Leu Tyr420 425 430gag gta aac agt tct ggg tac tca cag aca att gaa tct gtg tta cca 1344Glu Val Asn Ser Ser Gly Tyr Ser Gln Thr Ile Glu Ser Val Leu Pro435 440 445ggt att aat aag gat cta cca cct agt cgt aca aat tac tct cat aga 1392Gly Ile Asn Lys Asp Leu Pro Pro Ser Arg Thr Asn Tyr Ser His Arg450 455 460tta tca aat gcg gca tgt gtt caa aat gaa acc tcc aga gtt aac gta 1440Leu Ser Asn Ala Ala Cys Val Gln Asn Glu Thr Ser Arg Val Asn Val465 470 475 480ttt ggt tgg aca cat aca agt atg aaa aaa gat aat cga att tat cca 1488Phe Gly Trp Thr His Thr Ser Met Lys Lys Asp Asn Arg Ile Tyr Pro485 490 495gat aaa att acg caa att cct gca gta aaa gct ttt gcc cta cca gca 1536Asp Lys Ile Thr Gln Ile Pro Ala Val Lys Ala Phe Ala Leu Pro Ala500 505 510ggt aca gga tat gca gga ggt tac gtc aca gct ggg cct ggt tat aca 1584Gly Thr Gly Tyr Ala Gly Gly Tyr Val Thr Ala Gly Pro Gly Tyr Thr515 520 525gga gga gat gta gta acg tta cct tat caa gca agt tta aaa ata cgt 1632Gly Gly Asp Val Val Thr Leu Pro Tyr Gln Ala Ser Leu Lys Ile Arg530 535 540tta act tct gca ccc acg aat aaa aat tac cgt gtt aga ctt cgc tac 1680Leu Thr Ser Ala Pro Thr Asn Lys Asn Tyr Arg Val Arg Leu Arg Tyr545 550 555 560gcg agt gga gga cct ggt ccg ttc cga gta gaa aga tgg tcg cca agt 1728Ala Ser Gly Gly Pro Gly Pro Phe Arg Val Glu Arg Trp Ser Pro Ser565 570 575tct gtt tca aat gct aat ttt tct cgt cca gct aca ggt ggc tat agt 1776Ser Val Ser Asn Ala Asn Phe Ser Arg Pro Ala Thr Gly Gly Tyr Ser580 585 590
tca ttt gat tat gtg gac acc tta gtt act aca ttt aat caa tca ggt 1824Ser Phe Asp Tyr Val Asp Thr Leu Val Thr Thr Phe Asn Gln Ser Gly595 600 605gtt gaa ata att ata caa aat cta tct ggt tac cac ctt att gtt gac 1872Val Glu Ile Ile Ile Gln Asn Leu Ser Gly Tyr His Leu Ile Val Asp610 615 620aaa gtc gaa ttt atc cca att gac atc caa att gaa aaa tgt acg aaa 1920Lys Val Glu Phe Ile Pro Ile Asp Ile Gln Ile Glu Lys Cys Thr Lys625 630 635 640tgt caa ttc gaa gga gac ata tgt aga tgt gaa gga gta caa tcc ttg 1968Cys Gln Phe Glu Gly Asp Ile Cys Arg Cys Glu Gly Val Gln Ser Leu645 650 655gaa aca aaa aaa gag att gta aat agt tta ttt atc aat taa 2010Glu Thr Lys Lys Glu Ile Val Asn Ser Leu Phe Ile Asn *660 665<210>29<211>669<212>PRT<213>Bacillus thuringiensis<400>29Met Asn Ser Tyr Gln Asn Thr Asn Glu Tyr Glu Ile Leu Asp Gly Ser1 5 10 15Pro Asn Asn Thr Asn Met Ser Asn Arg Tyr Pro Phe Ala Lys Asp Pro20 25 30Asn Ile Phe Pro Ile Asn Leu Asp Ala Cys Gln Gly Arg Pro Trp Gln35 40 45Asp Thr Trp Glu Ser Val Ser Asp Ile Val Thr Ile Gly Thr Tyr Leu50 55 60Ile Gln Phe Leu Leu Glu Pro Gly Ile Gly Gly Ile Pro Val Ile Phe65 70 75 80Ser Ile Ile Asn Lys Leu Ile Pro Ser Ser Gly Gln Ser Val Ala Ala85 90 95Leu Ser Ile Cys Asp Leu Val Ser Ile Ile Arg Lys Glu Val Asp Glu100 105 110Ser Val Leu Ser Asp Gly Val Ala Asp Phe Glu Gly Glu Met Thr Ala115 120 125Tyr Gln Asp Tyr Tyr Leu His Tyr Leu Glu Asp Trp Leu Thr Asp Lys130 135 140Ser Asn Pro Lys Lys Leu Ala Asp Val Val Lys Gln Phe Gln Ala Arg145 150 155 160Glu Glu Asp Phe Thr Lys Leu Leu Ala Gly Ser Leu Ser Arg Gln Lys165 170 175Ala Glu Ile Leu Leu Leu Pro Thr Tyr Val Gln Ala Ala Asn Val His180 185 190Leu Leu Leu Leu Arg Asp Ala Val Lys Tyr Lys Lys Glu Trp Gly Leu195 200 205Val Cys Pro Pro Leu Tyr Pro Gly Ser Gly Arg Thr Asp Cys Asn Glu210 215 220Arg Leu Lys Ala Lys Ile Lys Glu Tyr Thr Asn Tyr Cys Val Gly Trp225 230 235 240Tyr Asn Lys Gly Leu Asp Gln Ile Arg Gln Ala Gly Thr Ser Ala Glu245 250 255Val Trp Ser Lys Phe Asn Lys Phe Arg Arg Glu Met Thr Leu Ala Val260 265 270Leu Asp Ile Ile Ala Ile Phe Pro Thr Tyr Asp Phe Glu Lys Tyr Pro275 280 285Leu Ala Thr Ser Val Glu Leu Thr Arg Glu Ile Tyr Thr Asp Pro Val290 295 300Gly Tyr Ser Gly Gly Asn Tyr Gly Trp Glu Arg Phe Phe Ser Phe Asn
305 310 315 320Ser Val Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu Val Thr Trp Leu325 330 335Gln Ala Ile Asp Ile Tyr Ser His Ser Ile Asn Leu Gln Leu Gly Tyr340 345 350Leu Ser Gly Trp Gly Gly Thr Arg His Tyr Glu Asp Phe Thr Lys Gly355 360 365Asn Gly Ala Phe Gln Arg Met Ser Gly Thr Thr Ser Asn Asn Pro Arg370 375 380Asn Ile Ile Phe Gly Asn Thr Asp Ile Phe Lys Ile Ile Ser Leu Ala385 390 395 400Arg Tyr Ala Met Gln Pro Phe Val Gly Tyr Ser Ile Pro Arg His Leu405 410 415Val Ser Arg Ala Glu Phe Phe Pro Thr Thr Leu Asn Thr Phe Leu Tyr420 425 430Glu Val Asn Ser Ser Gly Tyr Ser Gln Thr Ile Glu Ser Val Leu Pro435 440 445Gly Ile Asn Lys Asp Leu Pro Pro Ser Arg Thr Asn Tyr Ser His Arg450 455 460Leu Ser Asn Ala Ala Cys Val Gln Asn Glu Thr Ser Arg Val Asn Val465 470 475 480Phe Gly Trp Thr His Thr Ser Met Lys Lys Asp Asn Arg Ile Tyr Pro485 490 495Asp Lys Ile Thr Gln Ile Pro Ala Val Lys Ala Phe Ala Leu Pro Ala500 505 510Gly Thr Gly Tyr Ala Gly Gly Tyr Val Thr Ala Gly Pro Gly Tyr Thr515 520 525Gly Gly Asp Val Val Thr Leu Pro Tyr Gln Ala Ser Leu Lys Ile Arg530 535 540Leu Thr Ser Ala Pro Thr Asn Lys Asn Tyr Arg Val Arg Leu Arg Tyr545 550 555 560Ala Ser Gly Gly Pro Gly Pro Phe Arg Val Glu Arg Trp Ser Pro Ser565 570 575Ser Val Ser Asn Ala Asn Phe Ser Arg Pro Ala Thr Gly Gly Tyr Ser580 585 590Ser Phe Asp Tyr Val Asp Thr Leu Val Thr Thr Phe Asn Gln Ser Gly595 600 605Val Glu Ile Ile Ile Gln Asn Leu Ser Gly Tyr His Leu Ile Val Asp610 615 620Lys Val Glu Phe Ile Pro Ile Asp Ile Gln Ile Glu Lys Cys Thr Lys625 630 635 640Cys Gln Phe Glu Gly Asp Ile Cys Arg Cys Glu Gly Val Gln Ser Leu645 650 655Glu Thr Lys Lys Glu Ile Val Asn Ser Leu Phe Ile Asn660 665<210>30<211>1176<212>PRT<213>Bacillus thuringiensi<400>30Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu1 5 10 15Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly20 25 30Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser35 40 45Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile50 55 60Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Pro Val Gln Ile65 70 75 80Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala85 90 95Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu100 105 110Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu
115 120 125Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala130 135 140Ile Pro Leu Leu Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val145 150 155 160Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser165 170 175Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg180 185 190Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp Tyr Ala Val195 200 205Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg210 215 220Asp Trp Val Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val225 230 235 240Leu Asp Ile Val Ala Leu Phe Ser Asn Tyr Asp Ser Arg Arg Tyr Pro245 250 255Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val260 265 270Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Met Ala Gln Arg Ile Glu275 280 285Gln Asn Ile Arg Gln Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr290 295 300Ile Tyr Thr Asp Val His Arg Gly Phe Asn Tyr Trp Ser Gly His Gln305 310 315 320Ile Thr Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Ala Phe Pro325 330 335Leu Phe Gly Asn Ala Gly Asn Ala Ala Pro Pro Val Leu Val Ser Leu340 345 350Thr Gly Leu Gly Ile Phe Arg Thr Leu Ser Ser Pro Leu Tyr Arg Arg355 360 365Ile Ile Leu Gly Ser Gly Pro Asn Asn Gln Glu Leu Phe Val Leu Asp370 375 380Gly Thr Glu Phe Ser Phe Ala Ser Leu Thr Thr Asn Leu Pro Ser Thr385 390 395 400Ile Tyr Arg Gln Arg Gly Thr Val Asp Ser Leu Asp Val Ile Pro Pro405 410 415Gln Asp Asn Ser Val Pro Pro Arg Ala Gly Phe Ser His Arg Leu Ser420 425 430His Val Thr Met Leu Ser Gln Ala Ala Gly Ala Val Tyr Thr Leu Arg435 440 445Ala Pro Thr Phe Ser Trp Gln His Arg Ser Ala Glu Phe Asn Asn Ile450 455 460Ile Pro Ser Ser Gln Ile Thr Gln Ile Pro Leu Thr Lys Ser Thr Asn465 470 475 480Leu Gly Ser Gly Thr Ser Val Val Lys Gly Pro Gly Phe Thr Gly Gly485 490 495Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln Ile Ser Thr Leu Arg Val500 505 510Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr Arg Val Arg Ile Arg Tyr515 520 525Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser Ile Asp Gly Arg Pro530 535 540Ile Asn Gln Gly Asn Phe Ser Ala Thr Met Ser Ser Gly Ser Asn Leu545 550 555 560Gln Ser Gly Ser Phe Arg Thr Val Gly Phe Thr Thr Pro Phe Asn Phe565 570 575Ser Asn Gly Ser Ser Val Phe Thr Leu Ser Ala His Val Phe Asn Ser580 585 590Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu Phe Val Pro Ala Glu Val595 600 605Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg Ala Gln Lys Ala Val Asn610 615 620Glu Leu Phe Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr625 630 635 640Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Glu Cys Leu Ser Asp645 650 655Glu Phe Cys Leu Asp Glu Lys Gln Glu Leu Ser Glu Lys Val Lys His
660 665 670Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe675 680 685Arg Gly Ile Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr Asp690 695 700Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr705 710 715 720Leu Leu Gly Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys725 730 735Ile Asp Glu Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly740 745 750Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn755 760 765Ala Lys His Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro770 775 780Leu Ser Ala Gln Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys785 790 795 800Ala Pro His Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp805 810 815Gly Glu Lys Cys Ala His His Ser His His Phe Ser Leu Asp Ile Asp820 825 830Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe835 840 845Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe850 855 860Leu Glu Glu Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val Lys Arg865 870 875 880Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr885 890 895Asn Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val900 905 910Asn Ser Gln Tyr Asp Gln Leu Gln Ala Asp Thr Asn Ile Ala Met Ile915 920 925His Ala Ala Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr Leu Pro930 935 940Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu945 950 955 960Glu Gly Arg Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val965 970 975Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn Val Lys980 985 990Gly His Val Asp Val Glu Glu Gln Asn Asn Gln Arg Ser Val Leu Val995 10001005Val Pro Glu Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro101010151020Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly102510301035 1040Glu Gly Cys Val Thr Ile His Glu Ile Glu Asn Asn Thr Asp Glu Leu104510501055Lys Phe Ser Asn Cys Val Glu Glu Glu Ile Tyr Pro Asn Asn Thr Val106010651070Thr Cys Asn Asp Tyr Thr Val Asn Gln Glu Glu Tyr Gly Gly Ala Tyr107510801085Thr Ser Arg Asn Arg Gly Tyr Asn Glu Ala Pro Ser Val Pro Ala Asp109010951100Tyr Ala Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg Arg Glu110511101115 1120Asn Pro Cys Glu Phe Asn Arg Gly Tyr Arg Asp Tyr Thr Pro Leu Pro112511301135Val Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp Lys114011451150Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp Ser115511601165Val Glu Leu Leu Leu Met Glu Glu11701175<210>31
