δ－內毒素基因及其使用方法

2023-06-04 13:05:51

專利名稱：δ－內毒素基因及其使用方法
技術領域：
本發明涉及分子生物學領域。所提供的是編碼殺蟲蛋白質的新基因。這些蛋白質及編碼它們的核酸序列可用於製備殺蟲製劑以及生產轉基因的抗害蟲植物。
背景技術：
蘇雲金芽孢桿菌是形成孢子的革蘭氏陽性土壤細菌，特徵在於它能產生對某些目和種的昆蟲有特異性毒性、但對植物和其它非靶向生物體無害的晶狀內含物。因此，含蘇雲金芽孢桿菌菌株或它們的殺蟲蛋白質的組合物可用作環境可接受的殺蟲劑，以抑制農業有害昆蟲或針對多種人或動物疾病的昆蟲載體。
來自蘇雲金芽孢桿菌的晶體(Cry)蛋白質(δ-內毒素)具有主要針對鱗翅類、雙翅類和鞘翅類幼蟲的有效殺蟲活性。這些蛋白質還顯示出抗膜翅目、同翅目、蝨、食毛目和蟎等有害目以及諸如線蟲、扁形動物(Platyhelminthes)和Sarcomastigorphora等其它無脊椎動物目的活性(Feitelson(1993)蘇雲金芽孢桿菌家族樹，高級基因工程殺蟲劑中(The Bacillus Thuringiensis family tree.InAdvanced Engineered Pesticides).Marcel Dekker，Inc.，New York，N.Y.)。這些蛋白質主要基於其殺蟲活性而最初被劃分為CryI至CryV。主要的種類是鱗翅類特異的(I)、鱗翅類和雙翅類特異的(II)、鞘翅類特異的(III)、雙翅類特異的(IV)以及線蟲類特異的(V)和(VI)。所述蛋白質進一步被劃分為亞家族，各個家族內更密切相關的蛋白質被指派上分類字母，諸如Cry1A、Cry1B、Cry1C等。在各亞類內更緊密相關的蛋白質則命名為諸如Cry1C1、Cry1C2等。
最近出現了一種針對Cry基因的新命名法，其基於的是胺基酸序列同源性而非昆蟲靶特異性(Crickmore等(1998)Microbiol.Mol.Biol.Rev.62807-813)。在該新的分類法中，每種毒素被分配了一個獨特的名稱，其中合併了第一等級(阿拉伯數字)、第二等級(大寫字母)、第三等級(小寫字母)和第四等級(另一阿拉伯數字)。在新的分類中，羅馬數字被轉換成第一等級的阿拉伯數字。序列同一性小於45％的蛋白質具有不同的第一等級，而對於第二和第三等級而言這一標準分別為78％和95％。
晶體蛋白質在被昆蟲中腸攝取和溶解前不呈現殺蟲活性。被攝取的毒素原在昆蟲消化道內被蛋白酶水解成活性的有毒分子。(H_fte和Whiteley(1989)Microbiol.Rev.53242-255)。此毒素與靶目標幼蟲中腸內的頂端刷狀邊緣受體結合併插入頂端膜中，產生離子通道或孔，導致幼蟲死亡。
δ-內毒素通常具有5個保守的序列結構域和三個保守的結構性結構域(參閱，例如，de Maagd等(2001)Trends Genetics17193-199)。第一個保守的結構性結構域由7個α螺旋組成，並涉及膜插入和孔形成。結構域II由排列成希臘語鑰匙構形的三個β摺疊片組成，而結構域III由「果凍-卷」形式的兩個反向平行β摺疊組成(de Maagd等(2001)同上文)。結構域II和III涉及受體識別和結合，並因此被認為是毒素特異性的決定區。
因為昆蟲可造成的破壞，所以一直需要發現新形式的蘇雲金芽孢桿菌δ-內毒素。
發明簡述本發明提供了賦予細菌、植物、植物細胞、組織和種子以殺蟲劑抗性的組合物和方法。組合物包括編碼δ-內毒素和δ-內毒素相關多肽序列的分離核酸分子、包含這些核酸分子的載體和包含這些載體的宿主細胞。組合物還包括內毒素的分離或重組多肽序列、包含這些多肽的組合物以及針對那些多肽的抗體。所述核酸序列可用於DNA構建體或表達盒中，用於生物體中的轉化和表達，包括微生物和植物。所述核苷酸或胺基酸序列可以是已被設計成最適於在生物體中表達的合成序列，包括但不局限於，微生物或植物。組合物還包括被轉化的細菌、植物、植物細胞、組織和種子。
具體而言，本發明提供了含有編碼SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29中所示胺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中所給出的核苷酸序列的分離核酸分子以及它們的變體和片段。與本發明的核苷酸序列互補或與本發明的序列雜交的核苷酸序列也包括在內。
還提供了可用於產生本發明的多肽的方法，以及利用那些多肽來控制或殺死鱗翅類或鞘翅類害蟲的方法。
本發明的組合物和方法可用於產生具有殺蟲劑抗性的生物體，尤其是細菌和植物。這些生物體及其來源的組合物是農業用途所需要的。本發明的組合物還可用於產生具有殺蟲活性的改變或改良的δ-內毒素或δ-內毒素相關蛋白質，或用於檢測產品或生物體中δ-內毒素或δ-內毒素相關蛋白質或者核酸的存在。
附圖描述

圖1顯示了AXMI-004(SEQ ID NO3)與cry1Ac(SEQ ID NO31)、cry1Ca(SEQ ID NO32)、cry2Aa(SEQ ID NO34)、cry3Aa1(SEQ IDNO35)、cry1Ia(SEQ ID NO33)和cry7Aa(SEQ ID NO41)的序列比對。具有C-末端非毒性結構域的毒素如圖所示被人為截短。該序列比對顯示了以黑色標示的最高度保守胺基酸殘基和以灰色標示的高度保守胺基酸殘基。發現保守組1是從SEQ ID NO3的大約第174位至大約第196位胺基酸殘基。發現保守組2是從SEQ ID NO3的大約第250位至大約第292位胺基酸殘基。發現保守組3是從SEQ ID NO3的大約第476位至大約第521位胺基酸殘基。發現保守組4是從SEQ ID NO3的大約第542位至大約552位胺基酸殘基。發現保守組5是從SEQ IDNO3的大約第618位至大約第628位胺基酸殘基。
圖2A、B和C顯示了AXMI-006(SEQ ID NO7)與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)的序列比對。具有C-末端非毒性結構域的毒素如圖所示被人為截短。該序列比對顯示了以黑色標示的最高度保守胺基酸殘基和以灰色標示的高度保守胺基酸殘基。發現保守組1是從SEQ ID NO7的大約第218位至大約第239位胺基酸殘基。發現保守組2是從SEQ ID NO7的大約第300位至大約第350位胺基酸殘基。發現保守組3是從SEQ ID NO7的大約第547位至大約第592位胺基酸殘基。發現保守組4是從SEQ ID NO7的大約第611位至大約621位胺基酸殘基。發現保守組5是從SEQ ID NO7的大約第694位至大約第704位胺基酸殘基。
圖3A、B和C顯示了AXMI-007(SEQ ID NO9)與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)的序列比對。具有C-末端非毒性結構域的毒素如圖所示被人為截短。該序列比對顯示了以黑色標示的最高度保守胺基酸殘基和以灰色標示的高度保守胺基酸殘基。發現保守組1是從SEQ ID NO9的大約第217位至大約第238位胺基酸殘基。發現保守組2是從SEQ ID NO9的大約第299位至大約第347位胺基酸殘基。發現保守組3是從SEQ ID NO9的大約第445位至大約第590位胺基酸殘基。發現保守組4是從SEQ ID NO9的大約第609位至大約619位胺基酸殘基。發現保守組5是從SEQ ID NO9的大約第692位至大約第702位胺基酸殘基。
圖4A、B和C顯示了AXMI-008(SEQ ID NO14)與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)的序列比對。具有C-末端非毒性結構域的毒素如圖所示被人為截短。該序列比對顯示了以黑色標示的最高度保守胺基酸殘基和以灰色標示的高度保守胺基酸殘基。發現保守組1是從SEQ ID NO14的大約第185位至大約第206位胺基酸殘基。發現保守組2是從SEQ ID NO14的大約第276位至大約第318位胺基酸殘基。發現保守組3是從SEQ ID NO14的大約第497位至大約第547位胺基酸殘基。發現保守組4是從SEQ ID NO14的大約第576位至大約586位胺基酸殘基。發現保守組5是從SEQ ID NO14的大約第657位至大約第667位胺基酸殘基。
圖5A和B顯示了AXMI-008orf2(SEQ ID NO18)與cry19Aa-orf2(SEQ ID NO46)、crybun2-orf2(SEQ ID NO50)、crybun3-orf2(SEQID NO52)、cry4Aa(SEQ ID NO38)和cry4Ba(SEQ ID NO39)的序列比對。該序列比對顯示了以黑色標示的最高度保守胺基酸殘基和以灰色標示的高度保守胺基酸殘基。
圖6A、B和C顯示了AXMI-009(SEQ ID NO20)與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)的序列比對。具有C-末端非毒性結構域的毒素如圖所示被人為截短。該序列比對顯示了以黑色標示的最高度保守胺基酸殘基和以灰色標示的高度保守胺基酸殘基。發現保守組1是從SEQ ID NO20的大約第196位至大約第217位胺基酸殘基。發現保守組2是從SEQ IDNO20的大約第269位至大約第311位胺基酸殘基。發現保守組3是從SEQ ID NO20的大約第514位至大約第556位胺基酸殘基。發現保守組4是從SEQ ID NO20的大約第574位至大約584位胺基酸殘基。發現保守組5是從SEQ ID NO20的大約第651位至大約第661位胺基酸殘基。
圖7A、B和C顯示了AXMI-014(SEQ ID NO27)與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)的序列比對。具有C-末端非毒性結構域的毒素如圖所示被人為截短。該序列比對顯示了以黑色標示的最高度保守胺基酸殘基和以灰色標示的高度保守胺基酸殘基。發現保守組1是從SEQ ID NO27的大約第177位至大約第188位胺基酸殘基。發現保守組2是從SEQ ID NO27的大約第251位至大約第293位胺基酸殘基。發現保守組3是從SEQ ID NO27的大約第483位至大約第533位胺基酸殘基。發現保守組4是從SEQ ID NO27的大約第552位至大約562位胺基酸殘基。
圖8顯示了4-20％梯度SDS丙烯醯胺凝膠的照片。泳道1-4包括表達69kD AXMI-004蛋白質的各種濃度的形成孢子的芽孢桿菌細胞培養物。泳道5-8包括各種濃度的BSA。泳道9包含分子量標記物。箭頭指示69kD的帶。
發明詳述本發明針對的是用於調控生物體、尤其是植物或植物細胞中害蟲抗性的組合物和方法。所述方法包括用編碼本發明δ-內毒素或δ-內毒素相關蛋白質的核苷酸序列轉化生物體。具體而言，本發明的核苷酸序列可用於製備具有殺蟲活性的植物和微生物。因而提供了已轉化的細菌、植物、植物細胞、植物組織和種子。組合物是蘇雲金芽孢桿菌的δ-內毒素或δ-內毒素相關核酸和蛋白質。所述序列可用於構建表達載體用於隨後轉化入目的生物體內，作為探針用於分離其它的δ-內毒素或δ-內毒素相關基因，以及用本領域已知方法產生改變的殺蟲蛋白質，諸如結構域交換或DNA改組。所述蛋白質可用於抑制或殺死鱗翅類或鞘翅類有害種群以及用於產生具有殺蟲活性的組合物。
定義「δ-內毒素」指來自蘇雲金芽孢桿菌且對一種或多種害蟲具有毒性的毒素，所述害蟲包括、但不局限於鱗翅目、雙翅目和鞘翅目等的種類。在某些情況下，δ-內毒素蛋白質已從其它生物體分離出來了，包括雙酶梭狀芽孢桿菌(Clostridium bifermentans)和？類芽孢桿菌(Paenibacillus popilliae)。δ-內毒素蛋白質包含由本文所公布的全長核苷酸序列推導出來的胺基酸序列，以及由於使用備選的下遊起始位點或者由於產生具有殺蟲活性的較短蛋白質的加工所造成的比全長序列更短的胺基酸序列。加工可發生在所述蛋白質所表達的生物體內，或攝取所述蛋白質後在害蟲體內。δ-內毒素包括被確認為cry1至cry43、cyt1和cyt2的蛋白質以及Cyt-類毒素。目前有超過250種具有廣泛特異性和毒性的已知δ-內毒素。更全面的名單參閱Crickmore等(1998)，Microbiol.Mol.Biol.Rev.62807-813，而定期的更新資料參閱www.biols.susx.ac.uk/Home/Neil-Crickmore/Bt/index上的Crickmore等(2003)″蘇雲金芽孢桿菌毒素命名(Bacillus thuringiensis toxin nomenclature)″。