<211>1178<212>PRT<213>Bacillus thuringiensi<400>31Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu1 5 10 15Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly20 25 30Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser35 40 45Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile50 55 60Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile65 70 75 80Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala85 90 95Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu100 105 110Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu115 120 125Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala130 135 140Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val145 150 155 160Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser165 170 175Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg180 185 190Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp Tyr Ala Val195 200 205Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg210 215 220Asp Trp Val Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val225 230 235 240Leu Asp Ile Val Ala Leu Phe Pro Asn Tyr Asp Ser Arg Arg Tyr Pro245 250 255Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val260 265 270Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu275 280 285Arg Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr290 295 300Ile Tyr Thr Asp Ala His Arg Gly Tyr Tyr Tyr Trp Ser Gly His Gln305 310 315 320Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro325 330 335Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala340 345 350Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg355 360 365Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp370 375 380Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val385 390 395 400Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln405 410 415Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His420 425 430Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile435 440 445Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Glu Phe Asn Asn450 455 460Ile Ile Ala Ser Asp Ser Ile Thr Gln Ile Pro Ala Val Lys Gly Asn465 470 475 480Phe Leu Phe Asn Gly Ser Val Ile Ser Gly Pro Gly Phe Thr Gly Gly485 490 495Asp Leu Val Arg Leu Asn Ser Ser Gly Asn Asn Ile Gln Asn Arg Gly
500 505 510Tyr Ile Glu Val Pro Ile His Phe Pro Ser Thr Ser Thr Arg Tyr Arg515 520 525Val Arg Val Arg Tyr Ala Ser Val Thr Pro Ile His Leu Asn Val Asn530 535 540Trp Gly Asn Ser Ser Ile Phe Ser Asn Thr Val Pro Ala Thr Ala Thr545 550 555 560Ser Leu Asp Asn Leu Gln Ser Ser Asp Phe Gly Tyr Phe Glu Ser Ala565 570 575Asn Ala Phe Thr Ser Ser Leu Gly Asn Ile Val Gly Val Arg Asn Phe580 585 590Ser Gly Thr Ala Gly Val Ile Ile Asp Arg Phe Glu Phe Ile Pro Val595 600 605Thr Ala Thr Leu Glu Ala Glu Tyr Asn Leu Glu Arg Ala Gln Lys Ala610 615 620Val Asn Ala Leu Phe Thr Ser Thr Asn Gln Leu Gly Leu Lys Thr Asn625 630 635 640Val Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Thr Tyr Leu645 650 655Ser Asp Glu Phe Cys Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val660 665 670Lys His Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Ser675 680 685Asn Phe Lys Asp Ile Asn Arg Gln Pro Glu Arg Gly Trp Gly Gly Ser690 695 700Thr Gly Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr705 710 715 720Val Thr Leu Ser Gly Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr725 730 735Gln Lys Ile Asp Glu Ser Lys Leu Lys Ala Phe Thr Arg Tyr Gln Leu740 745 750Arg Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg755 760 765Tyr Asn Ala Lys His Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu770 775 780Trp Pro Leu Ser Ala Gln Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn785 790 795 800Arg Cys Ala Pro His Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys805 810 815Arg Asp Gly Glu Lys Cys Ala His His Ser His His Phe Ser Leu Asp820 825 830Ile Asp Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val835 840 845Ile Phe Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu850 855 860Glu Phe Leu Glu Glu Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val865 870 875 880Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp885 890 895Glu Thr Asn Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu900 905 910Phe Val Asn Ser Gln Tyr Asp Gln Leu Gln Ala Asp Thr Asn Ile Ala915 920 925Met Ile His Ala Ala Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr930 935 940Leu Pro Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu945 950 955 960Glu Leu Glu Gly Arg Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg965 970 975Asn Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn980 985 990Val Lys Gly His Val Asp Val Glu Glu Gln Asn Asn Gln Arg Ser Val995 10001005Leu Val Val Pro Glu Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val101010151020Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly102510301035 1040Tyr Gly Glu Gly Cys Val Thr Ile His Glu Ile Glu Asn Asn Thr Asp
104510501055Glu Leu Lys Phe Ser Asn Cys Val Glu Glu Glu Ile Tyr Pro Asn Asn106010651070Thr Val Thr Cys Asn Asp Tyr Thr Val Asn Gln Glu Glu Tyr Gly Gly107510801085Ala Tyr Thr Ser Arg Asn Arg Gly Tyr Asn Glu Ala Pro Ser Val Pro109010951100Ala Asp Tyr Ala Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg110511101115 1120Arg Glu Asn Pro Cys Glu Phe Asn Arg Gly Tyr Arg Asp Tyr Thr Pro112511301135Leu Pro Val Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr114011451150Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val115511601165Asp Ser Val Glu Leu Leu Leu Met Glu Glu11701175<210>32<211>1189<212>PRT<213>Bacillus thuringiensi<400>32Met Glu Glu Asn Asn Gln Asn Gln Cys Ile Pro Tyr Asn Cys Leu Ser1 5 10 15Asn Pro Glu Glu Val Leu Leu Asp Gly Glu Arg Ile Ser Thr Gly Asn20 25 30Ser Ser Ile Asp Ile Ser Leu Ser Leu Val Gln Phe Leu Val Ser Asn35 40 45Phe Val Pro Gly Gly Gly Phe Leu Val Gly Leu Ile Asp Phe Val Trp50 55 60Gly Ile Val Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile Glu65 70 75 80Gln Leu Ile Asn Glu Arg Ile Ala Glu Phe Ala Arg Asn Ala Ala Ile85 90 95Ala Asn Leu Glu Gly Leu Gly Asn Asn Phe Asn Ile Tyr Val Glu Ala100 105 110Phe Lys Glu Trp Glu Glu Asp Pro Asn Asn Pro Ala Thr Arg Thr Arg115 120 125Val Ile Asp Arg Phe Arg Ile Leu Asp Gly Leu Leu Glu Arg Asp Ile130 135 140Pro Ser Phe Arg Ile Ser Gly Phe Glu Val Pro Leu Leu Ser Val Tyr145 150 155 160Ala Gln Ala Ala Asn Leu His Leu Ala Ile Leu Arg Asp Ser Val Ile165 170 175Phe Gly Glu Arg Trp Gly Leu Thr Thr Ile Asn Val Asn Glu Asn Tyr180 185 190Asn Arg Leu Ile Arg His Ile Asp Glu Tyr Ala Asp His Cys Ala Asn195 200 205Thr Tyr Asn Arg Gly Leu Asn Asn Leu Pro Lys Ser Thr Tyr Gln Asp210 215 220Trp Ile Thr Tyr Asn Arg Leu Arg Arg Asp Leu Thr Leu Thr Val Leu225 230 235 240Asp Ile Ala Ala Phe Phe Pro Asn Tyr Asp Asn Arg Arg Tyr Pro Ile245 250 255Gln Pro Val Gly Gln Leu Thr Arg Glu Val Tyr Thr Asp Pro Leu Ile260 265 270Asn Phe Asn Pro Gln Leu Gln Ser Val Ala Gln Leu Pro Thr Phe Asn275 280 285Val Met Glu Ser Ser Ala Ile Arg Asn Pro His Leu Phe Asp Ile Leu290 295 300Asn Asn Leu Thr Ile Phe Thr Asp Trp Phe Ser Val Gly Arg Asn Phe305 310 315 320Tyr Trp Gly Gly His Arg Val Ile Ser Ser Leu Ile Gly Gly Gly Asn325 330 335Ile Thr Ser Pro Ile Tyr Gly Arg Glu Ala Asn Gln Glu Pro Pro Arg
340 345 350Ser Phe Thr Phe Asn Gly Pro Val Phe Arg Thr Leu Ser Asn Pro Thr355 360 365Leu Arg Leu Leu Gln Gln Pro Trp Pro Ala Pro Pro Phe Asn Leu Arg370 375 380Gly Val Glu Gly Val Glu Phe Ser Thr Pro Thr Asn Ser Phe Thr Tyr385 390 395 400Arg Gly Arg Gly Thr Val Asp Ser Leu Thr Glu Leu Pro Pro Glu Asp405 410 415Asn Ser Val Pro Pro Arg Glu Gly Tyr Ser His Arg Leu Cys His Ala420 425 430Thr Phe Val Gln Arg Ser Gly Thr Pro Phe Leu Thr Thr Gly Val Val435 440 445Phe Ser Trp Thr His Arg Ser Ala Thr Leu Thr Asn Thr Ile Asp Pro450 455 460Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe Arg Val Trp Gly465 470 475 480Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly Gly Asp Ile Leu485 490 495Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln Val Asn Ile Asn500 505 510Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg Tyr Ala Ser Ser515 520 525Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala Ser Thr Gly Val530 535 540Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys Thr Met Glu Ile545 550 555 560Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr Asp Phe Ser Asn565 570 575Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly Ile Ser Glu Gln580 585 590Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu Leu Tyr Ile Asp595 600 605Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu Ala Glu Ser Asp610 615 620Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe Thr Ser Ser Asn625 630 635 640Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His Ile Asp Gln Val645 650 655Ser Asn Leu Val Asp Cys Leu Ser Asp Glu Phe Cys Leu Asp Glu Lys660 665 670Arg Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg Leu Ser Asp Glu675 680 685Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile Asn Arg Gln Pro690 695 700Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile Gln Gly Gly Asp705 710 715 720Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Pro Gly Thr Val Asp Glu725 730 735Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu Ser Lys Leu Lys740 745 750Ala Tyr Thr Arg Tyr Glu Leu Arg Gly Tyr Ile Glu Asp Ser Gln Asp755 760 765Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His Glu Ile Val Asn770 775 780Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala Gln Ser Pro Ile785 790 795 800Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His Leu Glu Trp Asn805 810 815Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys Cys Ala His His820 825 830Ser His His Phe Thr Leu Asp Ile Asp Val Gly Cys Thr Asp Leu Asn835 840 845Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys Thr Gln Asp Gly850 855 860His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu Lys Pro Leu Leu865 870 875 880Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys Lys Trp Arg Asp
885 890 895Lys Arg Glu Lys Leu Gln Leu Glu Thr Asn Ile Val Tyr Lys Glu Ala900 905 910Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln Tyr Asp Arg Leu915 920 925Gln Val Asp Thr Asn Ile Ala Met Ile His Ala Ala Asp Lys Arg Val930 935 940His Arg Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser Val Ile Pro Gly945 950 955 960Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg Ile Phe Thr Ala965 970 975Tyr Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn Gly Asp Phe Asn980 985 990Asn Gly Leu Leu Cys Trp Asn Val Lys Gly His Val Asp Val Glu Glu995 10001005Gln Asn Asn His Arg Ser Val Leu Val Ile Pro Glu Trp Glu Ala Glu101010151020Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg102510301035 1040Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His104510501055Glu Ile Glu Asp Asn Thr Asp Glu Leu Lys Phe Ser Asn Cys Val Glu106010651070Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn Asn Tyr Thr Gly107510801085Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg Asn Gln Gly Tyr109010951100Asp Glu Ala Tyr Gly Asn Asn Pro Ser Val Pro Ala Asp Tyr Ala Ser110511101115 1120Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg Arg Glu Asn Pro Cys112511301135Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu Pro Ala Gly Tyr114011451150Val Thr Lys Asp Leu Glu Tyr Phe Pro Glu Thr Asp Lys Val Trp Ile115511601165Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp Ser Val Glu Leu117011751180Leu Leu Met Glu Glu1185<210>33<211>719<212>PRT<213>Bacillus thuringiensi<400>33Met Lys Leu Lys Asn Gln Asp Lys His Gln Ser Phe Ser Ser Asn Ala1 5 10 15Lys Val Asp Lys Ile Ser Thr Asp Ser Leu Lys Asn Glu Thr Asp Ile20 25 30Glu Leu Gln Asn Ile Asn His Glu Asp Cys Leu Lys Met Ser Glu Tyr35 40 45Glu Asn Val Glu Pro Phe Val Ser Ala Ser Thr Ile Gln Thr Gly Ile50 55 60Gly Ile Ala Gly Lys Ile Leu Gly Thr Leu Gly Val Pro Phe Ala Gly65 70 75 80Gln Val Ala Ser Leu Tyr Ser Phe Ile Leu Gly Glu Leu Trp Pro Lys85 90 95Gly Lys Asn Gln Trp Glu Ile Phe Met Glu His Val Glu Glu Ile Ile100 105 110Asn Gln Lys Ile Ser Thr Tyr Ala Arg Asn Lys Ala Leu Thr Asp Leu115 120 125Lys Gly Leu Gly Asp Ala Leu Ala Val Tyr His Asp Ser Leu Glu Ser130 135 140Trp Val Gly Asn Arg Asn Asn Thr Arg Ala Arg Ser Val Val Lys Ser145 150 155 160Gln Tyr Ile Ala Leu Glu Leu Met Phe Val Gln Lys Leu Pro Ser Phe
165 170 175Ala Val Ser Gly Glu Glu Val Pro Leu Leu Pro Ile Tyr Ala Gln Ala180 185 190Ala Asn Leu His Leu Leu Leu Leu Arg Asp Ala Ser Ile Phe Gly Lys195 200 205Glu Trp Gly Leu Ser Ser Ser Glu Ile Ser Thr Phe Tyr Asn Arg Gln210 215 220Val Glu Arg Ala Gly Asp Tyr Ser Asp His Cys Val Lys Trp Tyr Ser225 230 235 240Thr Gly Leu Asn Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Val Arg245 250 255Tyr Asn Gln Phe Arg Arg Asp Met Thr Leu Met Val Leu Asp Leu Val260 265 270Ala Leu Phe Pro Ser Tyr Asp Thr Gln Met Tyr Pro Ile Lys Thr Thr275 280 285Ala Gln Leu Thr Arg Glu Val Tyr Thr Asp Ala Ile Gly Thr Val His290 295 300Pro His Pro Ser Phe Thr Ser Thr Thr Trp Tyr Asn Asn Asn Ala Pro305 310 315 320Ser Phe Ser Ala Ile Glu Ala Ala Val Val Arg Asn Pro His Leu Leu325 330 335Asp Phe Leu Glu Gln Val Thr Ile Tyr Ser Leu Leu Ser Arg Trp Ser340 345 350Asn Thr Gln Tyr Met Asn Met Trp Gly Gly His Lys Leu Glu Phe Arg355 360 365Thr Ile Gly Gly Thr Leu Asn Ile Ser Thr Gln Gly Ser Thr Asn Thr370 375 380Ser Ile Asn Pro Val Thr Leu Pro Phe Thr Ser Arg Asp Val Tyr Arg385 390 395 400Thr Glu Ser Leu Ala Gly Leu Asn Leu Phe Leu Thr Gln Pro Val Asn405 410 415Gly Val Pro Arg Val Asp Phe His Trp Lys Phe Val Thr His Pro Ile420 425 430Ala Ser Asp Asn Phe Tyr Tyr Pro Gly Tyr Ala Gly Ile Gly Thr Gln435 440 445Leu Gln Asp Ser Glu Asn Glu Leu Pro Pro Glu Ala Thr Gly Gln Pro450 455 460Asn Tyr Glu Ser Tyr Ser His Arg Leu Ser His Ile Gly Leu Ile Ser465 470 475 480Ala Ser His Val Lys Ala Leu Val Tyr Ser Trp Thr His Arg Ser Ala485 490 495Asp Arg Thr Asn Thr Ile Glu Pro Asn Ser Ile Thr Gln Ile Pro Leu500 505 510Val Lys Ala Phe Asn Leu Ser Ser Gly Ala Ala Val Val Arg Gly Pro515 520 525Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Asn Thr Gly Thr Phe530 535 540Gly Asp Ile Arg Val Asn Ile Asn Pro Pro Phe Ala Gln Arg Tyr Arg545 550 555 560Val Arg Ile Arg Tyr Ala Ser Thr Thr Asp Leu Gln Phe His Thr Ser565 570 575Ile Asn Gly Lys Ala Ile Asn Gln Gly Asn Phe Ser Ala Thr Met Asn580 585 590Arg Gly Glu Asp Leu Asp Tyr Lys Thr Phe Arg Thr Val Gly Phe Thr595 600 605Thr Pro Phe Ser Phe Leu Asp Val Gln Ser Thr Phe Thr Ile Gly Ala610 615 620Trp Asn Phe Ser Ser Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu Phe625 630 635 640Val Pro Val Glu Val Thr Tyr Glu Ala Glu Tyr Asp Phe Glu Lys Ala645 650 655Gln Glu Lys Val Thr Ala Leu Phe Thr Ser Thr Asn Pro Arg Gly Leu660 665 670Lys Thr Asp Val Lys Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val675 680 685Glu Ser Leu Ser Asp Glu Phe Tyr Leu Asp Glu Lys Arg Glu Leu Phe690 695 700Glu Ile Val Lys Tyr Ala Lys Gln Leu His Ile Glu Arg Asn Met
705 710 715<210>34<211>633<212>PRT<213>Bacillus thuringiensi<400>34Met Asn Asn Val Leu Asn Ser Gly Arg Thr Thr Ile Cys Asp Ala Tyr1 5 10 15Asn Val Val Ala His Asp Pro Phe Ser Phe G1lu His Lys Ser Leu Asp20 25 30Thr Ile Gln Lys Glu Trp Met Glu Trp Lys Arg Thr Asp His Ser Leu35 40 45Tyr Val Ala Pro Val Val Gly Thr Val Ser Ser Phe Leu Leu Lys Lys50 55 60Val Gly Ser Leu Ile Gly Lys Arg Ile Leu Ser Glu Leu Trp Gly Ile65 70 75 80Ile Phe Pro Ser Gly Ser Thr Asn Leu Met Gln Asp Ile Leu Arg Glu85 90 95Thr Glu Gln Phe Leu Asn Gln Arg Leu Asn Thr Asp Thr Leu Ala Arg100 105 110Val Asn Ala Glu Leu Ile Gly Leu Gln Ala Asn Ile Arg Glu Phe Asn115 120 125Gln Gln Val Asp Asn Phe Leu Asn Pro Thr Gln Asn Pro Val Pro Leu130 135 140Ser Ile Thr Ser Ser Val Asn Thr Met Gln Gln Leu Phe Leu Asn Arg145 150 155 160Leu Pro Gln Phe Gln Ile Gln Gly Tyr Gln Leu Leu Leu Leu Pro Leu165 170 175Phe Ala Gln Ala Ala Asn Met His Leu Ser Phe Ile Arg Asp Val Ile180 185 190Leu Asn Ala Asp Glu Trp Gly Ile Ser Ala Ala Thr Leu Arg Thr Tyr195 200 205Arg Asp Tyr Leu Arg Asn Tyr Thr Arg Asp Tyr Ser Asn Tyr Cys Ile210 215 220Asn Thr Tyr Gln Thr Ala Phe Arg Gly Leu Asn Thr Arg Leu His Asp225 230 235 240Met Leu Glu Phe Arg Thr Tyr Met Phe Leu Asn Val Phe Glu Tyr Val245 250 255Ser Ile Trp Ser Leu Phe Lys Tyr Gln Ser Leu Met Val Ser Ser Gly260 265 270Ala Asn Leu Tyr Ala Ser Gly Ser Gly Pro Gln Gln Thr Gln Ser Phe275 280 285Thr Ala Gln Asn Trp Pro Phe Leu Tyr Ser Leu Phe Gln Val Asn Ser290 295 300Asn Tyr Ile Leu Ser Gly Ile Ser Gly Thr Arg Leu Ser Ile Thr Phe305 310 315 320Pro Asn Ile Gly Gly Leu Pro Gly Ser Thr Thr Thr His Ser Leu Asn325 330 335Ser Ala Arg Val Asn Tyr Ser Gly Gly Val Ser Ser Gly Leu Ile Gly340 345 350Ala Thr Asn Leu Asn His Asn Phe Asn Cys Ser Thr Val Leu Pro Pro355 360 365Leu Ser Thr Pro Phe Val Arg Ser Trp Leu Asp Ser Gly Thr Asp Arg370 375 380Glu Gly Val Ala Thr Ser Thr Asn Trp Gln Thr Glu Ser Phe Gln Thr385 390 395 400Thr Leu Ser Leu Arg Cys Gly Ala Phe Ser Ala Arg Gly Asn Ser Asn405 410 415Tyr Phe Pro Asp Tyr Phe Ile Arg Asn Ile Ser Gly Val Pro Leu Val420 425 430Ile Arg Asn Glu Asp Leu Thr Arg Pro Leu His Tyr Asn Gln Ile Arg435 440 445Asn Ile Glu Ser Pro Ser Gly Thr Pro Gly Gly Ala Arg Ala Tyr Leu450 455 460Val Ser Val His Asn Arg Lys Asn Asn Ile Tyr Ala Ala Asn Glu Asn
465 470 475 480Gly Thr Met Ile His Leu Ala Pro Glu Asp Tyr Thr Gly Phe Thr Ile485 490 495Ser Pro Ile His Ala Thr Gln Val Asn Asn Gln Thr Arg Thr Phe Ile500 505 510Ser Glu Lys Phe Gly Asn Gln Gly Asp Ser Leu Arg Phe Glu Gln Ser515 520 525Asn Thr Thr Ala Arg Tyr Thr Leu Arg Gly Asn Gly Asn Ser Tyr Asn530 535 540Leu Tyr Leu Arg Val Ser Ser Ile Gly Asn Ser Thr Ile Arg Val Thr545 550 555 560Ile Asn Gly Arg Val Tyr Thr Val Ser Asn Val Asn Thr Thr Thr Asn565 570 575Asn Asp Gly Val Asn Asp Asn Gly Ala Arg Phe Ser Asp Ile Asn Ile580 585 590Gly Asn Ile Val Ala Ser Asp Asn Thr Asn Val Thr Leu Asp Ile Asn595 600 605Val Thr Leu Asn Ser Gly Thr Pro Phe Asp Leu Met Asn Ile Met Phe610 615 620Val Pro Thr Asn Leu Pro Pro Leu Tyr625 630<210>35<211>652<212>PRT<213>Bacillus thuringiensi<400>35Met Ile Arg Lys Gly Gly Arg Lys Met Asn Pro Asn Asn Arg Ser Glu1 5 10 15His Asp Thr Ile Lys Thr Thr Glu Asn Asn Glu Val Pro Thr Asn His20 25 30Val Gln Tyr Pro Leu Ala Glu Thr Pro Asn Pro Thr Leu Glu Asp Leu35 40 45Asn Tyr Lys Glu Phe Leu Arg Met Thr Ala Asp Asn Asn Thr Glu Ala50 55 60Leu Asp Ser Ser Thr Thr Lys Asp Val Ile Gln Lys Gly Ile Ser Val65 70 75 80Val Gly Asp Leu Leu Gly Val Val Gly Phe Pro Phe Gly Gly Ala Leu85 90 95Val Ser Phe Tyr Thr Asn Phe Leu Asn Thr Ile Trp Pro Ser Glu Asp100 105 110Pro Trp Lys Ala Phe Met Glu Gln Val Glu Ala Leu Met Asp Gln Lys115 120 125Ile Ala Asp Tyr Ala Lys Asn Lys Ala Leu Ala Glu Leu Gln Gly Leu130 135 140Gln Asn Asn Val Glu Asp Tyr Val Ser Ala Leu Ser Ser Trp Gln Lys145 150 155 160Asn Pro Val Ser Ser Arg Asn Pro His Ser Gln Gly Arg Ile Arg Glu165 170 175Leu Phe Ser Gln Ala Glu Ser His Phe Arg Asn Ser Met Pro Ser Phe180 185 190Ala Ile Ser Gly Tyr Glu Val Leu Phe Leu Thr Thr Tyr Ala Gln Ala195 200 205Ala Asn Thr His Leu Phe Leu Leu Lys Asp Ala Gln Ile Tyr Gly Glu210 215 220Glu Trp Gly Tyr Glu Lys Glu Asp Ile Ala Glu Phe Tyr Lys Arg Gln225 230 235 240Leu Lys Leu Thr Gln Glu Tyr Thr Asp His Cys Val Lys Trp Tyr Asn245 250 255Val Gly Leu Asp Lys Leu Arg Gly Ser Ser Tyr Glu Ser Trp Val Asn260 265 270Phe Asn Arg Tyr Arg Arg Glu Met Thr Leu Thr Val Leu Asp Leu Ile275 280 285Ala Leu Phe Pro Leu Tyr Asp Val Arg Leu Tyr Pro Lys Glu Val Lys290 295 300Thr Glu Leu Thr Arg Asp Val Leu Thr Asp Pro Ile Val Gly Val Asn
305 310 315 320Asn Leu Arg Gly Tyr Gly Thr Thr Phe Ser Asn Ile Glu Asn Tyr Ile325 330 335Arg Lys Pro His Leu Phe Asp Tyr Leu His Arg Ile Gln Phe His Thr340 345 350Arg Phe Gln Pro Gly Tyr Tyr Gly Asn Asp Ser Phe Asn Tyr Trp Ser355 360 365Gly Asn Tyr Val Ser Thr Arg Pro Ser Ile Gly Ser Asn Asp Ile Ile370 375 380TLr Ser Pro Phe Tyr Gly Asn Lys Ser Ser Glu Pro Val Gln Asn Leu385 390 395 400Glu Phe Asn Gly Glu Lys Val Tyr Arg Ala Val Ala Asn Thr Asn Leu405 410 415Ala Val Trp Pro Ser Ala Val Tyr Ser Gly Val Thr Lys Val Glu Phe420 425 430Ser Gln Tyr Asn Asp Gln Thr Asp Glu Ala Ser Thr Gln Thr Tyr Asp435 440 445Ser Lys Arg Asn Val Gly Ala Val Ser Trp Asp Ser Ile Asp Gln Leu450 455 460Pro Pro Glu Thr Thr Asp Glu Pro Leu Glu Lys Gly Tyr Ser His Gln465 470 475 480Leu Asn Tyr Val Met Cys Phe Leu Met Gln Gly Ser Arg Gly Thr Ile485 490 495Pro Val Leu Thr Trp Thr His Lys Ser Val Asp Phe Phe Asn Met Ile500 505 510Asp Ser Lys Lys Ile Thr Gln Leu Pro Leu Val Lys Ala Tyr Lys Leu515 520 525Gln Ser Gly Ala Ser Val Val Ala Gly Pro Arg Phe Thr Gly Gly Asp530 535 540Ile Ile Gln Cys Thr Glu Asn Gly Ser Ala Ala Thr Ile Tyr Val Thr545 550 555 560Pro Asp Val Ser Tyr Ser Gln Lys Tyr Arg Ala Arg Ile His Tyr Ala565 570 575Ser Thr Ser Gln Ile Thr Phe Thr Leu Ser Leu Asp Gly Ala Pro Phe580 585 590Asn Gln Tyr Tyr Phe Asp Lys Thr Ile Asn Lys Gly Asp Thr Leu Thr595 600 605Tyr Asn Ser Phe Asn Leu Ala Ser Phe Ser Thr Pro Phe Glu Leu Ser610 615 620Gly Asn Asn Leu Gln Ile Gly Val Thr Gly Leu Ser Ala Gly Asp Lys625 630 635 640Val Tyr Ile Asp Lys Ile Glu Phe Ile Pro Val Asn645 650<210>36<211>659<212>PRT<213>Bacillus thuringiensi<400>36Met Ile Arg Met Gly Gly Arg Lys Met Asn Pro Asn Asn Arg Ser Glu1 5 10 15Tyr Asp Thr Ile Lys Val Thr Pro Asn Ser Glu Leu Pro Thr Asn His20 25 30Asn Gln Tyr Pro Leu Ala Asp Asn Pro Asn Ser Thr Leu Glu Glu Leu35 40 45Asn Tyr Lys Glu Phe Leu Arg Met Thr Ala Asp Asn Ser Thr Glu Val50 55 60Leu Asp Ser Ser Thr Val Lys Asp Ala Val Gly Thr Gly Ile Ser Val65 70 75 80Val Gly Gln Ile Leu Gly Val Val Gly Val Pro Phe Ala Gly Ala Leu85 90 95Thr Ser Phe Tyr Gln Ser Phe Leu Asn Ala Ile Trp Pro Ser Asp Ala100 105 110Asp Pro Trp Lys Ala Phe Met Ala Gln Val Glu Val Leu Ile Asp Lys115 120 125Lys Ile Glu Glu Tyr Ala Lys Ser Lys Ala Leu Ala Glu Leu Gln Gly
130 135 140Leu Gln Asn Asn Phe Glu Asp Tyr Val Asn Ala Leu Asp Ser Trp Lys145 150 155 160Lys Ala Pro Val Asn Leu Arg Ser Arg Arg Ser Gln Asp Arg Ile Arg165 170 175Glu Leu Phe Ser Gln Ala Glu Ser His Phe Arg Asn Ser Met Pro Ser180 185 190Phe Ala Val Ser Lys Phe Glu Val Leu Phe Leu Pro Thr Tyr Ala Gln195 200 205Ala Ala Asn Thr His Leu Leu Leu Leu Lys Asp Ala Gln Val Phe Gly210 215 220Glu Glu Trp Gly Tyr Ser Ser Glu Asp Ile Ala Glu Phe Tyr Gln Arg225 230 235 240Gln Leu Lys Leu Thr Gln Gln Tyr Thr Asp His Cys Val Asn Trp Tyr245 250 255Asn Val Gly Leu Asn Ser Leu Arg Gly Ser Thr Tyr Asp Ala Trp Val260 265 270Lys Phe Asn Arg Phe Arg Arg Glu Met Thr Leu Thr Val Leu Asp Leu275 280 285Ile Val Leu Phe Pro Phe Tyr Asp Val Arg Leu Tyr Ser Lys Gly Val290 295 300Lys Thr Glu Leu Thr Arg Asp Ile Phe Thr Asp Pro Ile Phe Thr Leu305 310 315 320Asn Ala Leu Gln Glu Tyr Gly Pro Thr Phe Ser Ser Ile Glu Asn Ser325 330 335Ile Arg Lys Pro His Leu Phe Asp Tyr Leu Arg Gly Ile Glu Phe His340 345 350Thr Arg Leu Arg Pro Gly Tyr Ser Gly Lys Asp Ser Phe Asn Tyr Trp355 360 365Ser Gly Asn Tyr Val Glu Thr Arg Pro Ser Ile Gly Ser Asn Asp Thr370 375 380Ile Thr Ser Pro Phe Tyr Gly Asp Lys Ser Ile Glu Pro Ile Gln Lys385 390 395 400Leu Ser Phe Asp Gly Gln Lys Val Tyr Arg Thr Ile Ala Asn Thr Asp405 410 415Ile Ala Ala Phe Pro Asp Gly Lys Ile Tyr Phe Gly Val Thr Lys Val420 425 430Asp Phe Ser Gln Tyr Asp Asp Gln Lys Asn Glu Thr Ser Thr Gln Thr435 440 445Tyr Asp Ser Lys Arg Tyr Asn Gly Tyr Leu Gly Ala Gln Asp Ser Ile450 455 460Asp Gln Leu Pro Pro Glu Thr Thr Asp Glu Pro Leu Glu Lys Ala Tyr465 470 475 480Ser His Gln Leu Asn Tyr Ala Glu Cys Phe Leu Met Gln Asp Arg Arg485 490 495Gly Thr Ile Pro Phe Phe Thr Trp Thr His Arg Ser Val Asp Phe Phe500 505 510Asn Thr Ile Asp Ala Glu Lys Ile Thr Gln Leu Pro Val Val Lys Ala515 520 525Tyr Ala Leu Ser Ser Gly Ala Ser Ile Ile Glu Gly Pro Gly Phe Thr530 535 540Gly Gly Asn Leu Leu Phe Leu Lys Glu Ser Ser Asn Ser Ile Ala Lys545 550 555 560Phe Lys Val Thr Leu Asn Ser Ala Ala Leu Leu Gln Arg Tyr Arg Val565 570 575Arg Ile Arg Tyr Ala Ser Thr Thr Asn Leu Arg Leu Phe Val Gln Asn580 585 590Ser Asn Asn Asp Phe Leu Val Ile Tyr Ile Asn Lys Thr Met Asn Ile595 600 605Asp Gly Asp Leu Thr Tyr Gln Thr Phe Asp Phe Ala Thr Ser Asn Ser610 615 620Asn Met Gly Phe Ser Gly Asp Thr Asn Asp Phe Ile Ile Gly Ala Glu625 630 635 640Ser Phe Val Ser Asn Glu Lys Ile Tyr Ile Asp Lys Ile Glu Phe Ile645 650 655Pro Val Gln