細菌基因，諸如本發明的AXMI基因，通常具有多個甲硫氨酸起始密碼子接近開發閱讀框架的起始點。經常地，開始於一個或多個這些起始密碼子的翻譯會導致產生功能性蛋白質。這些起始密碼子可包括ATG密碼子。不過，諸如芽孢桿菌等的細菌還將密碼子GTG識別為起始密碼子，而翻譯起始於GTG密碼子的蛋白質在第一個胺基酸處含有甲硫氨酸。此外，並非常常可以爭先確定這些密碼子中的哪一個在細菌中天然使用。因此，可以理解，利用這些變通性甲硫氨酸密碼子之一還可導致產生編碼殺蟲活性的δ-內毒素蛋白質。例如，本發明AXMI-004δ-內毒素的一個備選起始位點是SEQ ID NO1第385位鹼基對處。從此備選起始位點開始的翻譯產生了SEQ ID NO5中所顯示的胺基酸序列。本發明AXMI-007δ-內毒素蛋白質的一個備選起始位點可以是SEQ ID NO8第151位鹼基對處。從此備選起始位點開始的翻譯產生了SEQ ID NO11中所顯示的胺基酸序列。本發明的AXMI-008δ-內毒素蛋白質的一個備選起始位點可以是SEQ ID NO12的第177位核苷酸處。從此備選起始位點開始的翻譯產生了SEQ ID NO16中所示的胺基酸序列。本發明的AXMI-009δ-內毒素蛋白質的一個備選起始位點可以是SEQ ID NO19的第34位核苷酸處。從此備選起始位點開始的翻譯產生了SEQ ID NO22中所示的胺基酸序列。本發明的AXMI-009δ-內毒素蛋白質的另一個備選起始位點可以是SEQ ID NO1的第64位核苷酸處。從此備選起始位點開始的翻譯產生了SEQ IDNO24中所示的胺基酸序列。本發明的AXMI-014δ-內毒素蛋白質的一個備選起始位點可以是SEQ ID NO25的第136位核苷酸處。從此備選起始位點開始的翻譯產生了SEQ ID NO29中所示的胺基酸序列。這些δ-內毒素蛋白質都包括在本發明的範圍內，可用於本發明的方法中。
此外，在所公布的編碼一個或多個δ-內毒素相關蛋白質的核苷酸序列中可以有一個或多個另外的開發閱讀框架。「δ-內毒素相關蛋白質」指用不同於本發明δ-內毒素所用，使用另一開發閱讀框架由此處所公布的核苷酸序列編碼的蛋白質。諸如此類的蛋白質在本領域中已知可作為輔助蛋白質、穩定序列或δ-內毒素相關蛋白質。這些δ-內毒素相關蛋白質可具有殺蟲活性，或可在促進δ-內毒素蛋白質表達中起重要作用。用於檢測殺蟲活性以及用於測定δ-內毒素相關蛋白質對δ-內毒素蛋白質表達和晶體形成的影響的方法是本領域已知的(參閱，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)。這些δ-內毒素相關蛋白質包括在本發明範圍內，並可單獨或與已知的δ-內毒素蛋白質聯合用於本文所公布的方法中。在某一實施方案中，δ-內毒素相關蛋白質具有存在於SEQ ID NO18中的胺基酸序列，由SEQ ID NO17的核苷酸序列編碼。
「植物細胞」指所有已知形式的植物，包括未分化的組織(如愈傷組織)、懸浮培養細胞、原生質體、葉細胞、根細胞、韌皮部細胞、植物種子、花粉、繁殖體、胚等等。「植物表達盒」指能導致蛋白質在植物細胞中從開放閱讀框架表達的DNA構建體。一般其中包含啟動子和編碼序列。經常地，這樣的構建體還包含3』非翻譯區。所述構建體可包含「信號序列」或「前導序列」，以促進肽在翻譯的同時或翻譯後轉運至某些細胞內結構，諸如葉綠體(或其它質體)、內質網或高爾基體。
「信號序列」指已知或猜測會導致翻譯時或翻譯後的跨細胞膜肽轉運的序列。在真核細胞中，這通常包括分泌入高爾基體內，具有某些由此產生的糖基化作用。「前導序列」指在翻譯時產生的胺基酸序列足以引起肽鏈共翻譯轉運至亞細胞器的任何序列。因此，這包括通過進入內質網、穿越液泡、包括葉綠體、線粒體等在內的質體而靶向轉運和/或糖基化作用的前導序列。
「植物轉化載體」指植物細胞有效轉化所必需的DNA分子。這樣的分子可由一個或多個植物表達盒組成，且可組裝入一個以上的「載體」DNA分子中。例如，二元載體是利用兩個非毗連的DNA載體編碼植物細胞轉化所必需的所有順式和反式作用功能的植物轉化載體(Hellens和Mullineaux(2000)Trends in Plant Science5446-451)。「載體」指設計成在不同宿主細胞之間轉移的核酸構建體。「表達載體」指能在外來細胞中摻入、整合併表達異源DNA序列或片段的載體。
「轉基因植物」或「已轉化的植物」或「穩定轉化的植物或細胞組織」指在植物細胞中已摻入或整合了外源核酸序列或DNA片段的植物。這些核酸序列包括外源的或不存在於未轉化植物細胞內的那些核酸序列，以及可能是內源的或存在於未轉化的植物細胞內的那些核酸序列。「異源的」通常指對於它們所存在的細胞或部分天然基因組而言不是內源性的，且已通過感染、轉染、顯微注射、電穿孔、顯微投影(microprojection)等方式加入細胞內的核酸序列。
「啟動子」指起到指導下遊編碼序列轉錄的功能的核酸序列。啟動子與其它轉錄和翻譯調節核酸序列(也稱「調控序列」)一起是目的DNA序列表達所必需的。
本文提供了賦予殺蟲活性的新的分離核苷酸序列。還提供了δ-內毒素和δ-內毒素相關蛋白質的胺基酸序列。由此基因翻譯所產生的蛋白質可使細胞抑制或殺死攝取它的害蟲。
「分離的」或「純化的」核酸分子或蛋白質或者其生物學活性部分指的是，在用重組技術生產時基本上無其它細胞物質或培養基，或者在化學合成時基本上無化學前體或其它化學物質。優選的，「分離的」核酸中沒有天然狀態下在該核酸來源的生物體的基因組內處於該核酸兩側(即，位於所述核酸5』和3』末端的序列)的序列(優選蛋白質編碼序列)。就本發明的目的而言，「分離的」用於描述核酸分子時，並不包括分離的染色體。例如，在各種實施方案中，分離的編碼δ-內毒素或δ-內毒素相關蛋白的核酸分子可包含天然存在於所述核酸來源的細胞的基因組DNA中該核酸分子兩側的小於大約5kb、4kb、3kb、2kb、1kb、0.5kb或0.1kb的核苷酸序列。基本上無細胞物質的δ-內毒素或δ-內毒素相關蛋白質包括具有少於大約30％、20％、10％或5％(乾重)的非δ-內毒素或非δ-內毒素相關蛋白質(在此也稱為「汙染蛋白質」)的蛋白質製品。本發明的各個方面在以下部分有更進一步的詳述。
分離的核酸分子及其變體和片段本發明的一方面是關於含有編碼δ-內毒素或δ-內毒素相關蛋白質和多肽或其生物學活性部分的核苷酸序列的分離核酸分子，以及足以用作雜交探針來鑑定編碼δ-內毒素或δ-內毒素相關蛋白質的核酸分子。用於此處時，術語「核酸分子」意在包括DNA分子(如，cDNA或基因組DNA)和RNA分子(如，mRNA)和用核苷酸類似物產生的所述DNA或RNA的類似物。所述核酸分子可以是單鏈或雙鏈的，但優選是雙鏈DNA。
編碼本發明蛋白質的核苷酸序列包括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補序列。「互補序列」意指足以與給定核苷酸序列互補、從而可與該給定核苷酸序列雜交形成穩定的雙聯體的核苷酸序列。由這些核苷酸序列編碼的δ-內毒素或δ-內毒素相關蛋白質的相應胺基酸序列如SEQ ID NOS3、5、7、9、11、14、16、18、20、22、24、27和29中所示。
作為這些δ-內毒素或δ-內毒素相關蛋白質編碼核苷酸序列的片段的核酸分子也包括在本發明內。「片段」指編碼δ-內毒素或δ-內毒素相關蛋白質的核苷酸序列的一部分。核苷酸序列的片段可編碼δ-內毒素或δ-內毒素相關蛋白質的生物學活性部分，或可能是可以用下文所述方法作為雜交探針或PCR引物的片段。作為δ-內毒素或δ-內毒素相關核苷酸序列的片段的核酸分子包含至少約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個核苷酸，或者多達本文所公布的全長δ-內毒素或δ-內毒素相關蛋白質編碼核苷酸序列的核苷酸數目(例如，對於SEQ ID NO1而言是2190個核苷酸，對於SEQ ID NO2而言是1890個核苷酸，等等)，這將取決於目的用途。
本發明的核苷酸序列的片段將編碼保留δ-內毒素或δ-內毒素相關蛋白質的生物學活性、從而分別保持了殺蟲活性或δ-內毒素相關蛋白質活性的蛋白質片段。「δ-內毒素活性」指殺蟲活性。「δ-內毒素相關蛋白質活性」指所述蛋白質具有殺蟲活性，或所述蛋白質可提高δ-內毒素蛋白質的表達。這種蛋白質表達的提高可以以任何機制發生。「保持活性」指所述片段具有δ-內毒素或δ-內毒素相關蛋白質的活性的至少約30％、優選至少約50％、更優選至少約70％、還更優選至少約80％。用於測定δ-內毒素相關蛋白質對δ-內毒素蛋白質表達和晶體形成的影響的方法是本領域所已知的(參閱，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)。用於測定殺蟲活性的方法在本領域內是眾所周知的。參閱，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；和美國專利5,743,477，所有這些都全文收編於此作為參考。
編碼本發明蛋白質生物學活性部分的δ-內毒素或δ-內毒素相關蛋白質編碼核苷酸序列片段將編碼本發明全長δ-內毒素或δ-內毒素相關蛋白質中的至少約15、25、30、50、75、100、125、150、175、200、250、300、350、400、450、500、550、600或650個連續的胺基酸，或最多可達其中所存在的總胺基酸數目(例如，對於SEQ ID NO3而言是629個胺基酸，對於SEQ ID NO5而言是601個胺基酸，等等)。
優選的本發明δ-內毒素或δ-內毒素相關蛋白質是由與SEQ IDNO3、5、7、9、11、14、16、18、20、22、24、27或29中的核苷酸序列足夠一致的核苷酸序列編碼的。「足夠一致的」指用標準參數，通過本文所述序列比對程序之一與參考序列比較而言具有至少約60％或65％序列同一性、優選至少約70％或75％序列同一性、更優選至少約80％或85％序列同一性、最優選至少約90％、91％、92％、93％、94％、95％、96％、97％、98％或99％序列同一性的胺基酸或核苷酸序列。本領域技術人員應認識到，這些數值可適當調整，以確定由於密碼子簡併性、胺基酸相似性、閱讀框架定位等原因而由兩種核苷酸序列編碼的蛋白質的相應同一性。
為了確定兩種胺基酸序列或兩種核酸的同一性百分比，將這些序列進行比對以達到最佳的比較效果。兩個序列之間的同一性百分比是所述序列共有的相同位置數目的函數(即，同一性百分比＝相同位置的數目/總位置數目(如重疊位置)×100)。在某一實施方案中，所述的兩個序列長度相同。兩個序列之間的同一性百分比可以用類似於下文所述方法的技術確定，允許或不允許有缺口。在計算同一性百分比時，通常嚴格匹配的才被計算在內。
兩個序列之間同一性百分比的確定可用數學運算法則完成。用於比較兩個序列的數學運算法則的非局限性例子是文獻Karlin和Altschul(1990)Proc.Natl.Acad.Sci.USA 872264中提出的運算法則，根據文獻Karlin和Altschul(1993)Proc.Natl.Acad.Sci.USA 905873-5877進行修正。這樣的運算法則被合併入文獻Altschul等(1990)J.Mol.Biol.215403的BLASTN和BLASTX程序中。BLAST核苷酸搜索可用BLASTN軟體進行(得分＝100，詞長＝12)，以獲得與本發明δ-內毒素或δ-內毒素相關核酸分子同源的核苷酸序列。BLAST蛋白質搜索可用BLASTN軟體進行(得分＝50，詞長＝3)，以獲得與本發明δ-內毒素或δ-內毒素相關蛋白質分子同源的胺基酸序列。為了獲得帶缺口的序列比對以進行比較，可如文獻Altschul等(1997)Nucleic Acids Res.253389中所述利用Gapped BLAST。或者，可用PSI-Blast進行檢測兩個分子之間疏遠關聯的反覆搜索。參閱，Altschul等(1997)，同上文。在應用BLAST、Gapped BLAST和PSI-Blast程序時，可使用各個程序(如，BLASTX和BLASTN)默認的參數。見www.ncbi.nlm.nih.gov。另一用於比較序列的數學運算法則的非限制性例子是ClustalW運算法則(Higgins等(1994)Nucleic AcidsRes.224673-4680)。ClustalW可比較序列，並將胺基酸或DNA序列全部比對，因此可提供有關整個胺基酸序列的序列保守性的資料。ClustalW運算法則被用於數個商品化的DNA/胺基酸分析軟體包中，諸如載體NTi Program Suite(Informax，Inc)的ALIGNX模塊。用ClustalW進行胺基酸序列的比對後，可評估胺基酸同一性百分比。可用於ClustalW序列比對分析的軟體程序的非局限性例子是GeneDocTM。GenedocTM(Karl Nicholas)可評估多個蛋白質之間的胺基酸(或DNA)相似性和同一性。用於序列比較的數學運算法則的另一非限制性例子是文獻Myers和Miller(1988)CABIOS 411-17中的運算法則。這樣的運算法則被引入ALIGN程序(版本2.0)中，它是GCG序列比對軟體包的一部分(可獲自Accelrys，Inc.，9865Scranton Rd.，San Diego，California，USA)。當應用ALIGN程序來比較胺基酸序列時，可使用PAM120加權殘基表、缺口長度罰分12和缺口罰分4。