<210>37<211>652<212>PRT<213>Bacillus thuringiensi<400>37Met Asn Pro Asn Asn Arg Ser Glu His Asp Thr Ile Lys Val Thr Pro1 5 10 15Asn Ser Glu Leu Gln Thr Asn His Asn Gln Tyr Pro Leu Ala Asp Asn20 25 30Pro Asn Ser Thr Leu Glu Glu Leu Asn Tyr Lys Glu Phe Leu Arg Met35 40 45Thr Glu Asp Ser Ser Thr Glu Val Leu Asp Asn Ser Thr Val Lys Asp50 55 60Ala Val Gly Thr Gly Ile Ser Val Val Gly Gln Ile Leu Gly Val Val65 70 75 80Gly Val Pro Phe Ala Gly Ala Leu Thr Ser Phe Tyr Gln Ser Phe Leu85 90 95Asn Thr Ile Trp Pro Ser Asp Ala Asp Pro Trp Lys Ala Phe Met Ala100 105 110Gln Val Glu Val Leu Ile Asp Lys Lys Ile Glu Glu Tyr Ala Lys Ser115 120 125Lys Ala Leu Ala Glu Leu Gln Gly Leu Gln Asn Asn Phe Glu Asp Tyr130 135 140Val Asn Ala Leu Asn Ser Trp Lys Lys Thr Pro Leu Ser Leu Arg Ser145 150 155 160Lys Arg Ser Gln Asp Arg Ile Arg Glu Leu Phe Ser Gln Ala Glu Ser165 170 175His Phe Arg Asn Ser Met Pro Ser Phe Ala Val Ser Lys Phe Glu Val180 185 190Leu Phe Leu Pro Thr Tyr Ala Gln Ala Ala Asn Thr His Leu Leu Leu195 200 205Leu Lys Asp Ala Gln Val Phe Gly Glu Glu Trp Gly Tyr Ser Ser Glu210 215 220Asp Val Ala Glu Phe Tyr His Arg Gln Leu Lys Leu Thr Gln Gln Tyr225 230 235 240Thr Asp His Cys Val Asn Trp Tyr Asn Val Gly Leu Asn Gly Leu Arg245 250 255Gly Ser Thr Tyr Asp Ala Trp Val Lys Phe Asn Arg Phe Arg Arg Glu260 265 270Met Thr Leu Thr Val Leu Asp Leu Ile Val Leu Phe Pro Phe Tyr Asp275 280 285Ile Arg Leu Tyr Ser Lys Gly Val Lys Thr Glu Leu Thr Arg Asp Ile290 295 300Phe Thr Asp Pro Ile Phe Ser Leu Asn Thr Leu Gln Glu Tyr Gly Pro305 310 315 320Thr Phe Leu Ser Ile Glu Asn Ser Ile Arg Lys Pro His Leu Phe Asp325 330 335Tyr Leu Gln Gly Ile Glu Phe His Thr Arg Leu Gln Pro Gly Tyr Phe340 345 350Gly Lys Asp Ser Phe Asn Tyr Trp Ser Gly Asn Tyr Val Glu Thr Arg355 360 365Pro Ser Ile Gly Ser Ser Lys Thr Ile Thr Ser Pro Phe Tyr Gly Asp370 375 380Lys Ser Thr Glu Pro Val Gln Lys Leu Ser Phe Asp Gly Gln Lys Val385 390 395 400Tyr Arg Thr Ile Ala Asn Thr Asp Val Ala Ala Trp Pro Asn Gly Lys405 410 415Val Tyr Leu Gly Val Thr Lys Val Asp Phe Ser Gln Tyr Asp Asp Gln420 425 430Lys Asn Glu Thr Ser Thr Gln Thr Tyr Asp Ser Lys Arg Asn Asn Gly435 440 445His Val Ser Ala Gln Asp Ser Ile Asp Gln Leu Pro Pro Glu Thr Thr450 455 460Asp Glu Pro Leu Glu Lys Ala Tyr Ser His Gln Leu Asn Tyr Ala Glu465 470 475 480Cys Phe Leu Met Gln Asp Arg Arg Gly Thr Ile Pro Phe Phe Thr Trp
485 490 495Thr His Arg Ser Val Asp Phe Phe Asn Thr Ile Asp Ala Glu Lys Ile500 505 510Thr Gln Leu Pro Val Val Lys Ala Tyr Ala Leu Ser Ser Gly Ala Ser515 520 525Ile Ile Glu Gly Pro Gly Phe Thr Gly Gly Asn Leu Leu Phe Leu Lys530 535 540Glu Ser Ser Asn Ser Ile Ala Lys Phe Lys Val Thr Leu Asn Ser Ala545 550 555 560Ala Leu Leu Gln Arg Tyr Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr565 570 575Asn Leu Arg Leu Phe Val Gln Asn Ser Asn Asn Asp Phe Leu Val Ile580 585 590Tyr Ile Asn Lys Thr Met Asn Lys Asp Asp Asp Leu Thr Tyr Gln Thr595 600 605Phe Asp Leu Ala Thr Thr Asn Ser Asn Met Gly Phe Ser Gly Asp Lys610 615 620Asn Glu Leu Ile Ile Gly Ala Glu Ser Phe Val Ser Asn Glu Lys Ile625 630 635 640Tyr Ile Asp Lys Ile Glu Phe Ile Pro Val Gln Leu645 650<210>38<211>1180<212>PRT<213>Bacillus thuringiensi<400>38Met Asn Pro Tyr Gln Asn Lys Asn Glu Tyr Glu Thr Leu Asn Ala Ser1 5 10 15Gln Lys Lys Leu Asn Ile Ser Asn Asn Tyr Thr Arg Tyr Pro Ile Glu20 25 30Asn Ser Pro Lys Gln Leu Leu Gln Ser Thr Asn Tyr Lys Asp Trp Leu35 40 45Asn Met Cys Gln Gln Asn Gln Gln Tyr Gly Gly Asp Phe Glu Thr Phe50 55 60Ile Asp Ser Gly Glu Leu Ser Ala Tyr Thr Ile Val Val Gly Thr Val65 70 75 80Leu Thr Gly Phe Gly Phe Thr Thr Pro Leu Gly Leu Ala Leu Ile Gly85 90 95Phe Gly Thr Leu Ile Pro Val Leu Phe Pro Ala Gln Asp Gln Ser Asn100 105 110Thr Trp Ser Asp Phe Ile Thr Gln Thr Lys Asn Ile Ile Lys Lys Glu115 120 125Ile Ala Ser Thr Tyr Ile Ser Asn Ala Asn Lys Ile Leu Asn Arg Ser130 135 140Phe Asn Val Ile Ser Thr Tyr His Asn His Leu Lys Thr Trp Glu Asn145 150 155 160Asn Pro Asn Pro Gln Asn Thr Gln Asp Val Arg Thr Gln Ile Gln Leu165 170 175Val His Tyr His Phe Gln Asn Val Ile Pro Glu Leu Val Asn Ser Cys180 185 190Pro Pro Asn Pro Ser Asp Cys Asp Tyr Tyr Asn Ile Leu Val Leu Ser195 200 205Ser Tyr Ala Gln Ala Ala Asn Leu His Leu Thr Val Leu Asn Gln Ala210 215 220Val Lys Phe Glu Ala Tyr Leu Lys Asn Asn Arg Gln Phe Asp Tyr Leu225 230 235 240Glu Pro Leu Pro Thr Ala Ile Asp Tyr Tyr Pro Val Leu Thr Lys Ala245 250 255Ile Glu Asp Tyr Thr Asn Tyr Cys Val Thr Thr Tyr Lys Lys Gly Leu260 265 270Asn Leu Ile Lys Thr Thr Pro Asp Ser Asn Leu Asp Gly Asn Ile Asn275 280 285Trp Asn Thr Tyr Asn Thr Tyr Arg Thr Lys Met Thr Thr Ala Val Leu290 295 300Asp Leu Val Ala Leu Phe Pro Asn Tyr Asp Val Gly Lys Tyr Pro Ile
305 310 315 320Gly Val Gln Ser Glu Leu Thr Arg Glu Ile Tyr Gln Val Leu Asn Phe325 330 335Glu Glu Ser Pro Tyr Lys Tyr Tyr Asp Phe Gln Tyr Gln Glu Asp Ser340 345 350Leu Thr Arg Arg Pro His Leu Phe Thr Trp Leu Asp Ser Leu Asn Phe355 360 365Tyr Glu Lys Ala Gln Thr Thr Pro Asn Asn Phe Phe Thr Ser His Tyr370 375 380Asn Met Phe His Tyr Thr Leu Asp Asn Ile Ser Gln Lys Ser Ser Val385 390 395 400Phe Gly Asn His Asn Val Thr Asp Lys Leu Lys Ser Leu Gly Leu Ala405 410 415Thr Asn Ile Tyr Ile Phe Leu Leu Asn Val Ile Ser Leu Asp Asn Lys420 425 430Tyr Leu Asn Asp Tyr Asn Asn Ile Ser Lys Met Asp Phe Phe Ile Thr435 440 445Asn Gly Thr Arg Leu Leu Glu Lys Glu Leu Thr Ala Gly Ser Gly Gln450 455 460Ile Thr Tyr Asp Val Asn Lys Asn Ile Phe Gly Leu Pro Ile Leu Lys465 470 475 480Arg Arg Glu Asn Gln Gly Asn Pro Thr Leu Phe Pro Thr Tyr Asp Asn485 490 495Tyr Ser His Ile Leu Ser Phe Ile Lys Ser Leu Ser Ile Pro Ala Thr500 505 510Tyr Lys Thr Gln Val Tyr Thr Phe Ala Trp Thr His Ser Ser Val Asp515 520 525Pro Lys Asn Thr Ile Tyr Thr His Leu Thr Thr Gln Ile Pro Ala Val530 535 540Lys Ala Asn Ser Leu Gly Thr Ala Ser Lys Val Val Gln Gly Pro Gly545 550 555 560His Thr Gly Gly Asp Leu Ile Asp Phe Lys Asp His Phe Lys Ile Thr565 570 575Cys Gln His Ser Asn Phe Gln Gln Ser Tyr Phe Ile Arg Ile Arg Tyr580 585 590Ala Ser Asn Gly Ser Ala Asn Thr Arg Ala Val Ile Asn Leu Ser Ile595 600 605Pro Gly Val Ala Glu Leu Gly Met Ala Leu Asn Pro Thr Phe Ser Gly610 615 620Thr Asp Tyr Thr Asn Leu Lys Tyr Lys Asp Phe Gln Tyr Leu Glu Phe625 630 635 640Ser Asn Glu Val Lys Phe Ala Pro Asn Gln Asn Ile Ser Leu Val Phe645 650 655Asn Arg Ser Asp Val Tyr Thr Asn Thr Thr Val Leu Ile Asp Lys Ile660 665 670Glu Phe Leu Pro Ile Thr Arg Ser Ile Arg Glu Asp Arg Glu Lys Gln675 680 685Lys Leu Glu Thr Val Gln Gln Ile Ile Asn Thr Phe Tyr Ala Asn Pro690 695 700Ile Lys Asn Thr Leu Gln Ser Glu Leu Thr Asp Tyr Asp Ile Asp Gln705 710 715 720Ala Ala Asn Leu Val Glu Cys Ile Ser Glu Glu Leu Tyr Pro Lys Glu725 730 735Lys Met Leu Leu Leu Asp Glu Val Lys Asn Ala Lys Gln Leu Ser Gln740 745 750Ser Arg Asn Val Leu Gln Asn Gly Asp Phe Glu Ser Ala Thr Leu Gly755 760 765Trp Thr Thr Ser Asp Asn Ile Thr Ile Gln Glu Asp Asp Pro Ile Phe770 775 780Lys Gly His Tyr Leu His Met Ser Gly Ala Arg Asp Ile Asp Gly Thr785 790 795 800Ile Phe Pro Thr Tyr Ile Phe Gln Lys Ile Asp Glu Ser Lys Leu Lys805 810 815Pro Tyr Thr Arg Tyr Leu Val Arg Gly Phe Val Gly Ser Ser Lys Asp820 825 830Val Glu Leu Val Val Ser Arg Tyr Gly Glu Glu Ile Asp Ala Ile Met835 840 845Asn Val Pro Ala Asp Leu Asn Tyr Leu Tyr Pro Ser Thr Phe Asp Cys
850 855 860Glu Gly Ser Asn Arg Cys Glu Thr Ser Ala Val Pro Ala Asn Ile Gly865 870 875 880Asn Thr Ser Asp Met Leu Tyr Ser Cys Gln Tyr Asp Thr Gly Lys Lys885 890 895His Val Val Cys Gln Asp Ser His Gln Phe Ser Phe Thr Ile Asp Thr900 905 910Gly Ala Leu Asp Thr Asn Glu Asn Ile Gly Val Trp Val Met Phe Lys915 920 925Ile Ser Ser Pro Asp Gly Tyr Ala Ser Leu Asp Asn Leu Glu Val Ile930 935 940Glu Glu Gly Pro Ile Asp Gly Glu Ala Leu Ser Arg Val Lys His Met945 950 955 960Glu Lys Lys Trp Asn Asp Gln Met Glu Ala Lys Arg Ser Glu Thr Gln965 970 975Gln Ala Tyr Asp Val Ala Lys Gln Ala Ile Asp Ala Leu Phe Thr Asn980 985 990Val Gln Asp Glu Ala Leu Gln Phe Asp Thr Thr Leu Ala Gln Ile Gln99510001005Tyr Ala Glu Tyr Leu Val Gln Ser Ile Pro Tyr Val Tyr Asn Asp Trp101010151020Leu Ser Asp Val Pro Gly Met Asn Tyr Asp Ile Tyr Val Glu Leu Asp102510301035 1040Ala Arg Val Ala Gln Ala Arg Tyr Leu Tyr Asp Thr Arg Asn Ile Ile104510501055Lys Asn Gly Asp Phe Thr Gln Gly Val Met Gly Trp His Val Thr Gly106010651070Asn Ala Asp Val Gln Gln Ile Asp Gly Val Ser Val Leu Val Leu Ser107510801085Asn Trp Ser Ala Gly Val Ser Gln Asn Val His Leu Gln His Asn His109010951100Gly Tyr Val Leu Arg Val Ile Ala Lys Lys Glu Gly Pro Gly Asn Gly110511101115 1120Tyr Val Thr Leu Met Asp Cys Glu Glu Asn Gln Glu Lys Leu Thr Phe112511301135Thr Ser Cys Glu Glu Gly Tyr Ile Thr Lys Thr Val Asp Val Phe Pro114011451150Asp Thr Asp Arg Val Arg Ile Glu Ile Gly Glu Thr Glu Gly Ser Phe115511601165Tyr Ile Glu Ser Ile Glu Leu Ile Cys Met Asn Glu117011751180<210>39<211>1136<212>PRT<213>Bacillus thuringiensi<400>39Met Asn Ser Gly Tyr Pro Leu Ala Asn Asp Leu Gln Gly Ser Met Lys1 5 10 15Asn Thr Asn Tyr Lys Asp Trp Leu Ala Met Cys Glu Asn Asn Gln Gln20 25 30Tyr Gly Val Asn Pro Ala Ala Ile Asn Ser Ser Ser Val Ser Thr Ala35 40 45Leu Lys Val Ala Gly Ala Ile Leu Lys Phe Val Asn Pro Pro Ala Gly50 55 60Thr Val Leu Thr Val Leu Ser Ala Val Leu Pro Ile Leu Trp Pro Thr65 70 75 80Asn Thr Pro Thr Pro Glu Arg Val Trp Asn Asp Phe Met Thr Asn Thr85 90 95Gly Asn Leu Ile Asp Gln Thr Val Thr Ala Tyr Val Arg Thr Asp Ala100 105 110Asn Ala Lys Met Thr Val Val Lys Asp Tyr Leu Asp Gln Tyr Thr Thr115 120 125Lys Phe Asn Thr Trp Lys Arg Glu Pro Asn Asn Gln Ser Tyr Arg Thr130 135 140Ala Val Ile Thr Gln Phe Asn Leu Thr Ser Ala Lys Leu Arg Glu Thr
145 150 155 160Ala Val Tyr Phe Ser Asn Leu Val Gly Tyr Glu Leu Leu Leu Leu Pro165 170 175Ile Tyr Ala Gln Val Ala Asn Phe Asn Leu Leu Leu Ile Arg Asp Gly180 185 190Leu Ile Asn Ala Gln Glu Trp Ser Leu Ala Arg Ser Ala Gly Asp Gln195 200 205Leu Tyr Asn Thr Met Val Gln Tyr Thr Lys Glu Tyr Ile Ala His Ser210 215 220Ile Thr Trp Tyr Asn Lys Gly Leu Asp Val Leu Arg Asn Lys Ser Asn225 230 235 240Gly Gln Trp Ile Thr Phe Asn Asp Tyr Lys Arg Glu Met Thr Ile Gln245 250 255Val Leu Asp Ile Leu Ala Leu Phe Ala Ser Tyr Asp Pro Arg Arg Tyr260 265 270Pro Ala Asp Lys Ile Asp Asn Thr Lys Leu Ser Lys Thr Glu Phe Thr275 280 285Arg Glu Ile Tyr Thr Ala Leu Val Glu Ser Pro Ser Ser Lys Ser Ile290 295 300Ala Ala Leu Glu Ala Ala Leu Thr Arg Asp Val His Leu Phe Thr Trp305 310 315 320Leu Lys Arg Val Asp Phe Trp Thr Asn Thr Ile Tyr Gln Asp Leu Arg325 330 335Phe Leu Ser Ala Asn Lys Ile Gly Phe Ser Tyr Thr Asn Ser Ser Ala340 345 350Met Gln Glu Ser Gly Ile Tyr Gly Ser Ser Gly Phe Gly Ser Asn Leu355 360 365Thr His Gln Ile Gln Leu Asn Ser Asn Val Tyr Lys Thr Ser Ile Thr370 375 380Asp Thr Ser Ser Pro Ser Asn Arg Val Thr Lys Met Asp Phe Tyr Lys385 390 395 400Ile Asp Gly Thr Leu Ala Ser Tyr Asn Ser Asn Ile Thr Pro Thr Pro405 410 415Glu Gly Leu Arg Thr Thr phe Phe Gly Phe Ser Thr Asn Glu Asn Thr420 425 430Pro Asn Gln Pro Thr Val Asn Asp Tyr Thr His Ile Leu Ser Tyr Ile435 440 445Lys Thr Asp Val Ile Asp Tyr Asn Ser Asn Arg Val Ser Phe Ala Trp450 455 460Thr His Lys Ile Val Asp Pro Asn Asn Gln Ile Tyr Thr Asp Ala Ile465 470 475 480Thr Gln Val Pro Ala Val Lys Ser Asn Phe Leu Asn Ala Thr Ala Lys485 490 495Val Ile Lys Gly Pro Gly His Thr Gly Gly Asp Leu Val Ala Leu Thr500 505 510Ser Asn Gly Thr Leu Ser Gly Arg Met Glu Ile Gln Cys Lys Thr Ser515 520 525Ile Phe Asn Asp Pro Thr Arg Ser Tyr Gly Leu Arg Ile Arg Tyr Ala530 535 540Ala Asn Ser Pro Ile Val Leu Asn Val Ser Tyr Val Leu Gln Gly Val545 550 555 560Ser Arg Gly Thr Thr Ile Ser Thr Glu Ser Thr Phe Ser Arg Pro Asn565 570 575Asn Ile Ile Pro Thr Asp Leu Lys Tyr Glu Glu Phe Arg Tyr Lys Asp580 585 590Pro Phe Asp Ala Ile Val Pro Met Arg Leu Ser Ser Asn Gln Leu Ile595 600 605Thr Ile Ala Ile Gln Pro Leu Asn Met Thr Ser Asn Asn Gln Val Ile610 615 620Ile Asp Arg Ile Glu Ile Ile Pro Ile Thr Gln Ser Val Leu Asp Glu625 630 635 640Thr Glu Asn Gln Asn Leu Glu Ser Glu Arg Glu Val Val Asn Ala Leu645 650 655Phe Thr Asn Asp Ala Lys Asp Ala Leu Asn Ile Gly Thr Thr Asp Tyr660 665 670Asp Ile Asp Gln Ala Ala Asn Leu Val Glu Cys Ile Ser Glu Glu Leu675 680 685Tyr Pro Lys Glu Lys Met Leu Leu Leu Asp Glu Val Lys Asn Ala Lys
690 695 700Gln Leu Ser Gln Ser Arg Asn Val Leu Gln Asn Gly Asp Phe Glu Ser705 710 715 720Ala Thr Leu Gly Trp Thr Thr Ser Asp Asn Ile Thr Ile Gln Glu Asp725 730 735Asp Pro Ile Phe Lys Gly His Tyr Leu His Met Ser Gly Ala Arg Asp740 745 750Ile Asp Gly Thr Ile Phe Pro Thr Tyr Ile Phe Gln Lys Ile Asp Glu755 760 765Ser Lys Leu Lys Pro Tyr Thr Arg Tyr Leu Val Arg Gly Phe Val Gly770 775 780Ser Ser Lys Asp Val Glu Leu Val Val Ser Arg Tyr Gly Glu Glu Ile785 790 795 800Asp Ala Ile Met Asn Val Pro Ala Asp Leu Asn Tyr Leu Tyr Pro Ser805 810 815Thr Phe Asp Cys Glu Gly Ser Asn Arg Cys Glu Thr Ser Ala Val Pro820 825 830Ala Asn Ile Gly Asn Thr Ser Asp Met Leu Tyr Ser Cys Gln Tyr Asp835 840 845Thr Gly Lys Lys His Val Val Cys Gln Asp Ser His Gln Phe Ser Phe850 855 860Thr Ile Asp Thr Gly Ala Leu Asp Thr Asn Glu Asn Ile Gly Val Trp865 870 875 880Val Met Phe Lys Ile Ser Ser Pro Asp Gly Tyr Ala Ser Leu Asp Asn885 890 895Leu Glu Val Ile Glu Glu Gly Pro Ile Asp Gly Glu Ala Leu Ser Arg900 905 910Val Lys His Met Glu Lys Lys Trp Asn Asp Gln Met Glu Ala Lys Arg915 920 925Ser Glu Thr Gln Gln Ala Tyr Asp Val Ala Lys Gln Ala Ile Asp Ala930 935 940Leu Phe Thr Asn Val Gln Asp Glu Ala Leu Gln Phe Asp Thr Thr Leu945 950 955 960Ala Gln Ile Gln Tyr Ala Glu Tyr Leu Val Gln Ser Ile Pro Tyr Val965 970 975Tyr Asn Asp Trp Leu Ser Asp Val Pro Gly Met Asn Tyr Asp Ile Tyr980 985 990Val Glu Leu Asp Ala Arg Val Ala Gln Ala Arg Tyr Leu Tyr Asp Thr995 10001005Arg Asn Ile Ile Lys Asn Gly Asp Phe Thr Gln Gly Val Met Gly Trp101010151020His Val Thr Gly Asn Ala Asp Val Gln Gln Ile Asp Gly Val Ser Val102510301035 1040Leu Val Leu Ser Asn Trp Ser Ala Gly Val Ser Gln Asn Val His Leu104510501055Gln His Asn His Gly Tyr Val Leu Arg Val Ile Ala Lys Lys Glu Gly106010651070Pro Gly Asn Gly Tyr Val Thr Leu Met Asp Cys Glu Glu Asn Gln Glu107510801085Lys Leu Thr Phe Thr Ser Cys Glu Glu Gly Tyr Ile Thr Lys Thr Val109010951100Asp Val Phe Pro Asp Thr Asp Arg Val Arg Ile Glu Ile Gly Glu Thr110511101115 1120Glu Gly Ser Phe Tyr Ile Glu Ser Ile Glu Leu Ile Cys Met Asn Glu112511301135<210>40<211>475<212>PRT<213>Bacillus thuringiensi<400>40Met Ile Ile Asp Ser Lys Thr Thr Leu Pro Arg His Ser Leu Ile His1 5 10 15Thr Ile Lys Leu Asn Ser Asn Lys Lys Tyr Gly Pro Gly Asp Met Thr20 25 30Asn Gly Asn Gln Phe Ile Ile Ser Lys Gln Glu Trp Ala Thr Ile Gly
35 40 45Ala Tyr Ile Gln Thr Gly Leu Gly Leu Pro Val Asn Glu Gln Gln Leu50 55 60Arg Thr His Val Asn Leu Ser Gln Asp Ile Ser Ile Pro Ser Asp Phe65 70 75 80Ser Gln Leu Tyr Asp Val Tyr Cys Ser Asp Lys Thr Ser Ala Glu Trp85 90 95Trp Asn Lys Asn Leu Tyr Pro Leu Ile Ile Lys Ser Ala Asn Asp Ile100 105 110Ala Ser Tyr Gly Phe Lys Val Ala Gly Asp Pro Ser Ile Lys Lys Asp115 120 125Gly Tyr Phe Lys Lys Leu Gln Asp Glu Leu Asp Asn Ile Val Asp Asn130 135 140Asn Ser Asp Asp Asp Ala Ile Ala Lys Ala Ile Lys Asp Phe Lys Ala145 150 155 160Arg Cys Gly Ile Leu Ile Lys Glu Ala Lys Gln Tyr Glu Glu Ala Ala165 170 175Lys Asn Ile Val Thr Ser Leu Asp Gln Phe Leu His Gly Asp Gln Lys180 185 190Lys Leu Glu Gly Val Ile Asn Ile Gln Lys Arg Leu Lys Glu Val Gln195 200 205Thr Ala Leu Asn Gln Ala His Gly Glu Ser Ser Pro Ala His Lys Glu210 215 220Leu Leu Glu Lys Val Lys Asn Leu Lys Thr Thr Leu Glu Arg Thr Ile225 230 235 240Lys Ala Glu Gln Asp Leu Glu Lys Lys Val Glu Tyr Ser Phe Leu Leu245 250 255Gly Pro Leu Leu Gly Phe Val Val Tyr Glu Ile Leu Glu Asn Thr Ala260 265 270Val Gln His Ile Lys Asn Gln Ile Asp Glu Ile Lys Lys Gln Leu Asp275 280 285Ser Ala Gln His Asp Leu Asp Arg Asp Val Lys Ile Ile Gly Met Leu290 295 300Asn Ser Ile Asn Thr Asp Ile Asp Asn Leu Tyr Ser Gln Gly Gln Glu305 310 315 320Ala Ile Lys Val Phe Gln Lys Leu Gln Gly Ile Trp Ala Thr Ile Gly325 330 335Ala Gln Ile Glu Asn Leu Arg Thr Thr Ser Leu Gln Glu Val Gln Asp340 345 350Ser Asp Asp Ala Asp Glu Ile Gln Ile Glu Leu Glu Asp Ala Ser Asp355 360 365Ala Trp Leu Val Val Ala Gln Glu Ala Arg Asp Phe Thr Leu Asn Ala370 375 380Tyr Ser Thr Asn Ser Arg Gln Asn Leu Pro Ile Asn Val Ile Ser Asp385 390 395 400Ser Cys Asn Cys Ser Thr Thr Asn Met Thr Ser Asn Gln Tyr Ser Asn405 410 415Pro Thr Thr Asn Met Thr Ser Asn Gln Tyr Met Ile Ser His Glu Tyr420 425 430Thr Ser Leu Pro Asn Asn Phe Met Leu Ser Arg Asn Ser Asn Leu Glu435 440 445Tyr Lys Cys Pro Glu Asn Asn Phe Met Ile Tyr Trp Tyr Asn Asn Ser450 455 460Asp Trp Tyr Asn Asn Ser Asp Trp Tyr Asn Asn465 470 475<210>41<211>1138<212>PRT<213>Bacillus thuringiensi<400>41Met Asn Leu Asn Asn Leu Asp Gly Tyr Glu Asp Ser Asn Arg Thr Leu1 5 10 15Asn Asn Ser Leu Asn Tyr Pro Thr Gln Lys Ala Leu Ser Pro Ser Leu20 25 30Lys Asn Met Asn Tyr Gln Asp Phe Leu Ser Ile Thr Glu Arg Glu Gln
35 40 45Pro Glu Ala Leu Ala Ser Gly Asn Thr Ala Ile Asn Thr Val Val Ser50 55 60Val Thr Gly Ala Thr Leu Ser Ala Leu Gly Val Pro Gly Ala Ser Phe65 70 75 80Ile Thr Asn Phe Tyr Leu Lys Ile Ala Gly Leu Leu Trp Pro Glu Asn85 90 95Gly Lys Ile Trp Asp Glu Phe Met Thr Glu Val Glu Ala Leu Ile Asp100 105 110Gln Lys Ile Glu Glu Tyr Val Arg Asn Lys Ala Ile Ala Glu Leu Asp115 120 125Gly Leu Gly Ser Ala Leu Asp Lys Tyr Gln Lys Ala Leu Ala Asp Trp130 135 140Leu Gly Lys Gln Asp Asp Pro Glu Ala Ile Leu Ser Val Ala Thr Glu145 150 155 160Phe Arg Ile Ile Asp Ser Leu Phe Glu Phe Ser Met Pro Ser Phe Lys165 170 175Val Thr Gly Tyr Glu Ile Pro Leu Leu Thr Val Tyr Ala Gln Ala Ala180 185 190Asn Leu His Leu Ala Leu Leu Arg Asp Ser Thr Leu Tyr Gly Asp Lys195 200 205Trp Gly Phe Thr Gln Asn Asn Ile Glu Glu Asn Tyr Asn Arg Gln Lys210 215 220Lys Arg Ile Ser Glu Tyr Ser Asp His Cys Thr Lys Trp Tyr Asn Ser225 230 235 240Gly Leu Ser Arg Leu Asn Gly Ser Thr Tyr Glu Gln Trp Ile Asn Tyr245 250 255Asn Arg Phe Arg Arg Glu Met Ile Leu Met Ala Leu Asp Leu Val Ala260 265 270Val Phe Pro Phe His Asp Pro Arg Arg Tyr Ser Met Glu Thr Ser Thr275 280 285Gln Leu Thr Arg Glu Val Tyr Thr Asp Pro Val Ser Leu Ser Ile Ser290 295 300Asn Pro Asp Ile Gly Pro Ser Phe Ser Gln Met Glu Asn Thr Ala Ile305 310 315 320Arg Thr Pro His Leu Val Asp Tyr Leu Asp Glu Leu Tyr Ile Tyr Thr325 330 335Ser Lys Tyr Lys Ala Phe Ser His Glu Ile Gln Pro Asp Leu Phe Tyr340 345 350Trp Ser Ala His Lys Val Ser Phe Lys Lys Ser Glu Gln Ser Asn Leu355 360 365Tyr Thr Thr Gly Ile Tyr Gly Lys Thr Ser Gly Tyr Ile Ser Ser Gly370 375 380Ala Tyr Ser Phe His Gly Asn Asp Ile Tyr Arg Thr Leu Ala Ala Pro385 390 395 400Ser Val Val Val Tyr Pro Tyr Thr Gln Asn Tyr Gly Val Glu Gln Val405 410 415Glu Phe Tyr Gly Val Lys Gly His Val His Tyr Arg Gly Asp Asn Lys420 425 430Tyr Asp Leu Thr Tyr Asp Ser Ile Asp Gln Leu Pro Pro Asp Gly Glu435 440 445Pro Ile His Glu Lys Tyr Thr His Arg Leu Cys His Ala Thr Ala Ile450 455 460Phe Lys Ser Thr Pro Asp Tyr Asp Asn Ala Thr Ile Pro Ile Phe Ser465 470 475 480Trp Thr His Arg Ser Ala Glu Tyr Tyr Asn Arg Ile Tyr Pro Asn Lys485 490 495Ile Thr Lys Ile Pro Ala Val Lys Met Tyr Lys Leu Asp Asp Pro Ser500 505 510Thr Val Val Lys Gly Pro Gly Phe Thr Gly Gly Asp Leu Val Lys Arg515 520 525Gly Ser Thr Gly Tyr Ile Gly Asp Ile Lys Ala Thr Val Asn Ser Pro530 535 540Leu Ser Gln Lys Tyr Arg Val Arg Val Arg Tyr Ala Thr Asn Val Ser545 550 555 560Gly Gln Phe Asn Val Tyr Ile Asn Asp Lys Ile Thr Leu Gln Thr Lys565 570 575Phe Gln Asn Thr Val Glu Thr Ile Gly Glu Gly Lys Asp Leu Thr Tyr
580 585 590Gly Ser Phe Gly Tyr Ile Glu Tyr Ser Thr Thr Ile Gln Phe Pro Asp595 600 605Glu His Pro Lys Ile Thr Leu His Leu Ser Asp Leu Ser Asn Asn Ser610 615 620Ser Phe Tyr Val Asp Ser Ile Glu Phe Ile Pro Val Asp Val Asn Tyr625 630 635 640Ala Glu Lys Glu Lys Leu Glu Lys Ala Gln Lys Ala Val Asn Thr Leu645 650 655Phe Thr Glu Gly Arg Asn Ala Leu Gln Lys Asp Val Thr Asp Tyr Lys660 665 670Val Asp Gln Val Ser Ile Leu Val Asp Cys Ile Ser Gly Asp Leu Tyr675 680 685Pro Asn Glu Lys Arg Glu Leu Gln Asn Leu Val Lys Tyr Ala Lys Arg690 695 700Leu Ser Tyr Ser Arg Asn Leu Leu Leu Asp Pro Thr Phe Asp Ser Ile705 710 715 720Asn Ser Ser Glu Glu Asn Gly Trp Tyr Gly Ser Asn Gly Ile Val Ile725 730 735Gly Asn Gly Asp Phe Val Phe Lys Gly Asn Tyr Leu Ile Phe Ser Gly740 745 750Thr Asn Asp Thr Gln Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu755 760 765Ser Lys Leu Lys Glu Tyr Thr Arg Tyr Lys Leu Lys Gly Phe Ile Glu770 775 780Ser Ser Gln Asp Leu Glu Ala Tyr Val Ile Arg Tyr Asp Ala Lys His785 790 795 800Arg Thr Leu Asp Val Ser Asp Asn Leu Leu Pro Asp Ile Leu Pro Glu805 810 815Asn Thr Cys Gly Glu Pro Asn Arg Cys Ala Ala Gln Gln Tyr Leu Asp820 825 830Glu Asn Pro Ser Pro Glu Cys Ser Ser Met Gln Asp Gly Ile Leu Ser835 840 845Asp Ser His Ser Phe Ser Leu Asn Ile Asp Thr Gly Ser Ile Asn His850 855 860Asn Glu Asn Leu Gly Ile Trp Val Leu Phe Lys Ile Ser Thr Leu Glu865 870 875 880Gly Tyr Ala Lys Phe Gly Asn Leu Glu Val Ile Glu Asp Gly Pro Val885 890 895Ile Gly Glu Ala Leu Ala Arg Val Lys Arg Gln Glu Thr Lys Trp Arg900 905 910Asn Lys Leu Ala Gln Leu Thr Thr Glu Thr Gln Ala Ile Tyr Thr Arg915 920 925Ala Lys Gln Ala Leu Asp Asn Leu Phe Ala Asn Ala Gln Asp Ser His930 935 940Leu Lys Arg Asp Val Thr Phe Ala Glu Ile Ala Ala Ala Arg Lys Ile945 950 955 960Val Gln Ser Ile Arg Glu Ala Tyr Met Ser Trp Leu Ser Val Val Pro965 970 975Gly Val Asn His Pro Ile Phe Thr Glu Leu Ser Gly Arg Val Gln Arg980 985 990Ala Phe Gln Leu Tyr Asp Val Arg Asn Val Val Arg Asn Gly Arg Phe995 10001005Leu Asn Gly Leu Ser Asp Trp Ile Val Thr Ser Asp Val Lys Val Gln101010151020Glu Glu Asn Gly Asn Asn Val Leu Val Leu Asn Asn Trp Asp Ala Gln102510301035 1040Val Leu Gln Asn Val Lys Leu Tyr Gln Asp Arg Gly Tyr Ile Leu His104510501055Val Thr Ala Arg Lys Ile Gly Ile Gly Glu Gly Tyr Ile Thr Ile Thr106010651070Asp Glu Glu Gly His Thr Asp Gln Leu Arg Phe Thr Ala Cys Glu Glu107510801085Ile Asp Ala Ser Asn Ala Phe Ile Ser Gly Tyr Ile Thr Lys Glu Leu109010951100Glu Phe Phe Pro Asp Thr Glu Lys Val His Ile Glu Ile Gly Glu Thr110511101115 1120Glu Gly Ile Phe Leu Val Glu Ser Ile Glu Leu Phe Leu Met Glu Glu
112511301135Leu Cys<210>42<211>1157<212>PRT<213>Bacillus thuringiensi<400>42Met Ser Pro Asn Asn Gln Asn Glu Tyr Glu Ile Ile Asp Ala Thr Pro1 5 10 15Ser Thr Ser Val Ser Ser Asp Ser Asn Arg Tyr Pro Phe Ala Asn Glu20 25 30Pro Thr Asp Ala Leu Gln Asn Met Asn Tyr Lys Asp Tyr Leu Lys Met35 40 45Ser Gly Gly Glu Asn Pro Glu Leu Phe Gly Asn Pro Glu Thr Phe Ile50 55 60Ser Ser Ser Thr Ile Gln Thr Gly Ile Gly Ile Val Gly Arg Ile Leu65 70 75 80Gly Ala Leu Gly Val Pro Phe Ala Ser Gln Ile Ala Ser Phe Tyr Ser85 90 95Phe Ile Val Gly Gln Leu Trp Pro Ser Lys Ser Val Asp Ile Trp Gly100 105 110Glu Ile Met Glu Arg Val Glu Glu Leu Val Asp Gln Lys Ile Glu Lys115 120 125Tyr Val Lys Asp Lys Ala Leu Ala Glu Leu Lys Gly Leu Gly Asn Ala130 135 140Leu Asp Val Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn Arg Asn145 150 155 160Asp Ala Arg Thr Arg Ser Val Val Ser Asn Gln Phe Ile Ala Leu Asp165 170 175Leu Asn Phe Val Ser Ser Ile Pro Ser Phe Ala Val Ser Gly His Glu180 185 190Val Leu Leu Leu Ala Val Tyr Ala Gln Ala Val Asn Leu His Leu Leu195 200 205Leu Leu Arg Asp Ala Ser Ile Phe Gly Glu Glu Trp Gly Phe Thr Pro210 215 220Gly Glu Ile Ser Arg Phe Tyr Asn Arg Gln Val Gln Leu Thr Ala Glu225 230 235 240Tyr Ser Asp Tyr Cys Val Lys Trp Tyr Lys Ile Gly Leu Asp Lys Leu245 250 255Lys Gly Thr Thr Ser Lys Ser Trp Leu Asn Tyr His Gln Phe Arg Arg260 265 270Glu Met Thr Leu Leu Val Leu Asp Leu Val Ala Leu Phe Pro Asn Tyr275 280 285Asp Thr His Met Tyr Pro Ile Glu Thr Thr Ala Gln Leu Thr Arg Asp290 295 300Val Tyr Thr Asp Pro Ile Ala Phe Asn Ile Val Thr Ser Thr Gly Phe305 310 315 320Cys Asn Pro Trp Ser Thr His Ser Gly Ile Leu Phe Tyr Glu Val Glu325 330 335Asn Asn Val Ile Arg Pro Pro His Leu Phe Asp Ile Leu Ser Ser Val340 345 350Glu Ile Asn Thr Ser Arg Gly Gly Ile Thr Leu Asn Asn Asp Ala Tyr355 360 365Ile Asn Tyr Trp Ser Gly His Thr Leu Lys Tyr Arg Arg Thr Ala Asp370 375 380Ser Thr Val Thr Tyr Thr Ala Asn Tyr Gly Arg Ile Thr Ser Glu Lys385 390 395 400Asn Ser Phe Ala Leu Glu Asp Arg Asp Ile Phe Glu Ile Asn Ser Thr405 410 415Val Ala Asn Leu Ala Asn Tyr Tyr Gln Lys Ala Tyr Gly Val Pro Gly420 425 430Ser Trp Phe His Met Val Lys Arg Gly Thr Ser Ser Thr Thr Ala Tyr435 440 445Leu Tyr Ser Lys Thr His Thr Ala Leu Gln Gly Cys Thr Gln Val Tyr