本發明還包括變體核酸分子。δ-內毒素或δ-內毒素相關蛋白質編碼核苷酸序列的「變體」包括編碼本文所公布的δ-內毒素或δ-內毒素相關蛋白質、但由於遺傳密碼簡併性而有保守性差別的序列，以及如上所述足夠相似的那些序列。天然存在的等位基因變體可用眾所周知的分子生物學技術鑑定，諸如聚合酶鏈式反應(PCR)和下文概述的雜交技術。變體核苷酸序列還包括合成來源的核苷酸序列，例如如下所述通過定位誘變產生、但仍然編碼本發明公布的δ-內毒素或δ-內毒素相關蛋白質的序列。包含在本發明範圍內的變體蛋白質是有生物學活性的，也就是說，它們繼續具有天然蛋白質的目的生物學活性，即保持殺蟲活性。「保持活性」是指變體具有δ-內毒素或δ-內毒素相關蛋白質的活性的至少約30％、優選至少約50％、更優選至少約70％、還更優選至少約80％。用於測量殺蟲活性的方法在本領域中是眾所周知的。參閱，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；以及美國專利5,743,477，所有這些均全部收編於此作為參考。
本發明還包括變體核酸分子。δ-內毒素或δ-內毒素相關編碼核苷酸序列的「變體」包括那些編碼本文所公布的δ-內毒素或δ-內毒素相關蛋白質、但由於遺傳密碼簡併性而有保守性差異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變體可用眾所周知的分子生物學技術鑑定，諸如聚合酶鏈式反應(PCR)和下述雜交技術。變體核苷酸序列還包括合成來源的核苷酸序列，例如如下所述通過定位誘變產生、但仍然編碼本發明公布的δ-內毒素或δ-內毒素相關蛋白質的序列。
技術熟練人員應認識到，可通過突變在本發明的核苷酸序列中引入變異，從而導致所編碼的δ-內毒素或δ-內毒素相關蛋白質的胺基酸序列中的變化，但不改變所述蛋白質的生物學活性。因此，可通過在本文所公布的相應核苷酸序列中引入一個或多個核苷酸取代、添加或缺失而製備變異的分離核酸分子，從而在所編碼的蛋白質中引入一個或多個胺基酸取代、添加或缺失。突變可通過標準技術引入，諸如定位誘變和PCR介導誘變。這樣的變異核苷酸序列也包括在本發明的範圍內。
例如，優選的，可以在一個或多個預知的、優選非必需胺基酸殘基處進行保守胺基酸取代。「非必需的」胺基酸殘基是可以在δ-內毒素或δ-內毒素相關蛋白質序列的野生型序列中發生改變、而不改變生物學活性的殘基，而「必需的」胺基酸殘基是生物學活性所必需的。「保守的胺基酸取代」是用具有相似側鏈的胺基酸殘基替換某一胺基酸殘基。具有相似側鏈的胺基酸殘基家族在本領域中已有定義。這些家族包括具有鹼性側鏈的胺基酸(如，賴氨酸、精氨酸、組氨酸)、具有酸性側鏈的胺基酸(如，天冬氨酸、穀氨酸)、具有不帶電荷的極性側鏈的胺基酸(如，甘氨酸、天冬醯胺、穀氨醯胺、絲氨酸、蘇氨酸、酪氨酸、半胱氨酸)、具有非極性側鏈的胺基酸(如，丙氨酸、纈氨酸、亮氨酸、異亮氨酸、脯氨酸、苯丙氨酸、甲硫氨酸、色氨酸)、具有β-分支側鏈的胺基酸(如，蘇氨酸、纈氨酸、異亮氨酸)和具有芳香族側鏈的胺基酸(如，酪氨酸、苯丙氨酸、色氨酸、組氨酸)。
在δ-內毒素蛋白質之間通常有5個高度保守的區域，主要集中在結構域的中心或結構域之間的連接處(Rajamohan等(1998)Prog.Nucleic Acid Res.Mol.Biol.601-23)。多種δ-內毒素的保守胺基酸塊以及共有序列可參閱以下文獻Schnepf等(1998)Microbio.Mol.Biol.Rev.62775-806和Lereclus等(1989)，編碼蘇雲金芽孢桿菌類芽孢δ-內毒素的基因的作用、結構和分子組成(Role，Structure，and Molecular Organization of the Genes Coding forthe Parasporal d-endotoxins of Bacillus thuringiensis)。在「原核發育的調控(Regulation of Procaryotic Development)」一書中，Issar Smit，Slepecky，R.A.，Setlow，P.美國微生物學會，Washington，D.C.20006。已提出具有這些保守區的δ-內毒素可共享相似的結構，由三個結構域組成(Li等(1991)Nature 353815-821)。結構域I在δ-內毒素之間具有最高的相似性(Bravo(1997)J.Bacteriol.1792793-2801)。
可在非保守區內進行保持活性的胺基酸取代。一般而言，不對保守胺基酸殘基、或定位在保守基元內的胺基酸殘基進行這樣的取代，其中所述的殘基對於蛋白質活性是必需的。保守的、並且可能為蛋白質活性所必需的殘基例子包括，例如，在所提供的序列比對中含有的所有蛋白質之間均相同的殘基。保守的、但可能允許發生保守性胺基酸取代而仍保持活性的殘基的例子包括例如在所提供的序列比對中所包括的所有蛋白質之間僅有保守性取代的殘基。不過，本領域技術人員應理解，功能性變體可在保守殘基中具有微小的保守或非保守變異。
或者，可通過諸如飽和誘變等方法沿著所有或部分編碼序列隨機引入突變而製備變異核苷酸序列，並可對所產生的突變體篩選賦予δ-內毒素或δ-內毒素相關活性的能力，以鑑別保持活性的突變體。誘變後，所編碼的蛋白質可重組表達，而蛋白質的活性可用標準檢測技術測定。
用諸如PCR、雜交等方法可鑑定相應的δ-內毒素或δ-內毒素相關序列，這樣的序列與本發明的序列基本上相同。參閱，例如，SambrookJ.和Russell，D.W.(2001)分子克隆實驗室手冊MolecularCloningA Laboratory Manual.(冷泉港實驗室出版社，冷泉港，紐約)和Innis等(1990)PCR流程方法和應用指南PCR ProtocolsA Guide to Methods and Applications(Academic Press，NY)。
在雜交方法中，可用全部或部分δ-內毒素或δ-內毒素相關核苷酸序列篩選cDNA或基因組文庫。所述cDNA和基因組文庫的構建方法是本領域普遍已知的，並被描述在Sambrook和Russell，2001中。所謂的雜交探針可以是基因組DNA片段、cDNA片段、RNA片段或其它寡核苷酸，它們可用諸如32P等可檢測基團或諸如其它放射性同位素、螢光化合物、酶或酶輔因子等任何其它可檢測標記物進行標記。可通過標記基於本文所公布的已知δ-內毒素或δ-內毒素相關編碼核苷酸序列的合成寡核苷酸製備用於雜交的探針。此外，還可使用基於核苷酸序列或所編碼的胺基酸序列中的保守核苷酸或胺基酸殘基設計的簡併引物。所述探針通常包含在嚴謹條件下與本發明δ-內毒素或δ-內毒素相關編碼核苷酸序列或其片段或變體的至少約12個、優選約25個、更優選至少約50、75、100、125、150、175、200、250、300、350或400個連續核苷酸雜交的核苷酸序列區。用於雜交的探針的製備是本領域所普遍已知的，並且公開在此處收編作為參考的文獻Sambrook和Russell，2001中。
在雜交技術中，已知的核苷酸序列的全部或部分被用作與來自選定生物體的克隆基因組DNA片段或cDNA片段群(即，基因組或cDNA文庫)中存在的其它相應核苷酸序列選擇性雜交的探針。雜交探針可以是基因組DNA片段、cDNA片段、RNA片段或其它寡核苷酸，且可用諸如32P等可檢測基團或任何其它可檢測標記物標記。因此，例如，用於雜交的探針可通過標記基於本發明的δ-內毒素或δ-內毒素相關序列的合成寡核苷酸而製備。雜交探針的製備方法以及cDNA和基因組文庫的構建方法是本領域通常已知的，並公開在Sambrook等(1989)分子克隆實驗室手冊Molecular CloningA Laboratory Manual(第二版，冷泉港實驗室出版社，Plainview，紐約)中。
例如，本文所公布的整個δ-內毒素或δ-內毒素相關序列或其一個或多個部分可用作能特異性地與相應δ-內毒素或δ-內毒素相關樣序列以及信使RNA雜交的探針。為了實現各種條件下的特異雜交，所述探針包含獨特的序列，並且優選長度為至少約10個核苷酸、最優選至少約20個核苷酸。這樣的探針可用於通過PCR從選定的生物體中擴增相應的δ-內毒素或δ-內毒素相關序列。這一技術可用於從目的生物體中分離額外的編碼序列，或作為診斷檢測法用於確定生物體中編碼序列的存在與否。雜交技術包括鋪板的DNA文庫(或者噬斑或者菌落，參閱，例如，Sambrook等(1989)分子克隆實驗室手冊(第二版，冷泉港實驗室出版社，Plainview，紐約))的雜交篩選。
這些序列的雜交可在嚴謹條件下進行。「嚴謹條件」或「嚴謹的雜交條件」指，在此條件下，探針與其靶序列的雜交程度可檢測性地高於其它序列(例如，至少高於背景2倍)。嚴謹條件是隨序列而定的，在不同的環境下有所不同。通過控制雜交和/或漂洗條件的嚴謹性，可鑑別與探針100％互補的靶序列(同源探測)。或者，可將嚴謹條件調整成允許序列中有一些錯配，從而檢測較低程度的相似性(異源探測)。通常，探針的長度小於約1000個核苷酸，優選長度小於500個核苷酸。
通常，嚴謹條件是這樣的條件，其中在pH7.0至8.3時鹽濃度小於約1.5M Na離子、通常約0.01至1.0M Na離子濃度(或其它鹽)，並且對於短探針(如10-50個核苷酸)而言溫度至少約30℃，而對於長探針(例如，大於50個核苷酸)而言至少約60℃。還可通過添加諸如甲醯胺等去穩定劑來達到嚴謹條件。示範性的低嚴謹條件包括用30-35％甲醯胺、1M NaCl、1％SDS(十二烷基硫酸鈉)的緩衝液在37℃進行雜交，在1X至2X SSC(20X SSC＝3.0M NaCl/0.3M檸檬酸三鈉)在50-55℃漂洗。示範性的中等嚴謹條件包括在40-45％甲醯胺、1.0M NaCl、1％SDS中於37℃雜交，並在0.5X至1X SSC於55-60℃漂洗。示範性的高嚴謹條件包括在50％甲醯胺、1M NaCl、1％SDS中於37℃進行雜交，並在0.1X SSC中於60-65℃漂洗。任選的，漂洗緩衝液中可包含約0.1％至約1％SDS。雜交的持續時間一般少於大約24小時，通常約為4至12小時。
一般而言，特異性是雜交後漂洗條件的函數，關鍵因素是最終漂洗溶液的離子強度和溫度。對於DNA-DNA雜交，Tm可用Meinkoth等Wahl(1984)Anal.Biochem.138267-284中的公式估計Tm＝81.5℃+16.6(log M)+0.41(％GC)-0.61(％form)-500/L；其中的M是單價陽離子的摩爾濃度，％GC是DNA中鳥嘌呤和胞嘧啶核苷酸的百分比，％form是雜交溶液中甲醯胺的百分比，而L是鹼基對中雜交體的長度。Tm是互補靶序列的50％與完全匹配的探針雜交的溫度(在確定的離子強度和pH下)。對於每1％的錯配，Tm下降約1℃；因此，為了與所期望的同一性的序列雜交，可以調節Tm、雜交和/或漂洗條件。例如，如果搜尋具有≥90％同一性的序列，Tm可下降10℃。通常，將嚴謹條件選擇成比該具體序列與其互補序列在確定的離子強度和pH的熱融點(Tm)低約5℃。不過，嚴格的嚴謹條件可利用比熱融點(Tm)低1、2、3或4℃的條件進行雜交和/或漂洗；中等的嚴謹條件可利用比熱融點(Tm)低6、7、8、9或10℃的條件進行雜交和/或漂洗；低嚴謹條件可利用可利用比熱融點(Tm)低11、12、13、14、15或20℃的條件進行雜交和/或漂洗。利用所述公式、雜交和漂洗組合物以及預期的Tm，普通技術人員可理解雜交和/或漂洗溶液的嚴謹性的變化已得到了本質的描述。如果所需程度的錯配導致Tm小於45℃(水溶液)或32℃(甲醯胺溶液)，則優選提高SSC濃度，以利用較高的溫度。有關核酸雜交的詳盡指南可參閱文獻Tijssen(1993)生化和分子生物學實驗技術-用核酸探針進行雜交(Laboratory Techniques inBiochemistry and Molecular Biology-Hybridization with NucleicAcid Probes)，第I部分，第二章(Elsevier，紐約)；和Ausubel等編輯(1995)分子生物學通用方法(Current Protocols inMolecular Biology)，第二章(Greene Publishing andWiley-Interscience，New York)。參閱Sambrook等(1989)分子克隆實驗室手冊(Molecular CloningA Laboratory Manual)(第二版，冷泉港實驗室出版社，Plainview，New York)。