450 455 460Glu Ser Ser Asp Glu Ile Pro Leu Asp Arg Thr Val Pro Val Ala Glu465 470 475 480Ser Tyr Ser His Arg Leu Ser His Ile Thr Ser His Ser Phe Ser Lys485 490 495Asn Gly Ser Ala Tyr Tyr Gly Ser Phe Pro Val Phe Val Trp Thr His500 505 510Thr Ser Ala Asp Leu Asn Asn Thr Ile Tyr Ser Asp Lys Ile Thr Gln515 520 525Ile Pro Ala Val Lys Gly Asp Met Leu Tyr Leu Gly Gly Ser Val Val530 535 540Gln Gly Pro Gly Phe Thr Gly Gly Asp Ile Leu Lys Arg Thr Asn Pro545 550 555 560Ser Ile Leu Gly Thr Phe Ala Val Thr Val Asn Gly Ser Leu Ser Gln565 570 575Arg Tyr Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asp Phe Glu Phe580 585 590Thr Leu Tyr Leu Gly Asp Thr Ile Glu Lys Asn Arg Phe Asn Lys Thr595 600 605Met Asp Asn Gly Ala Ser Leu Thr Tyr Glu Thr Phe Lys Phe Ala Ser610 615 620Phe Ile Thr Asp Phe Gln Phe Arg Glu Thr Gln Asp Lys Ile Leu Leu625 630 635 640Ser Met Gly Asp Phe Ser Ser Gly Gln Glu Val Tyr Ile Asp Arg Ile645 650 655Glu Phe Ile Pro Val Asp Glu Thr Tyr Glu Ala Glu Gln Asp Leu Glu660 665 670Ala Ala Lys Lys Ala Val Asn Ala Leu Phe Thr Asn Thr Lys Asp Gly675 680 685Leu Arg Pro Gly Val Thr Asp Tyr Glu Val Asn Gln Ala Ala Asn Leu690 695 700Val Glu Cys Leu Ser Asp Asp Leu Tyr Pro Asn Glu Lys Arg Leu Leu705 710 715 720Phe Asp Ala Val Arg Glu Ala Lys Arg Leu Ser Gly Ala Arg Asn Leu725 730 735Leu Gln Asp Pro Asp Phe Gln Glu Ile Asn Gly Glu Asn Gly Trp Ala740 745 750Ala Ser Thr Gly Ile Glu Ile Val Glu Gly Asp Ala Val Phe Lys Gly755 760 765Arg Tyr Leu Arg Leu Pro Gly Ala Arg Glu Ile Asp Thr Glu Thr Tyr770 775 780Pro Thr Tyr Leu Tyr Gln Lys Val Glu Glu Gly Val Leu Lys Pro Tyr785 790 795 800Thr Arg Tyr Arg Leu Arg Gly Phe Val Gly Ser Ser Gln Gly Leu Glu805 810 815Ile Tyr Thr Ile Arg His Gln Thr Asn Arg Ile Val Lys Asn Val Pro820 825 830Asp Asp Leu Leu Pro Asp Val Ser Pro Val Asn Ser Asp Gly Ser Ile835 840 845Asn Arg Cys Ser Glu Gln Lys Tyr Val Asn Ser Arg Leu Glu Gly Glu850 855 860Asn Arg Ser Gly Asp Ala His Glu Phe Ser Leu Pro Ile Asp Ile Gly865 870 875 880Glu Leu Asp Tyr Asn Glu Asn Ala Gly Ile Trp Val Gly Phe Lys Ile885 890 895Thr Asp Pro Glu Gly Tyr Ala Thr Leu Gly Asn Leu Glu Leu Val Glu900 905 910Glu Gly Pro Leu Ser Gly Asp Ala Leu Glu Arg Leu Gln Arg Glu Glu915 920 925Gln Gln Trp Lys Ile Gln Met Thr Arg Arg Arg Glu Glu Thr Asp Arg930 935 940Arg Tyr Met Ala Ser Lys Gln Ala Val Asp Arg Leu Tyr Ala Asp Tyr945 950 955 960Gln Asp Gln Gln Leu Asn Pro Asp Val Glu Ile Thr Asp Leu Thr Ala965 970 975Ala Gln Asp Leu Ile Gln Ser Ile Pro Tyr Val Tyr Asn Glu Met Phe980 985 990Pro Glu Ile Pro Gly Met Asn Tyr Thr Lys Phe Thr Glu Leu Thr Asp
995 10001005Arg Leu Gln Gln Ala Trp Asn Leu Tyr Asp Gln Arg Asn Ala Ile Pro101010151020Asn Gly Asp Phe Arg Asn Gly Leu Ser Asn Trp Asn Ala Thr Pro Gly102510301035 1040Val Glu Val Gln Gln Ile Asn His Thr Ser Val Leu Val Ile Pro Asn104510501055Trp Asp Glu Gln Val Ser Gln Gln Phe Thr Val Gln Pro Asn Gln Arg106010651070Tyr Val Leu Arg Val Thr Ala Arg Lys Glu Gly Val Gly Asn Gly Tyr107510801085Val Ser Ile Arg Asp Gly Gly Asn Gln Ser Glu Thr Leu Thr Phe Ser109010951100Ala Ser Asp Tyr Asp Thr Asn Gly Val Tyr Asn Asp Gln Thr Gly Tyr110511101115 1120Ile Thr Lys Thr Val Thr Phe Ile Pro Tyr Thr Asp Gln Met Trp Ile112511301135Glu Ile Ser Glu Thr Glu Gly Thr Phe Tyr Ile Glu Ser Val Glu Leu114011451150Ile Val Asp Val Glu1155<210>43<211>675<212>PRT<213>Bacillus thuringiensi<400>43Met Asn Pro Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Phe Asn Ala Pro1 5 10 15Ser Asn Gly Phe Ser Lys Ser Asn Asn Tyr Ser Arg Tyr Pro Leu Ala20 25 30Asn Lys Pro Asn Gln Pro Leu Lys Asn Thr Asn Tyr Lys Asp Trp Leu35 40 45Asn Val Cys Gln Asp Asn Gln Gln Tyr Gly Asn Asn Ala Gly Asn Phe50 55 60Ala Ser Ser Glu Thr Ile Val Gly Val Ser Ala Gly Ile Ile Val Val65 70 75 80Gly Thr Met Leu Gly Ala Phe Ala Ala Pro Val Leu Ala Ala Gly Ile85 90 95Ile Ser Phe Gly Thr Leu Leu Pro Ile Phe Trp Gln Gly Ser Asp Pro100 105 110Ala Asn Val Trp Gln Asp Leu Leu Asn Ile Gly Gly Arg Pro Ile Gln115 120 125Glu Ile Asp Lys Asn Ile Ile Asn Val Leu Thr Ser Ile Val Thr Pro130 135 140Ile Lys Asn Gln Leu Asp Lys Tyr Gln Glu Phe Phe Asp Lys Trp Glu145 150 155 160Pro Ala Arg Thr His Ala Asn Ala Lys Ala Val His Asp Leu Phe Thr165 170 175Thr Leu Glu Pro Ile Ile Asp Lys Asp Leu Asp Met Leu Lys Asn Asn180 185 190Ala Ser Tyr Arg Ile Pro Thr Leu Pro Ala Tyr Ala Gln Ile Ala Thr195 200 205Trp His Leu Asn Leu Leu Lys His Ala Ala Thr Tyr Tyr Asn Ile Trp210 215 220Leu Gln Asn Gln Gly Ile Asn Pro Ser Thr Phe Asn Ser Ser Asn Tyr225 230 235 240Tyr Gln Gly Tyr Leu Lys Arg Lys Ile Gln Glu Tyr Thr Asp Tyr Cys245 250 255Ile Gln Thr Tyr Asn Ala Gly Leu Thr Met Ile Arg Thr Asn Thr Asn260 265 270Ala Thr Trp Asn Met Tyr Asn Thr Tyr Arg Leu Glu Met Thr Leu Thr275 280 285Val Leu Asp Leu Ile Ala Ile Phe Pro Asn Tyr Asp Pro Glu Lys Tyr290 295 300Pro Ile Gly Val Lys Ser Glu Leu Ile Arg Glu Val Tyr Thr Asn Val
305 310 315 320Asn Ser Asp Thr Phe Arg Thr Ile Thr Glu Leu Glu Asn Gly Leu Thr325 330 335Arg Asn Pro Thr Leu Phe Thr Trp Ile Asn Gln Gly Arg Phe Tyr Thr340 345 350Arg Asn Ser Arg Asp Ile Leu Asp Pro Tyr Asp Ile Phe Ser Phe Thr355 360 365Gly Asn Gln Met Ala Phe Thr His Thr Asn Asp Asp Arg Ash Ile Ile370 375 380Trp Gly Ala Val His Gly Asn Ile Ile Ser Gln Asp Thr Ser Lys Val385 390 395 400Phe Pro Phe Tyr Arg Asn Lys Pro Ile Asp Lys Val Glu Ile Val Arg405 410 415His Arg Glu Tyr Ser Asp Ile Ile Tyr Glu Met Ile Phe Phe Ser Asn420 425 430Ser Ser Glu Val Phe Arg Tyr Ser Ser Asn Ser Thr Ile Glu Asn Asn435 440 445Tyr Lys Arg Thr Asp Ser Tyr Met Ile Pro Lys Gln Thr Trp Lys Asn450 455 460Glu Glu Tyr Gly His Thr Leu Ser Tyr Ile Lys Thr Asp Asn Tyr Ile465 470 475 480Phe Ser Val Val Arg Glu Arg Arg Arg Val Ala Phe Ser Trp Thr His485 490 495Thr Ser Val Asp Phe Gln Asn Thr Ile Asp Leu Asp Asn Ile Thr Gln500 505 510Ile His Ala Leu Lys Ala Leu Lys Val Ser Ser Asp Ser Lys Ile Val515 520 525Lys Gly Pro Gly His Thr Gly Gly Asp Leu Val Ile Leu Lys Asp Ser530 535 540Met Asp Phe Arg Val Arg Phe Leu Lys Asn Val Ser Arg Gln Tyr Gln545 550 555 560Val Arg Ile Arg Tyr Ala Thr Asn Ala Pro Lys Thr Thr Val Phe Leu565 570 575Thr Gly Ile Asp Thr Ile Ser Val Glu Leu Pro Ser Thr Thr Ser Arg580 585 590Gln Asn Pro Asn Ala Thr Asp Leu Thr Tyr Ala Asp Phe Gly Tyr Val595 600 605Thr Phe Pro Arg Thr Val Pro Asn Lys Thr Phe Glu Gly Glu Asp Thr610 615 620Leu Leu Met Thr Leu Tyr Gly Thr Pro Asn His Ser Tyr Asn Ile Tyr625 630 635 640Ile Asp Lys Ile Glu Phe Ile Pro Ile Thr Gln Ser Val Leu Asp Tyr645 650 655Thr Glu Lys Gln Asn Ile Glu Lys Thr Gln Lys Ile Val Asn Asp Leu660 665 670Phe Val Asn675<210>44<211>648<212>PRT<213>Bacillus thuringiensi<400>44Met His Tyr Tyr Gly Asn Arg Asn Glu Tyr Asp Ile Leu Asn Ala Ser1 5 10 15Ser Asn Asp Ser Asn Met Ser Asn Thr Tyr Pro Arg Tyr Pro Leu Ala20 25 30Asn Pro Gln Gln Asp Leu Met Gln Asn Thr Asn Tyr Lys Asp Trp Leu35 40 45Asn Val Cys Glu Gly Tyr His Ile Glu Asn Pro Arg Glu Ala Ser Val50 55 60Arg Ala Gly Leu Gly Lys Gly Leu Gly Ile Val Ser Thr Ile Val Gly65 70 75 80Phe Phe Gly Gly Ser Ile Ile Leu Asp Thr Ile Gly Leu Phe Tyr Gln85 90 95Ile Ser Glu Leu Leu Trp Pro Glu Asp Asp Thr Gln Gln Tyr Thr Trp
100 105 110Gln Asp Ile Met Asn His Val Glu Asp Leu Ile Asp Lys Arg Ile Thr115 120 125Glu Val Ile Arg Gly Asn Ala Ile Arg Thr Leu Ala Asp Leu Gln Gly130 135 140Lys Val Asp Asp Tyr Asn Asn Trp Leu Lys Lys Trp Lys Asp Asp Pro145 150 155 160Lys Ser Thr Gly Asn Leu Ser Thr Leu Val Thr Lys Phe Thr Ala Leu165 170 175Asp Ser Asp Phe Asn Gly Ala Ile Arg Thr Val Asn Asn Gln Gly Ser180 185 190Pro Gly Tyr Glu Leu Leu Leu Leu Pro Val Tyr Ala Gln Ile Ala Asn195 200 205Leu His Leu Leu Leu Leu Arg Asp Ala Gln Ile Tyr Gly Asp Lys Trp210 215 220Trp Ser Ala Arg Ala Asn Ala Arg Asp Asn Tyr Tyr Gln Ile Gln Leu225 230 235 240Glu Lys Thr Lys Glu Tyr Thr Glu Tyr Cys Ile Asn Trp Tyr Asn Lys245 250 255Gly Leu Asn Asp Phe Arg Thr Ala Gly Gln Trp Val Asn Phe Asn Arg260 265 270Tyr Arg Arg Glu Met Thr Leu Thr Val Leu Asp Ile Ile Ser Met Phe275 280 285Pro Ile Tyr Asp Ala Arg Leu TyT Pro Thr Glu Val Lys Thr Glu Leu290 295 300Thr Arg Glu Ile Tyr Ser Asp Val Ile Asn Gly Glu Ile Tyr Gly Leu305 310 315 320Met Thr Pro Tyr Phe Ser Phe Glu Lys Ala Glu Ser Leu Tyr Thr Arg325 330 335Ala Pro His Leu Phe Thr Trp Leu Lys Gly Phe Arg Phe Val Thr Asn340 345 350Ser Ile Ser Tyr Trp Thr Phe Leu Ser Gly Gly Gln Asn Lys Tyr Ser355 360 365Tyr Thr Asn Asn Ser Ser Ile Asn Glu Gly Ser Phe Arg Gly Gln Asp370 375 380Thr Asp Tyr Gly Gly Thr Ser Ser Thr Ile Asn Ile Pro Ser Asn Ser385 390 395 400Tyr Val Tyr Asn Leu Trp Thr Glu Asn Tyr Glu Tyr Ile Tyr Pro Trp405 410 415Gly Asp Pro Val Asn Ile Thr Lys Met Asn Phe Ser Val Thr Asp Asn420 425 430Asn Ser Ser Lys Glu Leu Ile Tyr Gly Ala His Arg Thr Asn Lys Pro435 440 445Val Val Arg Thr Asp Phe Asp Phe Leu Thr Asn Lys Glu Gly Thr Glu450 455 460Leu Ala Lys Tyr Asn Asp Tyr Asn His Ile Leu Ser Tyr Met Leu Ile465 470 475 480Asn Gly Glu Thr Phe Gly Gln Lys Arg His Gly Tyr Ser Phe Ala Phe485 490 495Thr His Ser Ser Val Asp Pro Asn Asn Thr Ile Ala Ala Asn Lys Ile500 505 510Thr Gln Ile Pro Val Val Lys Ala Ser Ser Ile Asn Gly Ser Ile Ser515 520 525Ile Glu Lys Gly Pro Gly Phe Thr Gly Gly Asp Leu Val Lys Met Arg530 535 540Ala Asp Ser Gly Leu Thr Met Arg Phe Lys Ala Glu Leu Leu Asp Lys545 550 555 560Lys Tyr Arg Val Arg Ile Arg Tyr Lys Cys Asn Tyr Ser Ser Lys Leu565 570 575Ile Leu Arg Lys Trp Lys Gly Glu Gly Tyr Ile Gln Gln Gln Ile His580 585 590Asn Ile Ser Pro Thr Tyr Gly Ala Phe Ser Tyr Leu Glu Ser Phe Thr595 600 605Ile Thr Thr Thr Glu Asn Ile Phe Asp Leu Thr Met Glu Val Thr Tyr610 615 620Pro Tyr Gly Arg Gln Phe Val Glu Asp Ile Pro Ser Leu Ile Leu Asp625 630 635 640Lys Ile Glu Phe Leu Pro Thr Asn
645<210>45<211>682<212>PRT<213>Bacillus thuringiensi<400>45Met Asn Ser Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Leu Asp Ala Lys1 5 10 15Arg Asn Thr Cys His Met Ser Asn Cys Tyr Pro Lys Tyr Pro Leu Ala20 25 30Asn Asp Pro Gln Met Tyr Leu Arg Asn Thr His Tyr Lys Asp Trp Ile35 40 45Asn Met Cys Glu Glu Ala Ser Tyr Ala Ser Ser Gly Pro Ser Gln Leu50 55 60Phe Lys Val Gly Gly Ser Ile Val Ala Lys Ile Leu Gly Met Ile Pro65 70 75 80Glu Val Gly Pro Leu Leu Ser Trp Met Val Ser Leu Phe Trp Pro Thr85 90 95Ile Glu Glu Lys Asn Thr Val Trp Glu Asp Met Ile Lys Tyr Val Ala100 105 110Asn Leu Leu Lys Gln Glu Leu Thr Asn Asp Thr Leu Asn Arg Ala Thr115 120 125Ser Asn Leu Ser Gly Leu Asn Glu Ser Leu Asn Ile Tyr Asn Arg Ala130 135 140Leu Ala Ala Trp Lys Gln Asn Lys Asn Asn Phe Ala Ser Gly Glu Leu145 150 155 160Ile Arg Ser Tyr Ile Asn Asp Leu His Ile Leu Phe Thr Arg Asp Ile165 170 175Gln Ser Asp Phe Ser Leu Gly Gly Tyr Glu Thr Val Leu Leu Pro Ser180 185 190Tyr Ala Ser Ala Ala Asn Leu His Leu Leu Leu Leu Arg Asp Val Ala195 200 205Ile Tyr Gly Lys Glu Leu Gly Tyr Pro Ser Thr Asp Val Glu Phe Tyr210 215 220Tyr Asn Glu Gln Lys Tyr Tyr Thr Glu Lys Tyr Ser Asn Tyr Cys Val225 230 235 240Asn Thr Tyr Lys Ser Gly Leu Glu Ser Lys Lys Gln Ile Gly Trp Ser245 250 255Asp Phe Asn Arg Tyr Arg Arg Glu Met Thr Leu Ser Val Leu Asp Ile260 265 270Val Ala Leu Phe Pro Leu Tyr Asp Thr Gly Leu Tyr Pro Ser Lys Asp275 280 285Gly Lys Ile His Val Lys Ala Glu Leu Thr Arg Glu Ile Tyr Ser Asp290 295 300Val Ile Asn Asp His Val Tyr Gly Leu Met Val Pro Tyr Ile Ser Phe305 310 315 320Glu His Ala Glu Ser Leu Tyr Thr Arg Arg Pro His Ala Phe Thr Trp325 330 335Leu Lys Gly Phe Arg Phe Val Thr Asn Ser Ile Asn Ser Trp Thr Phe340 345 350Leu Ser Gly Gly Glu Asn Arg Tyr Phe Leu Thr His Gly Glu Gly Thr355 360 365Ile Tyr Asn Gly Pro Phe Leu Gly Gln Asp Thr Glu Tyr Gly Gly Thr370 375 380Ser Ser Tyr Ile Asp Ile Ser Asn Asn Ser Ser Ile Tyr Asn Leu Trp385 390 395 400Thr Lys Asn Tyr Glu Trp Ile Tyr Pro Trp Thr Asp Pro Val Asn Ile405 410 415Thr Lys Ile Asn Phe Ser Ile Thr Asp Ash Ser Asn Ser Ser Glu Ser420 425 430Ile Tyr Gly Ala Glu Arg Met Asn Lys Pro Thr Val Arg Thr Asp Phe435 440 445Asn Phe Leu Leu Asn Arg Ala Gly Asn Gly Pro Thr Thr Tyr Asn Asp450 455 460Tyr Asn His Ile Leu Ser Tyr Met Leu Ile Asn Gly Glu Thr Phe Gly
465 470 475 480Gln Lys Arg His Gly Tyr Ser Phe Ala Phe Thr His Ser Ser Val Asp485 490 495Arg Tyr Asn Thr Ile Val Pro Asp Lys Ile Val Gln Ile Pro Ala Val500 505 510Lys Thr Asn Leu Val Gly Ala Asn Ile Ile Lys Gly Pro Gly His Thr515 520 525Gly Gly Asp Leu Leu Lys Leu Glu Tyr Glu Arg Phe Leu Ser Leu Arg530 535 540Ile Lys Leu Ile Ala Ser Met Thr Phe Arg Ile Arg Ile Arg Tyr Ala545 550 555 560Ser Asn Ile Ser Gly Gln Met Met Ile Asn Ile Gly Tyr Gln Asn Pro565 570 575Thr Tyr Phe Asn Ile Ile Pro Thr Thr Ser Arg Asp Tyr Thr Glu Leu580 585 590Lys Phe Glu Asp Phe Gln Leu Val Asp Thr Ser Tyr Ile Tyr Ser Gly595 600 605Gly Pro Ser Ile Ser Ser Asn Thr Leu Trp Leu Asp Asn Phe Ser Asn610 615 620Gly Pro Val Ile Ile Asp Lys Ile Glu Phe Ile Pro Leu Gly Ile Thr625 630 635 640Leu Asn Gln Ala Gln Gly Tyr Asp Thr Tyr Asp Gln Asn Ala Asn Gly645 650 655Met Tyr His Gln Asn Tyr Ser Asn Ser Gly Tyr Asn Tyr Asn Gln Glu660 665 670Tyr Asn Thr Tyr Tyr Gln Ser Tyr Asn Asn675 680<210>46<211>529<212>PRT<213>Bacillus thuringiensi<400>46Val Asn Phe Met Leu Thr Ser Gly Ala Lys Asn Met Leu Lys Leu Glu1 5 10 15Thr Thr Asp Tyr Glu Ile Asp Gln Met Ala Asn Ala Ile Glu Asn Met20 25 30Ser Gly Glu Gln Tyr Ser Gln Glu Lys Met Met Gln Trp His Asp Ile35 40 45Lys Tyr Ala Lys Gln Leu Ser Gln Ala Arg Asn Leu Leu Gln Asn Gly50 55 60Asp Phe Glu Asp Leu Phe Ser Gly Trp Thr Thr Ser Asn Gln Met Ser65 70 75 80Ile Gln Ala Asp Asn Ala Thr Phe Lys Gly Asn Tyr Leu His Met Ser85 90 95Gly Ala Arg Asp Ile Tyr Gly Thr Ile Phe Pro Thr Tyr Ile Tyr Gln100 105 110Lys Ile Asp Glu Ser Lys Leu Lys Pro Tyr Thr Arg Tyr Leu Val Arg115 120 125Gly Phe Val Gly Ser Ser Lys Asp Leu Glu Leu Met Val Met Arg Tyr130 135 140Gly Lys Glu Ile Asp Thr Val Met Asn Val Pro Asn Asp Ile Pro Tyr145 150 155 160Val Pro Ser Met Pro Val Cys Asn Glu Leu Tyr Asp Gly Gln Gln Pro165 170 175Tyr Pro Asn Arg His Val Gly Tyr Tyr Asn Pro Met Pro Val Ser Gln180 185 190Pro Ser Tyr Thr Ser Asp Thr Cys Gln Cys Thr Pro Gly Lys Lys His195 200 205Val Val Cys His Asp Ser His Gln Phe Lys Phe His Ile Asp Thr Gly210 215 220Glu Val Asp Tyr Asn Thr Asn Leu Gly Ile Trp Val Leu Phe Lys Ile225 230 235 240Ser Ser Pro Asp Gly Tyr Ala Thr Leu Asp Asn Leu Glu Val Ile Glu245 250 255Glu Gly Pro Val Arg Gly Glu Ala Val Thr His Val Lys Gln Lys Glu
260 265 270Lys Lys Trp Asn Gln Gln Met Glu Lys Lys Arg Met Glu Thr Lys Arg275 280 285Val Tyr Asp Arg Ala Lys Gln Ala Val Asp Ala Leu Phe Thr Gly Glu290 295 300Glu Leu Asn Tyr Asp Val Thr Leu Ser His Ile Lys Asn Ala Asp Asp305 310 315 320Leu Val Gln Ser Ile Pro Tyr Val His Asn Glu Trp Leu Pro Asp Phe325 330 335Pro Gly Met Asn Tyr Asp Ile Tyr Gln Glu Leu Asn Ala Arg Ile Met340 345 350Gln Ala Arg Tyr Leu Tyr Asp Ala Arg Asn Val Ile Thr Asn Gly Asp355 360 365Phe Ala Gln Gly Leu Gln Gly Trp His Ala Glu Gly Lys Val Glu Val370 375 380Gln Gln Met Asn Gly Thr Ser Val Leu Val Leu Ser Asn Trp Ser Ser385 390 395 400Gly Val Ser Gln Asn Leu His Val Gln His Pro His Gly Tyr Leu Leu405 410 415Arg Val Ser Ala Lys Lys Glu Gly Ser Gly Lys Gly Tyr Val Thr Arg420 425 430Met Ser Cys Asn Gly Lys Gln Glu Thr Leu Thr Phe Thr Ser Cys Asp435 440 445Gly Gly Tyr Met Thr Lys Thr Val Glu Val Phe Pro Glu Ser Asp Arg450 455 460Val Arg Ile Glu Ile Gly Glu Thr Glu Gly Ser Phe Tyr Ile Glu Ser465 470 475 480Ile Glu Leu Ile Cys Met Asn Gly Tyr Thr Ser Asn Asn Asn Gln Asn485 490 495Met Ser Asn Met Tyr Asp Gln Ser Tyr Ser Gly Asn Tyr Ser Gln Asn500 505 510Thr Ser Asp Met Tyr Asp Gln Gly Gly Ser Val Ala Lys Phe Glu Lys515 520 525Glu<210>47<211>674<212>PRT<213>Bacillus thuringiensi<400>47Met Asn Gln Tyr Gln Asn Lys Asn Glu Tyr Glu Ile Leu Glu Ser Ser1 5 10 15Gln Asn Asn Met Asn Met Pro Asn Arg Tyr Pro Phe Ala Asp Asp Pro20 25 30Asn Ala Val Met Lys Asn Gly Asn Tyr Lys Asp Trp Val Asn Glu Cys35 40 45Glu Gly Ser Asn Ile Ser Pro Ser Pro Ala Ala Ala Ile Thr Ser Lys50 55 60Ile Val Ser Ile Val Leu Lys Thr Leu Ala Lys Ala Val Ala Ser Ser65 70 75 80Leu Ala Asp Ser Ile Lys Ser Ser Leu Gly Ile Ser Lys Thr Ile Thr85 90 95Glu Asn Asn Val Ser Gln Val Ser Met Val Gln Val His Gln Ile Ile100 105 110Asn Arg Arg Ile Gln Glu Thr Ile Leu Asp Leu Gly Glu Ser Ser Leu115 120 125Asn Gly Leu Val Ala Ile Tyr Asn Arg Asp Tyr Leu Gly Ala Leu Glu130 135 140Ala Trp Asn Asn Asn Lys Ser Asn Ile Asn Tyr Gln Thr Asn Val Ala145 150 155 160Glu Ala Phe Lys Thr Val Glu Arg Glu Phe Phe Thr Lys Leu Lys Gly165 170 175Ile Tyr Arg Thr Ser Ser Ser Gln Ile Thr Leu Leu Pro Thr Phe Thr180 185 190Gln Ala Ala Asn Leu His Leu Ser Met Leu Arg Asp Ala Val Met Tyr
195 200 205Gln Glu Gly Trp Asn Leu Gln Ser His Ile Asn Tyr Ser Lys Glu Leu210 215 220Asp Asp Ala Leu Glu Asp Tyr Thr Asn Tyr Cys Val Glu Val Tyr Thr225 230 235 240Lys Gly Leu Asn Ala Leu Arg Gly Ser Thr Ala Ile Asp Trp Leu Glu245 250 255Phe Asn Ser Phe Arg Arg Asp Met Thr Leu Met Val Leu Asp Leu Val260 265 270Ala Ile Phe Pro Asn Tyr Asn Pro Val Arg Tyr Pro Leu Ser Thr Lys275 280 285Ile Ser Leu Ser Arg Lys Ile Tyr Thr Asp Pro Val Gly Arg Thr Asp290 295 300Ser Pro Ser Phe Gly Asp Trp Thr Asn Thr Gly Arg Thr Leu Ala Asn305 310 315 320Phe Asn Asp Leu Glu Arg Glu Val Thr Asp Ser Pro Ser Leu Val Lys325 330 335Trp Leu Gly Asp Met Thr Ile Tyr Thr Gly Ala Ile Asp Ser Tyr Arg340 345 350Pro Thr Ser Pro Gly Asp Arg Ile Gly Val Trp Tyr Gly Asn Ile Asn355 360 365Ala Phe Tyr His Thr Gly Arg Thr Asp Val Val Met Phe Arg Gln Thr370 375 380Gly Asp Thr Ala Tyr Glu Asp Pro Ser Thr Phe Ile Ser Asn Ile Leu385 390 395 400Tyr Asp Asp Ile Tyr Lys Leu Asp Leu Arg Ala Ala Ala Val Ser Thr405 410 415Ile Gln Gly Ala Met Asp Thr Thr Phe Gly Val Ser Ser Ser Arg Phe420 425 430Phe Asp Ile Arg Gly Arg Asn Gln Leu Tyr Gln Ser Asn Lys Pro Tyr435 440 445Pro Ser Leu Pro Ile Thr Ile Thr Phe Pro Gly Glu Glu Ser Ser Glu450 455 460Gly Asn Ala Asn Asp Tyr Ser His Leu Leu Cys Asp Val Lys Ile Leu465 470 475 480Gln Glu Asp Ser Ser Asn Ile Cys Glu Gly Arg Ser Ser Leu Leu Ser485 490 495His Ala Trp Thr His Ala Ser Leu Asp Arg Asn Asn Thr Ile Leu Pro500 505 510Asp Glu Ile Thr Gln Ile Pro Ala Val Thr Ala Tyr Glu Leu Arg Gly515 520 525Asn Ser Ser Val Val Ala Gly Pro Gly Ser Thr Gly Gly Asp Leu Val530 535 540Lys Met Ser Tyr His Ser Val Trp Ser Phe Lys Val Tyr Cys Ser Glu545 550 555 560Leu Lys Asn Tyr Arg Val Arg Ile Arg Tyr Ala Ser His Gly Asn Cys565 570 575Gln Phe Leu Met Lys Arg Trp Pro Ser Thr Gly Val Ala Pro Arg Gln580 585 590Trp Ala Arg His Asn Val Gln Gly Thr Phe Ser Asn Ser Met Arg Tyr595 600 605Glu Ala Phe Lys Tyr Leu Asp Ile Phe Thr Ile Thr Pro Glu Glu Asn610 615 620Asn Phe Ala Phe Thr Ile Asp Leu Glu Ser Gly Gly Asp Leu Phe Ile625 630 635 640Asp Lys Ile Glu Phe Ile Pro Val Ser Gly Ser Ala Phe Glu Tyr Glu645 650 655Gly Lys Gln Asn Ile Glu Lys Thr Gln Lys Ala Val Asn Asp Leu Phe660 665 670Ile Asn<210>48<211>675<212>PRT<213>Bacillus thuringiensi
<400>48Met Asn Pro Tyr Gln Asn Lys Ser Glu Cys Glu Ile Leu Asn Ala Pro1 5 10 15Leu Asn Asn Ile Asn Met Pro Asn Arg Tyr Pro Phe Ala Asn Asp Pro20 25 30Asn Ala Val Met Lys Asn Gly Asn Tyr Lys Asp Trp Leu Asn Glu Cys35 40 45Asp Gly Ile Thr Pro Ser Ile Phe Gly Thr Leu Gly Val Leu Ala Ser50 55 60Ile Val Ile Ser Thr Ile Asn Leu Ala Thr Ser Pro Ser Ile Gly Asp65 70 75 80Ala Phe Ala Leu Val Ser Ser Ile Gly Glu Tyr Trp Pro Glu Thr Lys85 90 95Thr Ser Phe Pro Leu Ser Val Ala Asp Val Asn Arg Leu Ile Arg Glu100 105 110Ala Leu Asp Gln Asn Ala Ile Asn Arg Ala Thr Gly Lys Phe Asn Gly115 120 125Leu Met Asp Thr Tyr Asn Thr Val Tyr Leu Lys Asn Leu Gln Asp Trp130 135 140Tyr Asp Thr Arg Ile Pro Ala Asn Pro Gln Gly Asp Ser Gln Leu Arg145 150 155 160Glu Ala Ala Arg Arg Ser Leu Glu Glu Ile Glu Arg Asp Phe Arg Lys165 170 175Ala Leu Ala Gly Glu Phe Ala Glu Ala Gly Ser Gln Ile Val Leu Leu180 185 190Pro Ile Tyr Ala Gln Ala Ala Asn Ile His Leu Leu Ile Leu Lys Asp195 200 205Ala Met Gln Phe Arg Thr Asp Leu Gly Leu Ile Arg Pro Val Gly Val210 215 220Pro Ile Thr Thr Ser Ala Glu Asp Pro Phe Glu Ser Glu Phe Leu Leu225 230 235 240Arg Ile Lys Lys Tyr Thr Asp His Cys Ile Ser Tyr Tyr Asp Asp Gly245 250 255Leu Ala Lys Ile Arg Ser Arg Gly Ser Asp Gly Glu Thr Trp Trp Glu260 265 270Phe Asn Lys Phe Arg Arg Glu Met Thr Leu Thr Val Leu Asp Leu Val275 280 285Ala Leu Tyr Pro Thr His Asn Ile Lys Leu Tyr Pro Ile Pro Thr Gln290 295 300Thr Glu Leu Ser Arg Val Val Tyr Thr Asp Pro Val Gly Cys Phe Gly305 310 315 320Asn Arg Lys Ser Asp Ile Phe Ser Arg Leu Asn Phe Asp Tyr Leu Glu325 330 335Asn Arg Leu Thr Arg Pro Arg Glu Pro Phe Asn Tyr Leu Asn Ser Val340 345 350Gln Leu Phe Ala Ser Thr Val Ser Asn Ser Asn Asn Gly Glu Val Leu355 360 365Arg Gly Asn Leu Asn Lys Ile Met Phe Glu Gly Gly Trp Thr Ala Ser370 375 380Arg Ser Gly Asp Gly Val Thr Thr Gly Thr Pro Phe Ser Thr Met Asp385 390 395 400Trp Ser Tyr Gly Trp Gly Tyr Pro Arg Lys His Tyr Ala Glu Ile Thr405 410 415Ser Arg Ser Gln Ala Leu Pro Gly Leu Asn Asn Ser Ile His Val Ile420 425 430Val Gly Ile Asp Ser Phe Arg Ala Ile Gly Pro Gly Gly Gln Gly Asp435 440 445His Thr Phe Ser Leu Pro Gly Gly Asp Met Tyr Asp Cys Gly Lys Val450 455 460Gln Ile Asn Pro Leu Glu Asp Tyr Arg Asn Ser Asp His Trp Ile Ser465 470 475 480Asp Met Met Thr Ile Asn Gln Ser Val Gln Leu Ala Ser Asn Pro Thr485 490 495Gln Thr Phe Ala Phe Ser Ala Leu Ser Leu Gly Trp His His Ser Ser500 505 510Ala Gly Asn Arg Asn Val Tyr Val Tyr Asp Lys Ile Thr Gln Ile Pro515 520 525Ala Thr Lys Thr Val Arg Glu His Pro Met Ile Lys Gly Pro Gly Phe
530 535 540Thr Gly Gly Asp Leu Ala Asp Leu Ser Ser Asn Ser Asp Ile Leu Gln545 550 555 560Tyr Asp Leu Arg Ser Asp Tyr Asp Asp Arg Leu Thr Glu Asp Val Pro565 570 575Phe Arg Ile Arg Ile Arg Cys Ala Ser Ile Gly Val Ser Thr Ile Ser580 585 590Val Asp Asn Trp Gly Ser Ser Ser Pro Gln Val Thr Val Ala Ser Thr595 600 605Ala Ala Ser Leu Asp Thr Leu Lys Tyr Glu Ser Phe Gln Tyr Val Ser610 615 620Ile Pro Gly Asn Tyr Tyr Phe Asp Ser Ala Pro Arg Ile Arg Leu Leu625 630 635 640Arg Gln Pro Gly Arg Leu Leu Val Asp Arg Ile Glu Ile Ile Pro Val645 650 655Asn Phe Phe Pro Leu Ser Glu Gln Glu Asn Lys Ser Val Asp Ser Leu660 665 670Phe Ile Asn675<210>49<211>659<212>PRT<213>Bacillus thuringiensi<400>49Asn Ser Tyr Glu Asn Lys Asn Glu Tyr Glu Ile Leu Asn Asp Ser Lys1 5 10 15Lys Ser Asn Met Ser Asn Pro Tyr Leu Arg Tyr Pro Leu Ala Asn Asp20 25 30Ser Leu Ala Ser Met Gln Asn Thr Asn Tyr Lys Asp Trp Leu Thr Met35 40 45Cys Asp Arg Thr Asp Thr Asp Val Leu Ser Ser Arg Gly Ala Val Ser50 55 60Thr Gly Val Gly Met Leu Ser Thr Ile Leu Ser Leu Phe Gly Ile Pro65 70 75 80Leu Ile Gly Glu Gly Ile Asp Leu Leu Leu Gly Ala Ala Asp Phe Leu85 90 95Trp Pro Glu Ser Asp Gly Gly His Gln Tyr Thr Trp Glu Asp Leu Met100 105 110Asn His Ile Glu Glu Leu Met Asp Glu Arg Leu Glu Thr Glu Lys Arg115 120 125Thr Thr Ala Leu Asp Asp Leu Arg Gly Leu Lys Ala Leu Leu Gly Leu130 135 140Phe Arg Asp Ala Phe Asp Ser Trp Glu Lys Asn Gln Asn Asp Pro Ile145 150 155 160Ala Lys Asn Arg Val Gly Gly Tyr Phe Glu Asp Val His Thr His Phe165 170 175Val Lys Asp Met Ala Ser Ile Phe Ser Ala Thr Asn Tyr Glu Val Leu180 185 190Leu Leu Pro Val Tyr Ala Gln Ala Ala Asn Leu His Leu Leu Leu Leu195 200 205Arg Glu Gly Val Ile Tyr Gly Ser Arg Trp Gly Ile Ala Pro Ala Ala210 215 220Asp Phe Tyr His Asp Gln Leu Leu Lys Tyr Thr Ala Ile Tyr Ala Asn225 230 235 240His Cys Val Thr Trp Tyr Asn Asn Gly Leu Ala Gln Gln Lys Glu Leu245 250 255Phe Ala Lys Ser Pro Asn Trp Asn Arg Phe Asn Ala Tyr Arg Arg Asp260 265 270Met Thr Ile Thr Val Leu Asp Ile Ile Ala Leu Phe Pro Thr Tyr Asp275 280 285Ala Arg Leu Tyr Thr Lys Pro Ile Lys Thr Glu Leu Thr Arg Glu Ile290 295 300Tyr Ser Asp Val Leu Asn Leu Asp Val Tyr Gly Val Gln Gln Thr Asp305 310 315 320Leu Asn Lys Asn Glu Ala Ala Phe Thr Arg Ser Pro His Leu Val Thr
325 330 335Arg Leu Arg Gly Phe Asp Phe Tyr Thr Arg Thr Lys Tyr Ala Tyr Trp340 345 350Arg Tyr Leu Ala Gly His Thr Asn Tyr Phe Ser Phe Thr Gly Asn Gly355 360 365Thr Ile Tyr Ser Ser Ser Phe Asn Asn Trp Tyr Asp Thr Asp Met Thr370 375 380Lys Ser Thr Ile Asn Ile Pro Asp Tyr Ala Asn Ile Tyr Lys Leu Trp385 390 395 400Thr Lys Ser Tyr Thr Asn Ile Ser Pro Tyr Thr Asp Pro Val Gly Ile405 410 415Ser Gln Met Gln Phe Ser Leu Thr Asn Asn Gln Gln Leu Thr Tyr Thr420 425 430Gly Thr Ser Ala Pro Lys Tyr Pro Val Arg Glu Thr Phe Phe Glu Ile435 440 445Pro Pro Thr Asp Glu Lys Pro Leu Thr Tyr Glu Asn Tyr Ser His Ile450 455 460Leu Ser Tyr Met Thr Ser Ala Gln His Phe Gly Asp Lys Lys Ile Gly465 470 475 480Tyr Thr Phe Ala Trp Met His Glu Ser Val Asp Phe Asp Asn Arg Val485 490 495Asp Pro Asp Lys Ile Thr Gln Ile Pro Ala Val Lys Gly Asp Tyr Leu500 505 510Gln Tyr Gly Tyr Val Lys Gln Gly Pro Gly His Thr Gly Gly Asp Leu515 520 525Val Ser Met Ile Arg Thr Asp Arg Leu Gly Ile Asn Val Tyr Phe Pro530 535 540Gln Pro Leu Asp Tyr Arg Ile Arg Ile Arg Tyr Ser Thr Ser Ser Asn545 550 555 560Gly Tyr Leu Tyr Ile Tyr Ser Pro Asn Thr Lys Ile Val Tyr Leu Pro565 570 575Pro Thr Thr Leu Val Asp Gly Gln Pro Thr Phe Asp Pro Met Asp Phe580 585 590Ser Ala Phe Arg Val Val Glu Val Pro Ala Ser Phe Arg Ala Ser Val595 600 605Ala Gly Tyr Thr Asn Phe Thr Ile Glu Ala Gly Phe Gly Pro Val Tyr610 615 620Ile Asp Lys Ile Glu Phe Ile Pro Asp Asn Thr Thr Thr Leu Glu Tyr625 630 635 640Glu Gly Gly Arg Asp Leu Glu Lys Thr Lys Asn Ala Val Asn Asp Leu645 650 655Phe Thr Asn<210>50<211>558<212>PRT<213>Bacillus thuringiensi<400>50Met Phe Ile Ser Asn Ile Lys Asn Thr Leu Lys Ile Glu Thr Thr Asp1 5 10 15Tyr Glu Ile Asp Gln Ala Ala Ile Ser Ile Glu Cys Met Ser Asn Glu20 25 30His Ser Ser Lys Glu Glu Met Met Leu Trp Asp Glu Val Lys Gln Ala35 40 45Lys Gln Leu Ser Trp Ser Arg Asn Leu Leu Tyr Asn Gly Asp Phe Glu50 55 60Asp Val Ser Asn Gly Trp Lys Thr Ser Asn Thr Ile Glu Ile Arg Glu65 70 75 80Asn Ser Pro Val Phe Lys Gly His Tyr Leu His Met Phe Gly Ala Arg85 90 95Asp Ile Asp Gly Thr Leu Phe Pro Thr Tyr Ile Tyr Gln Lys Ile Glu100 105 110Glu Ser Lys Leu Lys Pro Tyr Thr Arg Tyr Arg Val Arg Gly Phe Val115 120 125Gly Ser Ser Lys Asp Leu Lys Leu Met Val Thr Arg Tyr Gly Lys Glu