分離的蛋白質及其變體和片段
δ-內毒素和δ-內毒素相關蛋白質也包含在本發明的範圍內。「δ-內毒素蛋白質」指具有SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29中所示胺基酸序列的蛋白質。「δ-內毒素相關蛋白質」指具有SEQ ID NO18中所示胺基酸序列的蛋白質。同時還提供了它們的片段、生物學活性部分和變體，它們可用於實踐本發明的方法。「片段」或「生物學活性部分」包括含有編碼如SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29中所示δ-內毒素或δ-內毒素相關蛋白質的胺基酸序列的一部分且保持δ-內毒素活性或δ-內毒素相關活性的多肽片段。δ-內毒素或δ-內毒素相關蛋白質的生物學活性部分可以是例如10、25、50、100或更多個胺基酸長的多肽。這樣的生物學活性部分可通過重組技術製備，並評估其δ-內毒素或δ-內毒素相關活性。檢測殺蟲活性的方法是本領域眾所周知的。參閱，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；以及美國專利5,743,477，所有這些均全部收編於此作為參考。用於此處時，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個毗連胺基酸。不過，本發明還包含其它的片段，諸如該蛋白質中的任何大於約10、20、30、50、100、150、200、250、300、350、400、450、500、550、600和650個胺基酸的片段。
「變體」指具有與SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示胺基酸序列至少約60％、65％、優選至少約70％、75％、更優選至少約80％、85％、最優選至少約90％、91％、92％、93％、94％、95％、96％、97％、98％或99％相同的胺基酸序列的蛋白質或多肽。變體還包括在嚴謹條件下與SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28的核酸分子或其互補分子雜交的核酸分子編碼的多肽。這樣的變體通常保持δ-內毒素或δ-內毒素相關活性。變體包括由於誘變而胺基酸序列有所區別的多肽。本發明所包含的變體蛋白質是有生物學活性的，即，它們繼續具有天然蛋白質的所需生物學活性，即保持殺蟲活性。檢測殺蟲活性的方法是本領域眾所周知的。參閱，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；以及美國專利5,743,477，所有這些均全部收編於此作為參考。
改變或改良的變體已認識到可用各種方法改變δ-內毒素或δ-內毒素相關蛋白質的DNA序列，並且這些改變可能產生所編碼的蛋白質胺基酸序列不同於本發明δ-內毒素或δ-內毒素相關蛋白質胺基酸序列的DNA序列。這一蛋白質可以多種方式改變，包括胺基酸取代、缺失、截短和插入。這些操作方法是本領域通常已知的。例如，δ-內毒素或δ-內毒素相關蛋白質的胺基酸序列變體可通過DNA中的突變來製備。這還可以通過數種誘變形式之一和/或定向進化來完成。在某些方面，胺基酸序列中編碼的改變基本上不會影響蛋白質的功能。這樣的變體將具有所需的殺蟲活性。不過，應理解δ-內毒素或δ-內毒素相關蛋白質賦予殺蟲活性的能力可基於本發明組合物通過使用所述技術得以提高。例如，可以在於DNA複製期間呈現出高比率鹼基錯誤摻入的宿主細胞中表達δ-內毒素或δ-內毒素相關蛋白質，諸如XL-1 Red(Stratagene)。在所述株系中增殖後，可分離δ-內毒素或δ-內毒素相關DNA(例如通過製備質粒，或通過PCR擴增並將產生的PCR片段克隆入載體中)，在非誘變性菌株中培養δ-內毒素或δ-內毒素相關突變，鑑定具有殺蟲活性的突變的δ-內毒素或δ-內毒素相關基因，例如進行檢測來測試殺蟲活性。通常，將所述蛋白質混和並用於餵服試驗中。參閱，例如，Marrone等(1985)J.of Economic Entomology78290-293。這樣的試驗可包括使植物接觸一種或多種害蟲並測定植物存活的能力和/或引起害蟲死亡的能力。導致毒性增強的突變的例子參閱Schnepf等(1998)Microbiol.Mol.Biol.Rev.62775-806。
或者，可在氨基或羧基末端對許多蛋白質的序列進行改變而基本上不影響其活性。這可包括利用現代分子方法引入的插入、刪除或改變，諸如PCR，包括由於在PCR擴增所用的寡核苷酸中含有胺基酸編碼序列而改變或延長蛋白質編碼序列的PCR擴增。或者，所加入的蛋白質序列可包括完整的蛋白質編碼序列，諸如那些在本領域中通常用於產生蛋白質融合體的序列。這樣的融合蛋白質常常用於(1)增加目的蛋白質的表達，(2)引入結合結構域、酶促活性或表位以方便蛋白質純化、蛋白質檢測或本領域已知的其它實驗性應用，(3)使蛋白質的分泌或翻譯定向於亞細胞性細胞器，諸如革蘭氏陰性細菌的周質空間，或真核細胞的內質網，後者常常導致蛋白質的糖基化。
本發明的變體核苷酸和胺基酸序列還包括來自諸如DNA改組等誘變和重組方法的序列。通過這樣的步驟，可利用一個或多個不同的δ-內毒素或δ-內毒素相關蛋白質編碼區構建具有所需特性的新的δ-內毒素或δ-內毒素相關蛋白質。以此方式，由含有具有充分的序列同一性且可體外或體內同源重組的序列區的相關序列多核苷酸群產生重組多核苷酸文庫。例如，用此方法，可在本發明的δ-內毒素或δ-內毒素相關基因與其它已知δ-內毒素或δ-內毒素相關基因之間對編碼目的結構域的序列基元進行改組，以獲得編碼具有改良目的特性的蛋白質的新基因，諸如提高殺蟲活性。所述的DNA改組策略是本領域所已知的。參閱，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；以及美國專利5,605,793和5,837,458。
結構域交換或改組是產生改變的δ-內毒素或δ-內毒素相關蛋白質的另一機制。結構域II和III可在δ-內毒素蛋白質之間進行交換，產生具有改良的殺蟲活性或目標譜的雜合或嵌合毒素。產生重組蛋白質並檢測其殺蟲活性的方法在本領域中是眾所周知的(參閱，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)。
植物轉化可通過本領域已知的數種技術之一完成植物細胞的轉化。首先，以允許其在植物細胞中表達的方式改造δ-內毒素或δ-內毒素相關基因。通常表達這種蛋白質的構建體應含有驅使基因轉錄的啟動子以及允許轉錄終止和聚腺苷酸化的3』非翻譯區。這樣的構建體的組成在本領域中是眾所周知的。在某些情況下，可有效改造基因，使得產生的肽被分泌出來，或定向於植物細胞中。例如，可改造基因，使其含有信號肽，以方便所述的肽轉移至內質網。還可能優選的是，改造植物表達盒使其含有內含子，從而使內含子的mRNA加工為表達所必需。
通常這一「植物表達盒」將被插入「植物轉化載體」中。該植物轉化載體可由完成植物轉化所需的一個或多個DNA載體組成。例如，本領域的通用慣例是利用由一個以上毗連DNA區段組成的植物轉化載體。這些載體在本領域中常常稱為「二元載體」。二元載體以及具有輔助質粒的載體是最常用於農桿菌介導的轉化中的，其中完成有效轉化所需的DNA區段的大小和複雜性是相當大的，並且將功能分攤到分開的DNA分子上比較有利。二元載體通常包含含有T-DNA轉移所必需的順式作用序列(諸如左邊界和右邊界)、經改造能在植物細胞中表達的可選擇標記物以及「目的基因」(經改造能在期望產生轉基因植物的植物細胞中表達的基因)的質粒載體。還存在於此質粒載體中的是細菌複製所必需的序列。順式作用序列以可有效轉移入植物細胞並在其中表達的形式安排。例如，可選擇標記基因和目的基因定位於左和右邊界之間。常常有另一個質粒載體包含介導T-DNA從農桿菌轉移至植物細胞的的反式作用因子。按照本領域中所理解的，此質粒常常具有使植物細胞可被農桿菌感染並通過在邊界序列處的斷裂以及病毒介導的DNA轉移來轉移DNA的致病作用(Vir基因)(Hellens和Mullineaux(2000)Trends in Plant Science，5446-451)。數種類型的農桿菌菌株(如LBA4404、GV3101、EHA101、EHA105等)可用於植物轉化。該第二種質粒載體不是用諸如顯微投影、顯微注射、電穿孔、聚乙二醇等其它方法轉化植物所必需的。
一般而言，植物轉化方法包括將異源DNA轉移入靶植物細胞(如，不成熟或成熟的胚、懸浮培養物、未分化的愈傷組織、原生質體，等等)中，隨後進行最大極限水平的適當選擇(取決於可選擇標記基因)，以從未轉化細胞集合組中回收轉化的植物細胞。通常將外植體轉移至新鮮置換的同種培養基中並進行常規培養。隨後，在放置於補充了最大極限水平的選擇劑的再生培養基中後已轉化的細胞被分化成芽。然後將所述的芽轉移到選擇性生根培養基中以回收生根的芽或苗。然後將轉基因苗培養成成熟的植物並產生能繁殖的種子(如Hiei等(1994)The Plant Journal 6271-282；Ishida等(1996)NatureBiotechnology 14745-750)。通常將外植體轉移至新鮮補充的同種培養基中並進行常規培養。產生轉基因幼苗的有關技術和方法的全面描述參閱文獻Ayres和Park，1994(Critical Reviews in PlantScience 13219-239)以及Bommineni和Jauhar，1997(Maydica 42107-120)。由於被轉化的物質包含許多細胞，轉化和未轉化的細胞都存在於任何受試靶愈傷片或組織或細胞組中。殺死未轉化細胞並允許轉化細胞增殖的能力使被轉化的植物培養物得以生長。通常，去除未轉化細胞的能力是對快速回收被轉化植物細胞並成功產生轉基因植物的限制。
轉基因植物的產生可用數種方法之一完成，包括但不局限於通過農桿菌將異源DNA引入植物細胞中(農桿菌介導的轉化)、用粘附於顆粒上的外源性外來DNA轟擊植物細胞以及各種其它非粒子定向介導的方法(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)轉移DNA。
轉化方法以及將核苷酸序列引入植物的方法可能隨著待轉化的植物或植物細胞的類型(即單子葉或雙子葉植物)而變化。將核苷酸序列引入植物細胞並隨後插入植物基因組中的合適方法包括顯微注射(Crossway等(1986)Biotechniques 4320-334)、電穿孔(Riggs等(1986)Proc.Natl.Acad.Sci.USA 835602-5606)、農桿菌介導的轉化(美國專利5,563,055；美國專利5,981,840)、直接基因轉移(Paszkowski等(1984)EMBO J.32717-2722)和彈道顆粒加速(參閱，例如，美國專利4,945,050；美國專利5,879,918；美國專利5,886,244；美國專利5,932,782；Tomes等(1995)″通過顯微投射轟擊將DNA定向轉移入完整的植物細胞中(Direct DNA Transfer into Intact PlantCells via Microprojectile Bombardment)″，在「植物細胞、組織和器官培養基本方法(Plant Cell，Tissue，and Organ CultureFundamental Methods)」一書中，Gamborg和Phillips編輯(Springer-Verlag，Berlin)；McCabe等(1988)Biotechnology6923-926)、氣溶膠束轉化(美國已出版專利申請20010026941；美國專利4,945,050；國際專利申請WO91/00915；美國已出版專利申請2002015066)以及Lec1轉化(WO00/28058)。也可參閱Weissinger等(1988)Ann.Rev.Genet.22421-477；Sanford等(1987)微粒科學和技術(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；美國專利5,240,855；美國專利5,322,783和5,324,646；Tomes等(1995)″通過微粒轟擊將DNA定向轉移入完整的植物細胞中(Direct DNATransfer into Intact Plant Cells via MicroprojectileBombardment)″，在「植物細胞、組織和器官培養基本方法(PlantCell，Tissue，and Organ CultureFundamental Methods)」一書中，編輯Gamborg和Phillips(Springer-Verlag，Berlin)；Klein等(1988)Plant Physiol.91440-444；Hooykaas-Van Slogteren等(1984)Nature(London)311763-764；美國專利5,736,369；Bytebier等(1987)Proc.Natl.Acad.Sci.USA845345-5349(Liliaceae)；De Wet等(1985)在「胚珠組織的實驗操作(The ExperimentalManipulation of Ovule Tissues)」一書中，編輯Chapman等(Longman，New York)，第197-209頁；Kaeppler等(1990)植物細胞報告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，所有這些均收編於此作為參考。
在將外源性外來DNA整合入植物細胞中後，在培養基中進行最大極限水平的適當選擇，以殺死未轉化的細胞，並通過定期轉移至新鮮培養基中而分離和增殖在此選擇處理中存活的據推定已轉化的細胞。通過連續傳代並進行適當的選擇，可識別並增殖被所述質粒載體轉化的細胞。然後用分子和生化方法證實轉基因植物基因組中被整合的異源目的基因的存在。
已被轉化的細胞可以傳統方式生長成植物。參閱，例如McCormick等(1986)Plant Cell Reports 581-84。然後可培養這些植物，用同樣的被轉化株系或不同的株系授粉，並鑑定具有所需表型特徵的組成型表達的所得雜合體。可培養兩代或多代，以保證所需表型特徵的表達穩定維持和遺傳，然後收穫種子以保證已實現了所需表型特徵的表達。以此方式，本發明提供了在其基因組中穩定摻入了本發明的核苷酸構建體、例如本發明的表達盒的已轉化種子(也稱為「轉基因種子」)。
本發明的δ-內毒素或δ-內毒素相關序列可存在於用於在目的植物中表達的表達盒中。所述的盒將包括可操作性的與本發明序列連接的5′和3′調控序列。「可操作性的連接」指在啟動子和第二序列之間的功能性聯接，其中的啟動子序列起始並介導了與第二序列相應的DNA序列的轉錄。通常，可操作性連接意味著被連接的核酸序列是毗鄰的，並且，在需連接兩個蛋白質編碼區時，則是毗鄰且處於同一閱讀框架中。所述的盒可另外含有至少一個其它基因以共轉化入生物體中。或者，所述的其它基因可存在於多個表達盒中。
向所述的表達盒中提供多個限制性位點，以供插入δ-內毒素或δ-內毒素相關序列，處於調控區的轉錄調控下。
所述的表達盒沿5′-3′轉錄方向將包含在植物中發揮作用的轉錄和翻譯起始區(即，啟動子)、本發明的DNA序列以及轉錄和翻譯終止區(即終止區)。啟動子對於植物宿主和/或本發明的DNA序列而言可能是天然的或類似的、或者外源的或異源的。此外，啟動子可以是天然序列或可以是合成序列。啟動子對於植物宿主而言是「天然的」或「同源的」，指的是啟動子在該啟動子將被引入的天然植物中存在。啟動子對於本發明DNA序列而言是「外源的」或「異源的」，指的是啟動子對於可操作性連接的本發明DNA序列而言不是天然或自然存在的啟動子。
終止區可以是對於轉錄起始區而言是天然的，可以是就可操作性連接的目的DNA序列而言是天然的，可以是就植物宿主而言是天然的，或者可以來自其它來源(即，對於啟動子、目的DNA序列、植物宿主或它們的任何組合而言是外源的或異源的)。方便的終止區可從根癌農桿菌的Ti-質粒獲得，諸如章魚鹼合酶和胭脂鹼合酶終止區。還可參閱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。
在適當的情況下，可將基因最優化以提高其在被轉化宿主細胞中的表達。即，所述基因可以用宿主細胞偏好的密碼子合成以促進表達，或者可以宿主偏好的密碼子使用頻率用密碼子合成。例如，有關宿主偏好的密碼子使用的討論參閱Campbell和Gowri(1990)PlantPhysiol.921-11。合成植物偏好基因的方法是本領域所已知的。參閱，例如，美國專利6,320,100、6,075,185、5,380,831和5,436,391，美國已出版專利申請20040005600和20010003849，以及Murray等(1989)Nucleic Acids Res.17477-498，在此收編作為參考。
在一個實施方案中，目的核酸被定向於葉綠體中表達。以此方式，當目的核酸不是直接插入葉綠體的情況下，表達盒將另外含有編碼轉運肽的核酸以指導目的基因產物進入葉綠體。這樣的轉運肽在本領域中是已知的。參閱，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。
用於葉綠體轉化的方法在本領域中是已知的。參閱，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。所述方法依賴於含可選擇標記的DNA的粒子槍投遞以及通過同源重組將DNA定向於質體基因組。此外，質體轉化可通過用核編碼及質體指導的RNA聚合酶的組織優選表達而反式激活沉默的質體攜帶的轉基因而完成。這樣的體系已被報導於文獻McBride等(1994)Proc.Natl.Acad.Sci.USA 917301-7305中。
可優化定向於葉綠體的目的核酸以表達於葉綠體中，從而解決植物細胞核和此細胞器之間的密碼子使用中的差異。以此方式，目的核酸可以用葉綠體偏好的密碼子合成。參閱，例如，美國專利5,380,831，在此收編作為參考。
植物轉化的評估將異源性的外來DNA引入植物細胞中後，用各種方法證實植物基因組中異源基因的轉化或整合，諸如與被整合基因相關的核酸、蛋白質和代謝物的分析。
PCR分析PCR分析是在移植至土壤中之前的較早階段篩選已轉化的細胞、組織或芽是否存在被摻入基因的快速方法(Sambrook和Russell，2001)。PCR是用特異於目的基因或農桿菌載體背景等的寡核苷酸引物進行的。
DNA雜交分析植物轉化通過基因組DNA的DNA印跡分析證實(Sambrook和Russell，2001)。通常，從轉化株中提取總DNA，用適當的限制酶消化，在瓊脂糖凝膠中進行分級分離並轉移至硝酸纖維素或尼龍膜上。然後依照標準技術，用例如放射性標記了32P的靶DNA片段探測所述的膜或「印跡」，以證實被導入基因在植物基因組中的整合(Sambrook和Russell，2001.分子克隆實驗室手冊MolecularCloningA Laboratory Manual.冷泉港實驗室出版社，冷泉港，紐約)。
RNA雜交分析依照本領域中常規使用的標準方法，從轉化株的特異組織中分離RNA，在甲醛瓊脂糖凝膠中分級分離，印跡至尼龍濾膜上(Sambrook，J.和Russell，D.W.2001.分子克隆實驗室手冊Molecular CloningA Laboratory Manual.冷泉港實驗室出版社，冷泉港，紐約)。然後用本領域已知方法(Sambrook和Russell，2001)，通過將所述濾膜與來自δ-內毒素或δ-內毒素相關蛋白質的放射性探針雜交而測試δ-內毒素或與δ-內毒素相關基因編碼的RNA的表達。
蛋白質印跡和生化檢測可通過標準步驟(Sambrook，J.和Russell，D.W.2001.分子克隆實驗室手冊Molecular CloningA LaboratoryManual.冷泉港實驗室出版社，冷泉港，紐約)，利用結合δ-內毒素或δ-內毒素相關蛋白質上存在的一個或多個表位的抗體對轉基因植物進行蛋白質印跡和生化檢測及類似測定，以證實由δ-內毒素或δ-內毒素相關基因編碼的蛋白質的存在。
植物中的殺蟲活性在本發明的另一方面，可產生表達具有殺蟲活性的δ-內毒素或δ-內毒素相關蛋白質的轉基因植物。可利用上述舉例說明的方法產生轉基因植物，但產生轉基因植物細胞的方式對本發明而言不是關鍵的。可利用諸如農桿菌介導的轉化、氣溶膠束(aerosol beam)、biolistic轉化及非粒子介導方法等本領域中已知或描述的方法來進行，具體所用方法依實驗者而定。可用本領域中描述的通用方法分離表達δ-內毒素或δ-內毒素相關蛋白質的植物，例如，通過轉化愈傷組織、篩選被轉化的愈傷組織以及從所述的轉基因愈傷組織再生出可育植物。在所述過程中，可利用任何基因作為可選擇標記，只要通過它在植物細胞中的表達能識別或選擇已被轉化的細胞即可。
已開發了許多與植物細胞一起使用的標記物，諸如對氯黴素、氨基糖苷類G418、潮黴素等等的抗性。編碼葉綠體代謝中所涉及的產物的其它基因也可用作選擇標記。例如，能提供對諸如草甘膦、溴苯腈(bromoxynil)或咪唑啉酮(imidazolinone)等植物除草劑的抗性的基因可能有具體用途。這樣的基因已有報導(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)。
可對表達δ-內毒素或δ-內毒素相關蛋白質的可育植物測試殺蟲活性，並選擇顯示出最佳活性的植物進行進一步的培育。本領域中有可供利用的方法能夠用於檢測對害蟲的活性。通常，所述蛋白質被混合併用於餵服試驗中。參閱，例如，Marrone等(1985)J.ofEconomic Entomology 78290-293。
殺蟲控制中的應用將本發明的株系用於殺蟲控制或將其它生物體改造成殺蟲劑的一般方法是本領域所已知的。參閱，例如，美國專利5,039,523和EP0480762A2。
本發明的芽孢桿菌株系或經遺傳改造而含有殺蟲基因和蛋白質的微生物可用於保護農作品和產品免受害蟲的侵襲。在本發明的一個方面，當將產生毒素的細胞施用於有目標害蟲的環境中時，用延長所述細胞中產生的毒素的活性的試劑處理生產毒素(殺蟲劑)的生物體的完整(即未裂解)細胞。
或者，通過將異源基因引入細胞宿主而產生殺蟲劑。異源基因的表達直接或間接的導致了殺蟲劑在細胞內的產生和保持。在本發明的一個方面，當將所述細胞施用於有目標害蟲的環境中時，在延長該細胞所產生的毒素的活性的條件下處理這些細胞。所產生的產物保持了毒素的毒性。然後可依照常規技術配製這些自然封裝的殺蟲劑，以便施用於含有目標害蟲的環境中，如，土壤、水和植物的葉子。參閱，例如EPA 0192319以及其中引用的文獻。或者，可對表達本發明基因的細胞進行配製，以便可將所產生的物質作為殺蟲劑施用。
本發明的活性成分通常以組合物的形式施用，並可與其它化合物同時或接連地施用於待處理的農作物地區或植物。這些化合物可以是肥料、除草劑、防凍劑、表面活性劑、去垢劑、殺蟲皂、休眠油、聚合體和/或在單次施用製劑後可長期對目標區域進行給藥的隨時間釋放型或可生物降解型載體製劑。它們也可以是選擇性除草劑、化學殺蟲劑、殺病毒劑、殺微生物劑、殺變形蟲劑、殺蟲劑、殺真菌劑、殺菌劑、殺線蟲劑、殺軟體動物劑或數種這些製劑的混合物，如果需要，可進一步與農業可接受載體、表面活性劑或製備領域中常規採用的促進應用的佐劑一起施用。合適的載體和佐劑可以是固體或液體，並與製備工藝中通常應用的物質相對應，如，天然或再生的礦物、溶劑、分散劑、增溼劑、粘附劑、粘合劑或肥料。同樣的，所述製劑可製備成可食用的「餌」或作成害蟲「陷阱」，使得可被殺蟲製劑的目標害蟲所飼用或攝取。
應用本發明活性成分或含本發明細菌菌株所產生的殺蟲性蛋白質中至少一種的本發明農用化學品組合物的優選方法是葉片施用、種子包裹和土壤施用。施用次數和施用頻率取決於相應害蟲的侵襲強度。
所述組合物可製備成粉劑、塵劑、丸劑、顆粒劑、噴霧劑、乳劑、膠體、溶液等等，且可用諸如含所述多肽的細胞培養物的乾燥、凍幹、勻漿、提取、過濾、離心、沉降和濃縮等常規方式製備。在所有含至少一種所述殺蟲多肽的所述組合物中，所述多肽可以重量約1％-約99％的濃度存在。