130 135 140Ile Asp Ala Met Met Asn Val Pro Asn Asp Leu Ala Tyr Met Gln Pro145 150 155 160Thr Pro Ser Cys Gly Asp Ser Arg Cys Glu Ser Ser Ser Arg Tyr Val165 170 175Ser Gln Gly Tyr Pro Thr Pro Val Thr Asp Gly Tyr Ala Ser Gly Arg180 185 190Tyr Ala Cys Gln Ser Asn Arg Gly Thr Lys His Val Lys Cys His Asp195 200 205Arg His Pro Phe Asp Phe His Ile Asp Thr Gly Glu Leu Asp Thr Asn210 215 220Thr Asn Val Gly Ile Asp Val Leu Phe Lys Ile Ser Asn Pro Asp Gly225 230 235 240Tyr Ala Thr Leu Gly Asn Leu Glu Val Ile Glu Glu Gly Pro Leu Thr245 250 255Gly Glu Ala Leu Thr His Val Lys Gln Lys Glu Lys Lys Trp Lys Gln260 265 270His Met Glu Lys Lys Arg Trp Glu Thr Gln Gln Ala Tyr Asp Pro Ala275 280 285Lys Gln Ala Val Asp Ala Leu Phe Thr Asn Glu Gln Glu Leu His Tyr290 295 300His Ile Thr Leu Asp His Ile Gln Asn Ala Asp Arg Leu Ile Gln Ala305 310 315 320Ile Pro Tyr Val Tyr His Ala Trp Leu Pro Asp Ala Pro Gly Met Asn325 330 335Tyr Asp Gly Tyr Gln Gly Leu Asn Ala Arg Ile Met Gln Ala Tyr Asn340 345 350Leu Tyr Asp Ala Arg Asn Val Ile Thr Asn Gly Asp Phe Thr Gln Gly355 360 365Leu Thr Gly Trp His Ala Ala Gly Lys Ala Met Val Gln Gln Met Asp370 375 380Gly Ala Ser Val Leu Val Leu Ser Asn Trp Ser Ala Gly Val Ser Gln385 390 395 400Asn Leu His Val Gln Glu His His Gly Tyr Met Leu Arg Val Ile Ala405 410 415Lys Lys Glu Gly Pro Gly Lys Gly Tyr Val Thr Met Met Asp Cys Asn420 425 430Gly Asn Arg Glu Thr Leu Lys Phe Thr Ser Cys Glu Glu Gly Tyr Met435 440 445Thr Lys Thr Val Glu Val Phe Pro Glu Ser Asp Arg Val Arg Ile Glu450 455 460Ile Gly Glu Thr Glu Gly Thr Phe Tyr Val Asp Ser Ile Glu Leu Leu465 470 475 480Cys Met Gln Gly Tyr Ala Ser Asn Asn Asn Pro His Thr Gly Asn Met485 490 495Tyr Gly Gln Ser Tyr Asn Gly Asn Tyr Asn Gln Asn Thr Ser Asp Val500 505 510Tyr His Gln Gly Tyr Thr Asn Asn Tyr Asn Gln Asn Ser Ser Asn Met515 520 525Tyr Asn Gln Asn Tyr Thr His Asn Asp Asp Leu His Ser Gly Cys Thr530 535 540Cys Asn Gln Gly His Asn Ser Gly Cys Thr Cys Ser Gln Gly545 550 555<210>51<211>666<212>PRT<213>Bacillus thuringiensi<400>51Asn Ser Tyr Glu Asn Lys Asn Glu Tyr Glu Ile Leu Glu Ser Ser Ser1 5 10 15Asn Asn Thr Asn Met Pro Asn Arg Tyr Pro Phe Ala Asn Asp Arg Asp20 25 30Met Ser Thr Met Ser Phe Asn Asp Cys Gln Gly Ile Ser Trp Asp Glu35 40 45Ile Trp Glu Ser Ala Glu Thr Ile Thr Ser Ile Gly Ile Asp Leu Ile
50 55 60Glu Phe Leu Met Glu Pro Ser Leu Gly Gly Ile Asn Thr Leu Phe Ser65 70 75 80Ile Ile Gly Lys Leu Ile Pro Thr Asn His Gln Ser Val Ser Ala Leu85 90 95Ser Ile Cys Asp Leu Leu Ser Ile Ile Arg Lys Glu Val Ala Asp Ser100 105 110Val Leu Ser Asp Ala Ile Cys Arg Phe Leu Asp Gly Lys Leu Lys Asn115 120 125Tyr Arg Glu Tyr Tyr Leu Pro Tyr Leu Glu Ala Trp Leu Lys Asp Gly130 135 140Lys Pro Leu Gln Lys Thr Asn Asn Ser Asp Ile Gly Gln Leu Val Lys145 150 155 160Tyr Phe Glu Leu Ser Glu Arg Asp Phe Asn Glu Ile Leu Gly Gly Ser165 170 175Leu Ala Arg Asn Asn Ala Gln Ile Leu Leu Leu Pro Tyr Phe Cys Ala180 185 190Ser Cys Lys Cys Gln Leu Leu Leu Leu Arg Asp Ala Val Gln Tyr Glu195 200 205Glu Gln Trp Phe Pro Phe Leu Ser Ala Glu Asn Val Arg Ser Glu Leu210 215 220Ile Ser Pro Asn Ser Gly Cys Asp Phe Thr Gly Asp Tyr Tyr Glu Arg225 230 235 240Leu Lys Cys Lys Ile Ala Glu Tyr Thr Asp Tyr Cys Glu Tyr Trp Tyr245 250 255Gln Ala Gly Leu Asn Gln Ile Lys Gln Ala Gly Thr Gly Ala Asp Thr260 265 270Trp Ala Lys Phe Asn Lys Phe Arg Arg Glu Met Thr Leu Thr Val Leu275 280 285Asp Ile Ile Ala Ile Phe Gln Thr Tyr Asp Phe Lys Lys Tyr Pro Leu290 295 300Pro Thr His Val Glu Leu Thr Arg Glu Ile Tyr Thr Asp Pro Val Gly305 310 315 320Tyr Ser Ser Gly Thr Tyr Ser Trp Leu Lys Tyr Trp Thr Gly Ala Phe325 330 335Asn Thr Leu Glu Ala Asn Gly Thr Arg Gly Pro Gly Leu Val Thr Trp340 345 350Leu Arg Ser Ile Gly Ile Tyr Asn Glu Tyr Val Ser Arg Tyr Phe Ser355 360 365Gly Trp Val Gly Thr Arg His Tyr Glu Asp Tyr Thr Thr Gly Asn Gly370 375 380Asn Phe Gln Arg Met Ser Gly Thr Thr Ser Asn Asp Leu Arg Asp Ile385 390 395 400Ser Phe Pro Asn Ser Asp Ile Phe Lys Ile Glu Ser Lys Ala Ile Met405 410 415Asn Leu Val Gly Glu Ile Asn Ala Arg Pro Glu Tyr Arg Val Ser Arg420 425 430Ala Glu Phe Ser Glu Ser Thr Ala Phe Ile Tyr Leu Tyr Asp Ala Gly435 440 445Asn Ser Gly Leu Ser Ser Met Thr Ile Thr Ser Lys Leu Pro Gly Ile450 455 460Lys Asn Pro Glu Pro Ser Tyr Arg Asp Tyr Ser His Arg Leu Ser Asn465 470 475 480Ala Ala Cys Val Gly Ala Gly Asn Ser Arg Ile Asn Val Tyr Gly Trp485 490 495Thr His Thr Ser Met Ser Lys Tyr Asn Leu Ile Tyr Pro Asp Lys Ile500 505 510Thr Gln Ile Pro Ala Val Lys Ala Phe Asp Ile Ser Asp Thr Gly Pro515 520 525Gly Gln Val Ile Ala Gly Pro Gly His Thr Gly Gly Asn Val Val Ser530 535 540Leu Pro Tyr Tyr Ser Arg Leu Lys Ile Arg Leu Ile Pro Ala Ser Thr545 550 555 560Asn Lys Asn Tyr Leu Val Arg Val Arg Tyr Thr Ser Thr Ser Asn Gly565 570 575Arg Leu Leu Val Glu Arg Trp Ser Pro Ser Ser Ile Ile Asn Ser Tyr580 585 590Phe Phe Leu Pro Ser Thr Gly Pro Gly Asp Ser Phe Gly Tyr Val Asp
595 600 605Thr Leu Val Thr Thr Phe Asn Gln Pro Gly Val Glu Ile Ile Ile Gln610 615 620Asn Leu Asp Thr Pro Ile Asn Val Asp Lys Val Glu Phe Ile Pro Val625 630 635 640Asn Ser Thr Ala Leu Glu Tyr Glu Gly Lys Gln Ser Leu Glu Lys Ala645 650 655Gln Asp Val Val Asn Asp Leu Phe Val Lys660 665<210>52<211>558<212>PRT<213>Bacillus thuringiensi<400>52Met Phe Thr Asn Gly Thr Lys Asn Thr Leu Lys Ile Glu Thr Thr Asp1 5 10 15Tyr Glu Ile Asp Gln Ala Ala Ile Ser Ile Glu Cys Met Ser Asp Glu20 25 30His Ser Pro Lys Glu Lys Met Met Leu Trp Asp Glu Val Lys Arg Ala35 40 45Lys Leu Leu Ser Gln Ser Arg Asn Leu Leu Gln Asn Gly Asp Phe Gly50 55 60Asp Phe Tyr Gly Asn Asp Trp Lys Phe Gly Asn Asn Ile Ile Ile Gly65 70 75 80Ser Asn Asn Ser Ile Phe Lys Gly Asn Phe Leu Gln Met Ser Gly Ala85 90 95Arg Asp Ile Tyr Gly Thr Ile Phe Pro Thr Tyr Ile Tyr Gln Lys Ile100 105 110Asp Glu Ser Lys Leu Lys Pro Tyr Thr Arg Tyr Arg Val Arg Gly Phe115 120 125Val Gly Ser Ser Lys Asp Leu Arg Leu Met Val Thr Arg Tyr Gly Lys130 135 140Glu Ile Asp Ala Met Met Asn Val Pro Asn Asp Leu Ala Tyr Met Gln145 150 155 160Pro Asn Pro Ser Cys Gly Asp Ser Arg Cys Glu Ser Ser Ser Gln Tyr165 170 175Val Ser Gln Gly Tyr Pro Thr Pro Thr Asp Gly Tyr Ala Pro Asp Arg180 185 190Tyr Ala Cys Pro Ser Ser Ser Asp Lys Lys His Val Met Cys His Asp195 200 205Arg His Pro Phe Asp Phe His Ile Asp Thr Gly Glu Leu Asp Thr Asn210 215 220Thr Asn Val Gly Ile Asp Val Leu Phe Lys Ile Ser Asn Pro Asp Gly225 230 235 240Tyr Ala Thr Leu Gly Asn Leu Glu Val Ile Glu Glu Gly Pro Leu Thr245 250 255Gly Glu Ala Leu Thr His Val Lys Gln Lys Glu Lys Lys Trp Lys Gln260 265 270His Met Glu Lys Lys Arg Trp Glu Thr Gln Gln Ala Tyr Asp Pro Ala275 280 285Lys Gln Ala Val Asp Thr Leu Phe Thr Asn Glu Gln Glu Leu His Tyr290 295 300His Ile Thr Leu Asp Tyr Ile Gln Thr Leu Ile Asp Trp Tyr Ser Arg305 310 315 320Phe Pro Ile Tyr Thr Met Thr Gly Tyr Arg Asp Ala Pro Gly Met Asn325 330 335Tyr Asp Gly Tyr Gln Gly Leu Asn Ala ArgIle Met Gln A la Tyr Asn340 345 350Leu Tyr Asp Ala Arg Asn Val Ile Thr Asn Gly Asp Phe Thr Lys Gly355 360 365Leu Gln Gly Trp His Ala Ala Gly Lys Ala Ala Val Gln Gln Ile Asp370 375 380Gly Ala Ser Val Leu Val Leu Ser Asn Trp Ser Ala Gly Val Ser Gln385 390 395 400Asn Leu His Ala Gln Asp His His Gly Tyr Met Leu Arg Val Ile Ala
405 410 415Lys Lys Glu Gly Pro Gly Lys Gly Tyr Val Thr Met Met Asp Cys Asn420 425 430Gly Asn Gln Glu Thr Leu Lys Phe Thr Ser Cys Glu Glu Gly Tyr Met435 440 445Thr Lys Thr Val Glu Val Phe Pro Glu Ser Asp Arg Val Arg Ile Glu450 455 460Ile Gly Glu Thr Glu Gly Thr Phe Tyr Val Asp Ser Ile Glu Leu Leu465 470 475 480Cys Met Gln Gly Tyr Ala Ser Asn Asn Asn Pro His Thr Gly Asn Met485 490 495Tyr Gly Gln Ser Tyr Asn Gly Asn Tyr Asn Gln Asn Thr Ser Asp Val500 505 510Tyr His Gln Gly Tyr Thr Asn Asn Tyr Asn Gln Asn Ser Ser Asn Met515 520 525Tyr Asn Gln Asn Tyr Thr His Asn Asp Asp Leu His Ser Gly Cys Thr530 535 540Cys Asn Gln Gly His Asn Ser Gly Cys Thr Cys Ser Gln Gly545 550 55权利要求
1.选自下组的分离核酸分子a)含有SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26、或28所示核苷酸序列的核酸分子；b)含有与SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28所示核苷酸序列具有至少95％序列同一性的核苷酸序列的核酸分子，其中所述核苷酸序列编码具有杀虫活性的多肽；c)编码含有SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示氨基酸序列的多肽的核酸分子；d)含有编码具有与SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示氨基酸序列有至少95％氨基酸序列同一性的多肽的核苷酸序列的核酸分子，其中所述多肽具有杀虫活性；和e)a)-d)中任一项的互补物。
2.权利要求1的分离核酸分子，其中所述核苷酸序列是设计成在植物中表达的合成序列。
3.权利要求2的核酸分子，其中所述合成序列具有升高的GC含量。
4.含有权利要求1的核酸分子的载体。
5.权利要求4的载体，其中还含有编码异源多肽的核酸分子。
6.含有权利要求4的载体的宿主细胞。
7.权利要求6的宿主细胞，它是细菌宿主细胞。
8.权利要求6的宿主细胞，它是植物细胞。
9.含有权利要求8的宿主细胞的转基因植物。
10.权利要求9的转基因植物，其中所述植物选自下组玉米、高粱、小麦、向日葵、番茄、十字花科植物、胡椒属植物、马铃薯、棉花、稻、大豆、甜菜、甘蔗、烟草、大麦、和油籽油菜。
11.权利要求9的植物的转基因种子。
12.选自下组的分离多肽a)含有SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示氨基酸序列的多肽；b)由SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28所示核苷酸序列编码的多肽，其中所述多肽具有杀虫活性；c)含有与SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示氨基酸序列具有至少95％序列同一性的氨基酸序列的多肽，其中所述多肽具有杀虫活性；和d)由与SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28所示核苷酸序列至少95％相同的核苷酸序列编码的多肽。
13.权利要求12的多肽，其中还含有异源氨基酸序列。
14.选择性结合权利要求12的多肽的抗体。
15.含有权利要求12的多肽的组合物。
16.权利要求15的组合物，其中所述组合物选自下组粉剂、尘剂、丸剂、颗粒剂、喷雾剂、乳液、胶体和溶液。
17.权利要求15的组合物，其中所述组合物是通过对苏云金芽孢杆菌细胞进行干燥、冻干、匀浆、抽提、过滤、离心、沉降、或浓缩而制备的。
18.权利要求15的组合物，其中含有约1重量％至约99重量％的所述多肽。
19.用于生产具有杀虫活性的多肽的方法，其中包括在编码所述多肽的核酸分子得到表达的条件下培养权利要求6的宿主细胞，所述多肽选自下组a)含有SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示氨基酸序列的多肽；b)由SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28所示核苷酸序列编码的多肽，其中所述多肽具有杀虫活性；c)含有与SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示氨基酸序列具有至少95％序列同一性的氨基酸序列的多肽，其中所述多肽具有杀虫活性；和d)由与SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28所示核苷酸序列至少95％相同的核苷酸序列编码的多肽。
20.用于控制鳞翅类或鞘翅类害虫种群的方法，其中包括使所述种群接触杀虫有效量的权利要求12的多肽。
21.用于杀死鳞翅类或鞘翅类害虫的方法，其中包括使所述害虫接触或给其喂食杀虫有效量的权利要求12的多肽。
22.在其基因组中稳定掺入了DNA构建体的植物，其中所述DNA构建体含有编码具有杀虫活性的蛋白质的核苷酸序列，该核苷酸序列选自下组a)SEQ ID NO1、2、4、6、8、10、12、13、15、19、21、23、25、26、或28所示核苷酸序列；b)与SEQ ID NO1、2、4、6、8、10、12、13、15、19、21、23、25、26或28所示核苷酸序列具有至少95％序列同一性的核苷酸序列，其中所述核苷酸序列编码具有杀虫活性的多肽；c)编码含有SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29所示氨基酸序列的多肽的核苷酸序列；和d)编码与SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29所示氨基酸序列具有至少95％氨基酸序列同一性的多肽的核苷酸序列，其中所述多肽具有杀虫活性；其中所述核苷酸序列可操作性连接了驱动编码序列在植物细胞中表达的启动子。
23.权利要求22的植物，其中所述植物还含有SEQ ID NO17所示核苷酸序列。
24.在其基因组中稳定掺入了DNA构建体的植物细胞，其中所述DNA构建体含有编码具有杀虫活性的蛋白质的核苷酸序列，该核苷酸序列选自下组a)SEQ ID NO1、2、4、6、8、10、12、13、15、19、21、23、25、26、或28所示核苷酸序列；b)与SEQ ID NO1、2、4、6、8、10、12、13、15、19、21、23、25、26或28所示核苷酸序列具有至少95％序列同一性的核苷酸序列，其中所述核苷酸序列编码具有杀虫活性的多肽；c)编码含有SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29所示氨基酸序列的多肽的核苷酸序列；和d)编码与SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29所示氨基酸序列具有至少95％氨基酸序列同一性的多肽的核苷酸序列，其中所述多肽具有杀虫活性；其中所述核苷酸序列可操作性地连接了驱动编码序列在植物细胞中表达的启动子。
25.权利要求24的植物细胞，其中所述植物细胞还含有SEQ IDNO17所示核苷酸序列。
全文摘要
本发明提供了赋予细菌、植物、植物细胞、组织和种子以杀虫活性的组合物和方法。本发明提供了含有δ－内毒素和δ－内毒素相关多肽的编码序列的组合物。所述编码序列可用于DNA构建体或表达盒中，以便用于植物和细菌中的转化和表达。组合物还包括被转化的细菌、植物、植物细胞、组织和种子。具体而言，本发明提供了分离的δ－内毒素和δ－内毒素相关核酸分子。另外，本发明还涵盖与多核苷酸对应的氨基酸序列。具体而言，本发明提供了含有编码SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27和29中所示氨基酸序列的核苷酸序列和SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26和28中所示核苷酸序列的分离的核酸分子及其变体和片段。
文档编号C07K16/12GK1761753SQ200480007563
公开日2006年4月19日 申请日期2004年2月20日 优先权日2003年2月20日
发明者N·卡罗奇, T·哈吉斯, M·G·科奇埃尔, N·B·达克, B·卡尔 申请人:阿则耐克斯公司