利用本發明方法可殺死鱗翅類或鞘翅類害蟲，或者減少其在特定區域內的數目，或者可預防性的施用於周圍區域以防止易感害蟲的蔓延。優選的，所述害蟲攝取或接觸殺蟲有效量的所述多肽。「殺蟲有效量」指能導致至少一種害蟲死亡或能顯著降低害蟲的生長、飼用或正常生理髮育的殺蟲劑用量。這個量將取決於各種因素而變化，諸如待控制的具體目標害蟲；待處理的特異環境、位置、植物、農作物或農業場所、環境條件，以及該殺蟲有效性多肽組合物施用的方法、速率、濃度、穩定性和用量。製劑還可隨氣候條件、環境因素和/或施用頻率和/或害蟲侵襲的嚴重程度等而發生變化。
所述的殺蟲劑組合物可通過製備細菌細胞、晶體和/或孢子懸浮液或者與所需的農業可接受載體有組合的分離蛋白質組分製成。所述組合物可在施用前配製成適當的形式，諸如凍幹、冷凍乾燥、脫水或在水性載體、培養基或諸如鹽水或其它緩衝液等適當的溶劑中。製備好的組合物可以是塵土或顆粒物形式，或者油(植物油或礦物油)中的懸浮液，或者水或油/水乳劑，或作為可溼粉末，或者與適於農業應用的任何其它載體物質相組合。適當的農業載體可以是固體或液體，在本領域中是眾所周知的。術語「農業可接受載體」涵蓋了通常應用於殺蟲劑製備工藝中的所有佐劑、惰性成分、分散劑、表面活性劑、粘著劑、粘合劑等，這些是殺蟲劑製備業中的技術人員眾所周知的。所述製劑可與一種或多種固體或液體佐劑混合，並通過各種方式製備，例如，用常規的製備工藝將殺蟲組合物與適當的佐劑均勻混合、混和和/或碾磨。合適的製劑和應用方法參閱美國專利6,468,523，該文件在此收編作為參考。
「害蟲」包括但不局限於，昆蟲、真菌、細菌、線蟲、蟎、蜱等等。昆蟲類的害蟲包括選自以下目的昆蟲鞘翅目、雙翅目、膜翅目、鱗翅目、食毛目、同翅目、半翅目、直翅目、纓翅目、革翅目、等翅目、蝨目、蚤目、毛翅目等，尤其是鞘翅目、鱗翅目和雙翅目。
昆蟲類害蟲包括選自以下目的昆蟲鞘翅目、雙翅目、膜翅目、鱗翅目、食毛目、同翅目、半翅目、直翅目、纓翅目、革翅目、等翅目、蝨目、蚤目、毛翅目等，尤其是鞘翅目和鱗翅目。針對主要農作物的本發明昆蟲類害蟲包括玉米Ostrinia nubilalis，歐洲玉米螟；Agrotis ipsilon，小地虎；Helicoverpa zea，谷實夜蛾；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)根葉甲；Diabrotica longicornis barberi，長角葉甲；Diabrotica undecimpunctata howardi，黃瓜十一星葉甲；Melanotus spp.，梳爪叩頭蟲；Cyclocephala borealis，北方圓頭犀金龜(圓頭獨角仙)；Cyclocephala immaculata，南方圓頭犀金龜(圓頭無斑獨角仙)；Popillia japonica，弧麗蛂(日本麗金龜)；Chaetocnema pulicaria，玉米(銅色)跳甲；Sphenophorus maidis，玉米隱喙象(玉米谷象)；Rhopalosiphum maidis，玉米蚜(玉米縊管蚜)；Anuraphis maidiradicis，玉米根蚜；Blissus leucopterusleucopterus，麥長蝽(美洲谷長蝽)；Melanoplus femurrubrum，赤脛黑蝗(赤腿蚱蜢)；Melanoplus sanguinipes，血黑蝗(遷徙蚱蜢)；Hylemya platura，(玉米)種蠅；Agromyza parvicornis，玉米斑潛蠅；Anaphothrips obscrurus，玉米黃呆薊馬(美洲錐尾薊馬)；Solenopsis milesta，竊蟻；Tetranychus urticae，二斑葉蟎(普通紅葉蟎)；高粱Chilo partellus，玉米禾螟；Spodopterafrugiperda，草地夜蛾；Helicoverpa zea，谷實夜蛾；Elasmopalpuslignosellus，南美玉米苗斑螟(小玉米螟)；Feltia subterranea，粒膚地虎；Phyllophaga crinita，蠐螬(金龜幼蟲)；Eleodes，寬胸叩甲(Conoderus)和Aeolus spp.，金針蟲(叩甲幼蟲)；Oulemamelanopus，黑角負泥蟲(橙足負泥蟲)；Chaetocnema pulicaria，玉米(銅色)跳甲；Sphenophorus maidis，玉米隱喙象(玉米谷象)；Rhopalosiphum maidis，玉米蚜(玉米縊管蚜)；Sipha flava，美甘蔗偽毛蚜；Blissus leucopterus leucopterus，麥長蝽(美洲谷長蝽)；Contarinia sorghicola，高粱癭蚊；Tetranychuscinnabarinus，紅葉蟎(硃砂葉蟎)；Tetranychus urticae，葉蟎(普通紅葉蟎)；小麥Pseudaletia unipunctata，粘蟲；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，黑角負泥蟲(橙足負泥蟲)；Hypera punctata，三葉草葉象(車軸草葉象)；Diabrotica undecimpunctata howardi，黃瓜十一星葉甲；俄國麥長管蚜；Schizaphis graminum，麥二岔蚜；Macrosiphumavenae，麥長管蚜；Melanoplus femurrubrum，赤脛黑蝗(赤腿蚱蜢)；Melanoplus differentialis，異黑蝗(殊種蚱蜢)；Melanoplussanguinipes，血黑蝗(遷徙蚱蜢)；Mayetiola destructor，小麥癭蚊；Sitodiplosis mosellana，麥紅吸漿蟲；Meromyza americana，美洲杆蠅(美洲麥杆黃潛蠅)；Hylemya coarctata，麥癭種蠅(冬作種蠅)；Frankliniella fusca，煙褐花薊馬(菸草褐薊馬)；Cephuscinctus，麥莖蜂；Aceria tulipae，麥癭蟎(鬱金香瘤蟎)；向日葵Suleima helianthana，向日葵芽蛀蟲；Homoeosoma electellum，向日葵(同)斑螟；zygogramma exclamationis，向日葵葉甲；Bothyrus gibbosus，胡蘿蔔利犀金龜(胡蘿蔔金龜)；Neolasiopteramurtfeldtiana，向日葵籽癭蚊；棉花Heliothis virescens，煙芽夜蛾；Helicoverpa zea，棉鈴蟲；Spodoptera exigua，甜菜夜蛾；Pectinophora gossypiella，紅鈴麥蛾(棉紅鈴蟲)；Anthonomusgrandis，墨西哥棉鈴象(甲)；Aphis gossypii，棉蚜；Pseudatomoscelis seriatus，棉序盲蝽(棉跳盲蝽)；Trialeurodesabutilonea，苘麻粉蝨(結翅粉蝨)；Lygus lineolaris，(美國)牧草盲蝽；Melanoplus femurrubrum，赤脛黑蝗(赤腿蚱蜢)；Melanoplus differentialis，異黑蝗(殊種蚱蜢)；Thrips tabaci，煙薊馬(蔥薊馬)；Franklinkiella fusca，煙褐花薊馬(菸草褐薊馬)；Tetranychus cinnabarinus，紅葉蟎(硃砂葉蟎)；Tetranychusurticae，二斑葉蟎(普通紅葉蟎)；稻Diatraea saccharalis，小蔗杆草螟；Spodoptera frugiperda，草地夜蛾；Helicoverpa zea，棉鈴蟲；Colaspis brunnea，葡萄肖葉甲；Lissorhoptrus oryzophilus，稻根象；Sitophilus oryzae，米象；Nephotettix nigropictus，黑尾葉蟬；Blissus leucopterus leucopterus，麥長蝽(美洲谷長蝽)；Acrosternum hilare，擬緣椿(喜綠蝽)；大豆Pseudoplusiaincludens，大豆(尺)夜蛾；Anticarsia gemmatalis，梨豆夜蛾；Plathypena scabra，苜蓿綠夜蛾；Ostrinia nubilalis，歐洲玉米螟；Agrotis ipsilon，小地虎；Spodoptera exigua，甜菜夜蛾；Heliothis virescens，煙芽夜蛾；Helicoverpa zea，棉鈴蟲；Epilachna varivestis，墨西哥(大)豆瓢蟲；Myzus persicae，桃蚜；Empoasca fabae，蠶豆微葉蟬；Acrosternum hilare，擬緣蝽(喜綠蝽)；Melanoplus femurrubrum，赤脛黑蝗(赤腿蚱蜢)；Melanoplusdifferentialis，異黑蝗(殊種蚱蜢)；Hylemya platura，(玉米)種蠅；Sericothrips variabilis，大豆牧草蟲；Thrips tabaci，煙薊馬(蔥薊馬)；Tetranychus turkestani，土耳其葉蟎(草莓葉蟎)；Tetranychus urticae，二斑葉蟎(普通紅葉蟎)；大麥Ostrinianubilalis，歐洲玉米螟；Agrotis ipsilon，小地虎；Schizaphisgraminum，麥二岔蚜；Blissus leucopterus leucopterus，麥長蝽(美洲谷長蝽)；Acrosternum hilare，擬緣椿(喜綠蝽)；Euschistusservus，菸草蝽(褐臭蝽)；Delia platura，(灰地)種蠅；Mayetioladestructor，小麥癭蚊；Petrobia latens，麥巖蟎；油籽油菜Brevicoryne brassicae，甘藍蚜(菜蚜)；Phyllotreta cruciferae，十字花跳菜甲；Mamestra configurata，蓓帶夜蛾(披肩夜蛾)；Plutella xylostella，菜蛾；Delia ssp.，地種蠅(花蠅)。
線蟲類包括寄生性線蟲，例如根節、胚囊和損害線蟲，包括異皮線蟲(Heterodera spp.)、根結線蟲(Meloidogyne spp.)和球異皮線蟲(Globodera spp.)，具體而言是胚囊線蟲類，包括但不局限於，Heterodera glycines(大豆異皮線蟲)、Heterodera schachtii(甜菜胞囊線蟲)、Heterodera avenae(燕麥胞囊線蟲)和Globoderarostochiensis(馬鈴薯金線蟲)以及蒼白球異皮線蟲(Globoderapailida)。損害線蟲包括短杆線蟲(Pratylenchus spp.)。
以下實施例僅供例證說明而非起限制作用。
實施例實施例1質粒DNA的抽提以如下方式由菌株ATX 13026或ATX 13002製備質粒DNA。在大量的豐富培養基中培養菌株ATX 13026或ATX 13002的純培養物。將培養液離心以收穫細胞沉澱塊。然後以本領域已知方法用SDS處理細胞沉澱塊，導致細胞壁破損且DNA釋放。然後通過高鹽濃度沉澱蛋白質和大基因組DNA。通過標準乙醇沉澱來沉澱質粒DNA。通過在氯化銫梯度中的高速離心將質粒DNA與任何剩餘的染色體DNA分開。使用UV光顯現梯度中的DNA，並使用注射器吸取較低密度條帶(即較低條帶)。此條帶含有來自菌株(或是ATX 13026或是ATX 13002)的質粒DNA。利用本領域已知方法通過在瓊脂糖凝膠上的顯影來檢查DNA的質量。
實施例2基因的克隆正如本領域已知的，將純化的質粒DNA剪切成5-10kb大小的片段，並且在存在所有四種dNTP的條件下使用T4DNA聚合酶和Klenow片段修補5′和3′單鏈突出。然後，如本領域已知的，通過T4多核苷酸激酶處理而在5′末端添加磷酸根。如本領域已知的，將修復的DNA片段過夜連接到適當製備以接受插入片段(例如通過限制酶消化而生成平端)的標準高拷貝載體(即pBluescript SK+)中。
通過用已知在克隆位點側翼具有切割位點的限制酶消化克隆亞群來分析文庫的質量。確定了高百分比的克隆含有插入片段，平均長度5-6kb。
實施例3文庫平板的高通量測序將通過上述方法製備的文庫塗布在含有適當抗生素的豐富培養基上以維持質粒克隆，並將菌落單獨挑到裝有2ml含有適當抗生素的培養基的96孔板中。將這些板於37℃、以350rpm搖速培養過夜。將板離心而將細胞收集到板底。通過標準的鹼裂解法以高通量方式由這些培養物分離質粒DNA。
以如下方式對來自每塊板的大量克隆測定來自該文庫的克隆的末端序列。使用螢光染料終止物測序技術(Applied Biosystems)和位於克隆位點每一側側翼的標準引物測定選擇用於分析的每個克隆的DNA序列。一旦反應在熱循環儀中完成，則使用標準乙醇沉澱法來沉澱DNA。將DNA重懸於水，並加載到毛細管測序儀上。由克隆位點的任一側測定每個文庫平板的DNA，每個平板的每個插入片段產生兩個閱讀結果。
實施例4測序數據的裝配和篩選將得到的DNA序列彙編成裝配方案，並排列在一起以形成毗連序列群。這可以使用電腦程式有效進行，諸如Vector NTi，或者使用DNA比對和分析程序的Pred/Phrap套餐。將這些毗連序列群連同未能添加到毗連序列群中的任何個別閱讀序列與所有類型的已知殺蟲劑基因的彙編資料庫進行比較。進一步分析鑑定為與已知內毒素或殺蟲劑基因具有同一性的毗連序列群或個別閱讀序列。在得到的序列中，克隆pAX004、pAX006、pAX007、pAX008、pAX009和pAX014含有鑑定為與已知內毒素基因具有同源性的DNA。因此，選擇這些克隆用於進一步測序。
實施例5δ-內毒素基因的測序和鑑定設計與內毒素基因具有同源性的序列退火的引物，使得由這些引物產生的DNA序列將與克隆中的現有DNA序列交疊。這一稱為「寡聚物行走」的方法在本領域是眾所周知的。該方法被用於測定顯示與已知內毒素基因有同源性的區域的完整DNA序列。在上文所述克隆的情況中，該方法被用於測定整個克隆的DNA序列，從而得到每個基因的單一核苷酸序列。然後將完整DNA序列放回最初的大裝配結果中以進一步確認。這能夠將更多的DNA序列閱讀結果插入毗連序列群，產生覆蓋整個區域的多個閱讀結果。
對與已知內毒素基因同源的每個區域進行的DNA序列分析鑑定得到了與已知δ-內毒素基因同源的開放讀碼框。將由pAX004鑑定得到的開放讀碼框命名為AXMI-004。將由pAX006鑑定得到的開放讀碼框命名為AXMI-006。將由pAX007鑑定得到的開放讀碼框命名為AXMI-007。將由pAX008鑑定得到的開放讀碼框命名為AXMI-008。將由pAX009鑑定得到的開放讀碼框命名為AXMI-009。將由pAX014鑑定得到的開放讀碼框命名為AXMI-014。SEQ ID NOS1和2提供了AXMI-004的DNA序列，而SEQ ID NO3提供了預測的AMXI-004蛋白質的胺基酸序列。位於SEQ ID NO1第385位核苷酸處的AXMI-004備選起始位點產生了SEQ ID NO5中提供的胺基酸序列。SEQ ID NOS6提供了AXMI-006的DNA序列，而SEQ ID NO7提供了預測的AMXI-006蛋白質的胺基酸序列。SEQ ID NO8提供了AXMI-007的DNA序列，而SEQ ID NO9提供了預測的AMXI-007蛋白質的胺基酸序列。位於SEQ IDNO8第151位核苷酸處的AXMI-007備選起始位點產生了SEQ ID NO11中提供的胺基酸序列。SEQ ID NOS12和13提供了AXMI-008的DNA序列，而SEQ ID NO14提供了預測的AMXI-008蛋白質的胺基酸序列。位於SEQ ID NO12第177位核苷酸處的AXMI-008備選起始位點產生了SEQ ID NO16提供中的胺基酸序列。進一步的分析鑑定得到了緊臨AXMI-008開放讀碼框3′末端的開放讀碼框。SEQ ID NO18中提供了在本文中稱為AXMI-008orf2的該開放讀碼框的這一預測的胺基酸序列。SEQ ID NOS19中提供了AXMI-009的DNA序列，而SEQ ID NO20中提供了預測的AMXI-009蛋白質的胺基酸序列。位於SEQ ID NO19第34位核苷酸處的AXMI-009備選起始位點產生了SEQ ID NO22中提供的胺基酸序列。位於SEQ ID NO19第64位核苷酸處的另一個AXMI-009備選起始位點產生了SEQ ID NO24中提供的胺基酸序列。SEQ ID NOS25和26中提供了AXMI-014的DNA序列，而SEQ ID NO27提供了預測的AMXI-008蛋白質的胺基酸序列。位於SEQ ID NO25第136位核苷酸處的AXMI-014備選起始位點產生了SEQ ID NO29中提供的胺基酸序列。
實施例6分離基因與已知內毒素基因的同源性用本發明的DNA序列和胺基酸序列搜索DNA和蛋白質資料庫揭示出了這些序列與已知內毒素同源。
AXMI-004圖1顯示了AXMI-004與幾種內毒素的序列比對。Blast搜索確定了cry1Ca具有最強的同源區，在毒性結構域的整體序列同一性為43％(見表1)。
表1.AXMI-004與示範性內毒素類型的胺基酸同一性
AXMI-006圖2顯示了AXMI-006與幾種內毒素的序列比對。Blast搜索確定了cry4Aa具有最強的同源區，儘管AMXI-006蛋白質(SEQ ID NO7)與一大組內毒素蛋白質的序列比對顯示最同源的蛋白質是cry10Aa。cry10Aa與AXMI-006的整體胺基酸同一性是25％(見表2)。對AXMI-006胺基酸序列的檢查提示它不含幾種內毒素家族中存在的C-末端非毒性結構域。由序列比對中除去毒素的這個C-末端蛋白質後，序列比對反映了胺基酸同一性只存在於毒素結構域中(見表2，第3列)。圖2顯示了這一「修剪過的」序列比對。
表2.AXMI-006與示範性內毒素類型的胺基酸同一性
AXMI-007
Blast搜索確定了cry4Aa具有與AXMI-007最強的同源區，儘管AMXI-007蛋白質(SEQ ID NO9)與一大組內毒素蛋白質的序列比對顯示最同源的蛋白質是cry10Aa。cry10Aa與AXMI-007的整體胺基酸同一性是25％(見表3)。對AXMI-007胺基酸序列的檢查提示它不含幾種內毒素家族中存在的C-末端非毒性結構域。由序列比對中除去毒素的這個C-末端蛋白質後，序列比對反映了胺基酸同一性只存在於毒素結構域中(見表3，第3列)。圖3顯示了這一「修剪過的」序列比對。
表3.AXMI-007與示範性內毒素類型的胺基酸同一性
AXMI-008Blast搜索確定了cry40Aa具有與AXMI-008最強的同源區，而且AMXI-008蛋白質(SEQ ID NO14)與一大組內毒素蛋白質的序列比對顯示最同源的蛋白質是cry40Aa。cry40Aa與AXMI-008的整體胺基酸同一性是66％(見表4)。對AXMI-008胺基酸序列的檢查提示它不含幾種內毒素家族中存在的C-末端非毒性結構域。由序列比對中除去毒素的這個C-末端蛋白質後，序列比對反映了胺基酸同一性只存在於毒素結構域中(見表4，第3列)。圖4顯示了這一「修剪過的」序列比對。
表4.AXMI-008與示範性內毒素類型的胺基酸同一性
緊臨AXMI-008編碼區下遊(3′)的開放讀碼框與已知內毒素相關蛋白質具有同源性。Blast搜索確定了crybun3orf2(cry40Aa的下遊開放讀碼框)具有最強的同源區。資料庫中存在幾種其它orf-2樣蛋白質，而且圖5顯示了AMXI-008-orf2蛋白質(SEQ ID NO18)與這些蛋白質中的一組的序列比對。這些蛋白質還與cry4Aa和cry4Ba C-末端非毒性結構域享有同源性。AXMI-8-orf2與cry40Aaorf2的整體胺基酸同一性是86％(見表5)。
表5.AXMI-008-orf2與相關蛋白質的胺基酸同一性
AXMI-009Blast搜索確定了cryBAa具有與AXMI-009最強的同源區，而且AMXI-009蛋白質(SEQ ID NO20)與一大組內毒素蛋白質的序列比對顯示最同源的蛋白質是cry3Ba和cry16Aa。cry3Ba和cry16Aa與AXMI-009的整體胺基酸同一性是26％(見表6)。對AXMI-009胺基酸序列的檢查提示它不含幾種內毒素家族中存在的C-末端非毒性結構域。由序列比對中除去毒素的這個C-末端蛋白質後，序列比對反映了胺基酸同一性只存在於毒素結構域中(見表6，第3列)。圖6顯示了這一「修剪過的」序列比對。
表6.AXMI-009與示範性內毒素類型的胺基酸同一性
AXMI-014Blast搜索確定了cry40Aa具有與AXMI-014最強的同源區，而且AMXI-0014蛋白質(SEQ ID NO27)與一大組內毒素蛋白質的序列比對顯示最同源的蛋白質是cry40Aa。cry40Aa與AXMI-014的整體胺基酸同一性是55％(見表7)。對AXMI-014胺基酸序列的檢查提示它不含幾種內毒素家族中存在的C-末端非毒性結構域。由序列比對中除去毒素的這個C-末端蛋白質後，序列比對反映了胺基酸同一性只存在於毒素結構域中(見表7，第3列)。圖7顯示了這一「修剪過的」序列比對。
表7.AXMI-014與示範性內毒素類型的胺基酸同一性
實施例7AXMI-006與AXMI-007之間的同源性對胺基酸序列AXMI-007與AXMI-006的比較顯示了這兩種毒素共享顯著的胺基酸同源性。胺基酸序列AXMI-006(SEQ ID NO7)和AXMI-007(SEQ ID NO9)的比對顯示了這兩種蛋白質在胺基酸水平上85％相同。因此，AXMI-006和AXMI-007組成一類新的相關內毒素。
實施例8殺蟲活性的測定法常常以多種方式評估殺蟲劑蛋白質作為殺蟲劑作用於害蟲的能力。本領域眾所周知的一種方法是進行餵食測定法。在這種餵食測定法中，將害蟲暴露於含有待測化合物的樣品或對照樣品。通常，這是通過將待測物質或其合適稀釋物置於害蟲將要攝食的物質諸如人工飼料上來進行的。待測物質可以由液體、固體或漿液構成。可以將待測物質置於表面之上，然後使其乾燥。或者，可以將待測物質與熔化的人工飼料混和，然後分裝到測定槽中。測定槽可以是例如杯、盤或微量滴定板的孔。
針對吸食害蟲(例如蚜蟲)的測定法可以包括使用隔離物將測試物質與昆蟲分開，理想的是吸食昆蟲的吸食口器能夠刺穿的物體，從而能夠攝食測試物質。通常，將測試物質與餵食刺激物諸如蔗糖混和以促進測試化合物的攝食。
其它類型的測定法可以包括將測試物質顯微注射到害蟲的口或腸中，以及開發轉基因植物，隨後測試害蟲以轉基因植物為食的能力。植物測試可以包括分離通常取食的植物部分，例如將小籠附於葉片上，或者在裝有昆蟲的籠中分離整個植株。
本領域還知道測定害蟲的其它方法和途徑，可以在如下文獻中找到Robertson，J.L.和H.K.Preisler，1992，《使用節肢動物的殺蟲劑生物測定法》(Pesticide bioassays with arthropods)，CRC，伯克萊屯，佛羅裡達州。或者，測定法一般描述於如下期刊中「Arthropod Management Tests」和「Journal of EconomicEntomology」，或者是與美國昆蟲學學會(Entomological Society ofAmerica，ESA)成員的討論。
實施例9克隆AXMI-006用於蛋白質表達如下將AXMI-006克隆到用於大腸桿菌表達的載體中。pAX480含有用於選擇轉化子的卡那黴素抗性基因，以及可通過IPTG誘導來調控蛋白質表達的tac啟動子。通過將編碼6xHis標籤區的DNA片段緊臨著插入插入片段克隆區的上遊來修飾pAX480，使得得到的克隆在所表達的蛋白質的N端含有6xHis標籤。本領域眾所周知表達含有6xHis標籤融合體的蛋白質的方法及其用於純化和分析蛋白表達的用途。
使用PfuUltraTM高保真DNA聚合酶(Stratagene)PCR擴增AXMI-006的編碼序列。設計寡核苷酸引物，使得得到的PCR產物在兩個末端附近含有期望的限制性位點，以便於克隆。用合適的限制酶消化所得到的PCR產物(約2.2kb)，並亞克隆到經過修飾的pAX480中。通過限制性分析鑑定含有插入片段的克隆。得到的克隆pAX906含有與6xHis標籤融合的AXMI-006開放讀碼框，其轉錄和翻譯導致生成具有一段六個組氨酸的「融合蛋白」。通過DNA序列分析確認了pAX906的DNA序列，隨後如製造商(Stratagene，拉霍亞，加利福尼亞州)所述轉化到化學感受態大腸桿菌BL21中。
將pAX906在BL21中的單菌落接種到添加了卡那黴素的LB培養基中，並於37℃以劇烈振搖培養幾個小時。將這些培養物培養至OD600為0.6-0.8，然後通過添加0.1mM IPTG來誘導蛋白質生成。將培養物在誘導條件下培養3小時，然後通過離心沉澱細胞並重懸於PBS。採用10秒超聲間隔，使用Misonix Sonicator 3000超聲處理細胞，總計30秒，並在冰上保溫1分鐘。
實施例10AXMI-006對煙芽夜蛾和草地夜蛾的活性的生物測定法使用人工飼料(多物種飼料，Southland Products，Lake Village，阿肯色州)進行經過超聲處理的pAX906培養物的生物測定法。通過將經過超聲處理的pAX906細胞或作為對照的大腸桿菌菌株的細胞施加到飼料表面並使飼料表面乾燥來進行生物測定法。在24孔組織培養板中進行生物測定法。生物測定法在黑暗中、25℃和65％相對溼度下進行。用Breathe簡易密封帶(Diversified Biotech，波士頓，麻薩諸塞州)密封盤子。第5天記錄結果。
表8.AXMI-006對草地夜蛾的生物測定法
表9.AXMI-006對煙芽夜蛾的生物測定法
阻礙發育被定義為昆蟲體形變小，而且一些幼蟲攝食減少。發育受阻礙的昆蟲還可能表現出相對於未處理飼料而言迴避經過處理的飼料。
實施例11AXMI-008、AXMI-009和AXMI-014對粉紋夜蛾的殺蟲活性將含有pAX008、pAX009或pAX-014的大腸桿菌菌株，以及未轉化的大腸桿菌的培養物在2ml LB培養基(Luria-Bertani肉湯，BectonDickinson Company，斯帕克斯，馬裡蘭州)中於37℃以250rpm振搖培養24小時。將含有質粒的菌株在含有適當抗生素的LB培養基中培養，以選擇質粒在大腸桿菌中的維持。
使用人工飼料(多物種飼料，Southland Products，Lake Village，阿肯色州)在24孔組織培養板中進行生物測定法。通過將含有pAX-014的大腸桿菌培養物施加到飼料表面並使飼料表面乾燥來進行生物測定法。將菌株的整個培養物以每孔40μl培養物的濃度施加於飼料。生物測定法在黑暗中、25℃和65％相對溼度下進行。用Breathe簡易密封帶(Diversified Biotech，波士頓，麻薩諸塞州)密封盤子。第5天記錄結果。
表10.對粉紋夜蛾的殺蟲活性
實施例12δ-內毒素基因在芽孢桿菌中的表達通過PCR由來自實施例4的克隆擴增AXMI-004、AXMI-006、AXMI-007、AXMI-008和AXMI-009，並通過本領域眾所周知的方法克隆到芽孢桿菌表達載體pAX916中。對於AXMI-004，將得到的克隆命名為pAX920。對於AXMI-006，將得到的克隆命名為pAX921。對於AXMI-007，將得到的克隆命名為pAX919。對於AXMI-008，將得到的克隆命名為pAX922。對於AXMI-009，將得到的克隆命名為pAX917。所得到的克隆在轉化到cry(-)蘇雲金芽孢桿菌菌株的細胞中後表達相應蛋白質。可以在多種常規培養基中培養含有δ-內毒素基因且表達δ-內毒素蛋白質的芽孢桿菌菌株。在CYS培養基(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)中培養含有期望基因的芽孢桿菌菌株，直至通過顯微鏡檢查觀察到孢子形成。製備樣品，並在針對重要昆蟲害蟲的生物測定法中對δ-內毒素蛋白質測試殺蟲活性。
方法配製CYS培養基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混合物應當是pH7，如果需要就進行調節。優選用NaOH或HCl。然後將培養基高壓滅菌，並在高壓滅菌後添加100ml 10X經過濾的葡萄糖。如果得到的溶液渾濁，可以在室溫攪動至清澈。
實施例13在芽孢桿菌中表達的AXMI-004的N端胺基酸序列對於在芽孢桿菌中表達的AXMI-004的分析說明在這些培養物中檢測到的蛋白質產物在長度上相對於全長AXMI-004蛋白質而言可能是縮短的。因為許多內毒素蛋白質在芽孢桿菌中表達後在N端受到切割，我們測定了由芽孢桿菌表達產生的AXMI-004蛋白質的N末端。在PAGE凝膠上分離來自AXMI-004的蛋白質樣品，並且通過本領域眾所周知的方法將蛋白質轉移到PVDF膜上。切下與AXMI-004對應的蛋白質條帶。通過本領域已知的連續Edman降解法測定該蛋白質的N端胺基酸序列。得到的序列如下ERFDKNDALE該胺基酸序列與全長AXMI-004序列(SEQ ID NO3)的比較證明該胺基酸序列來自AXMI-004在芽孢桿菌中表達後的內在切割，導致N末端與SEQ ID NO3第28位胺基酸相對應的蛋白質(公開於SEQ IDNO5)。
實施例14AXMI-004對昆蟲害蟲的生物測定法利用公認的生物測定法流程，使用形成孢子的表達AXMI-004的芽孢桿菌細胞培養物裂解物確定AXMI-004的殺蟲活性。在50ml CYS培養基中培養芽孢桿菌培養物，以用於標準生物測定法和LC50生物測定法。然後將培養物於30℃、以250rpm培養2-3天，直至細胞形成孢子。通過顯微鏡檢查孢子的存在來確定孢子形成。通過將形成孢子的培養物以12,000xg離心10分鐘來製備AXMI-004蛋白質樣品。收集沉澱並重懸於4ml 20mM Tris-HCl，pH8.0。將管置於冰上的同時將懸浮液超聲處理20秒(大功率，使用微型探針)。通過在SDS 4-20％梯度丙烯醯胺凝膠上與已知量的牛血清清蛋白(BSA)一起電泳來測定樣品的蛋白質濃度(圖8)。AXMI-004的濃度經測定是0.4μg/μl。
使用按照本領域所知製備的人工飼料(多物種飼料，SouthlandProducts，Lake Village，阿肯色州)的表面處理生物測定法來測試AXMI-004的殺蟲活性。通過將表達AXMI-004的芽孢桿菌培養物施加到飼料表面並使表面通風乾燥來進行生物測定法。在標準生物測定法中，每個孔使用5個蟲卵，而LC50生物測定法每個孔使用10條新生昆蟲幼蟲。使用細尖毛刷施加蟲卵或幼蟲。標準表面生物測定法在24孔組織培養板中進行。將40ul每種樣品施加到每個孔中。因為每個孔的表面積是2cm2(板的來源)，40μl細胞裂解物樣品含有大約0.4ug/ul AXMI-004。測定LC50的生物測定法在48孔組織培養板中進行，每個孔展示的表面積是1cm2(來源)，每個孔使用大約20ul 0.4μg/ulAXMI-004。每個生物測定法中AXMI-004蛋白質的最終量是大約8μg/cm2。用Breathe簡易密封帶(Diversified Biotech，波士頓，麻薩諸塞州)密封生物測定法的板。對照樣品包括只含培養基的樣品，以及未經樣品處理的孔。然後將生物測定法在黑暗中於25℃和65％相對溼度下放置5天，並記錄結果。
表11.AXMI-004的殺蟲活性
AXMI-004顯示針對歐洲玉米螟和煙芽夜蛾有強殺蟲活性(100％致死)。AXMI-004還顯示針對紅鈴麥蛾有50-75％致死的殺蟲活性。濃度為43μg/cm2的AXMI-004對紅鈴麥蛾產生70％死亡率。AXMI-004嚴重阻礙小地虎、棉鈴蟲和草地夜蛾的發育。
實施例15AXMI-004針對煙芽夜蛾和歐洲玉米螟的殺蟲活性的量化通過在昆蟲生物測定法中測試一定範圍的AXMI-004蛋白質濃度並將這些蛋白質樣品施加到昆蟲飼料表面來測定AXMI-004蛋白質對歐洲玉米螟和煙芽夜蛾幼蟲的LC50。記錄每個蛋白質濃度的死亡率，並使用Probit分析軟體進行分析。結果在95％置信空間是顯著的。因為測定是通過表面汙染來進行的，在假設全部蛋白質樣品在測定過程中都保留在表面上而在昆蟲的攝食水平下很少有擴散的前提下測定LC50。因此，測定的數值可能有些低估AXMI-004蛋白質對測試昆蟲的毒性。
表12.AXMI-004對歐洲玉米螟的LC50
LC50＝297ng/cm2；95％CI＝218-384.
表13.AXMI-004對煙芽夜蛾的LC50
LC50＝2874ng/cm2；95％CI＝2189-3933實施例16AXMI-004、AXMI-006、AXMI-007和AXMI-009針對牧草盲蝽(Lygus lineolaris)的殺蟲活性的量化如下製備細菌裂解物切下芽孢桿菌在50ml CYS培養基中培養60小時。然後將芽孢桿菌培養物以12,000rpm離心10分鐘，並丟棄上清液。將沉澱重懸於5ml 20mM Tris HCl pH8。
如下進行生物測定法切下Eppendorf管的底尖和蓋子，形成餵食槽。將殺蟲劑蛋白質或對照以溶液倒入管帽中並覆蓋昆蟲能夠刺穿從而攝食的石蠟膜(Pechiney Plastic Packaging，芝加哥，伊利諾斯州)從而呈現給昆蟲。將Eppendorf管倒置管帽朝下置於黑暗中，並用細尖毛刷將一齡或二齡Lygus nymphs置於Eppendorf槽中。用石蠟膜密封切下的Eppendorf管尖製成測定槽。將得到的測定槽帽端朝下於環境溫度保溫。在15％葡萄糖溶液中以濃度6.6ug/ml測試殺蟲劑蛋白質。
表14.對牧草盲蝽的殺蟲活性
實施例17AXMI-008對黃粉蟲(Tenebrio molitor)的生物測定法製備表達AXMI-008的芽孢桿菌培養物的樣品，並測試對黃粉蟲的殺蟲活性。當pAX922製備成pH4.0的不溶組分時，它顯示對通常稱為大黃粉蟲的昆蟲有活性。
由含有pAX922的芽孢桿菌菌株的培養物製備AXMI-008的樣品。如下製備生物測定法樣品將培養物在CYS培養基中培養4天，直至孢子形成。將樣品以10,000rpm離心10分鐘，丟棄上清液。用20mMTris pH8清洗沉澱，並以10,000rpm離心10分鐘。丟棄上清液，並將沉澱重懸於3ml 50mM檸檬酸鈉和25mM氯化鈉及2mM DTT pH4。將樣品於37℃保溫1小時。保溫後，將樣品以13,000rpm離心10分鐘，丟棄上清液。將沉澱重懸於50mM檸檬酸鈉和25mM氯化鈉pH4。
使用人工飼料(Southern Corn Rootworm Diet，Bioserv，Frenchtown，新澤西州，#F9757B)在24孔組織培養板中進行樣品對黃粉蟲的生物測定法。通過表面處理以Axmi008濃度8ug/cm2施加樣品，並使之空氣乾燥。使用細尖毛刷施加昆蟲。用Breathe簡易密封帶(Diversified Biotech，波士頓，麻薩諸塞州)密封生物測定盤，並在無光條件下於65％相對溼度和25℃保溫7天並記錄結果。
表15.AXMI-008對黃粉蟲的殺蟲活性
*其餘黃粉蟲發育受到阻礙。發育受阻即觀察到幼蟲體形較小、生長較慢，而且攝食劇烈減少或僅有最低限度。昆蟲還可能表現出相對於未處理飼料而言迴避經過處理的飼料。
實施例18AXMI-009蛋白質對鞘翅類害蟲的生物測定法通過將40μl不溶組分施加到2cm2飼料表面上、達到最終總蛋白質濃度為8μg/cm2來進行AXMI-009蛋白質製劑的生物測定法。使用南方玉米食蟲飼料(Bioserv，Frenchtown，新澤西州，#F9757B)測試玉米根葉甲(Diabrotica virgifera virgifera)和黃瓜十一星葉甲(Diabrotica undecimpunctata)。通過將表達AXMI-009的芽孢桿菌培養物施加到飼料表面並使飼料表面乾燥來進行生物測定法。生物測定法在24孔組織培養板中進行。標準生物測定法每個孔使用25個蟲卵。將蟲卵施加到含有0.1％瓊脂和30ug/ml制黴菌素的溶液中。用Breathe簡易密封帶(Diversified Biotech，波士頓，麻薩諸塞州)密封盤子並將蓋子蓋回盤上。將生物測定法避光於65％相對溼度(RH)和25℃保溫7天。對於玉米根葉甲和黃瓜十一星葉甲都看到了不溶組分的活性。對照是培養基、緩衝液1mM Tris pH10.5、和芽孢桿菌表達載體pAX916。
表16.玉米根葉甲
表17.黃瓜十一星葉甲
實施例19用於植物表達的載體將δ-內毒素編碼區DNA可操作連接用於在植物中表達的合適啟動子和終止子序列。這些序列在本領域是眾所周知的，而且可以包括用於在單子葉植物中表達的水稻肌動蛋白啟動子或玉米泛素啟動子、用於在雙子葉植物中表達的擬南芥UBQ3啟動子或CaMV 35S啟動子、以及nos或PinII終止子。用於構建和鑑定啟動子-基因-終止子構建體的技術在本領域同樣是眾所周知的。
將上文所述植物表達盒與合適的植物選擇標記結合使用，以便於選擇經轉化的細胞和組織，並連接到植物轉化載體中。這些載體可以包括用於由農桿菌介導的轉化的二元載體或用於氣霧劑或biolistic轉化的單一質粒載體。
實施例20玉米細胞的轉化授粉後8-12天採集玉米穗。由穗分離胚，並將大小為0.8-1.5mm的那些胚用於轉化。將胚盾片朝上置於合適的培養基上，諸如DN62A5S培養基(3.98g/L N6鹽；1mL/L(1000x儲液)N6維生素；800mg/LL-天冬醯胺；100mg/L肌醇；1.4g/L L-脯氨酸；100mg/LCasaminoacids；50g/L蔗糖；1mL/L(1mg/mL儲液)2，4-D)，並於25℃在黑暗中保溫過夜。
將由此產生的外植體轉移到篩網上(每塊板30-40粒)，轉移到滲透培養基上達30-45分鐘，然後轉移到光照板上(見例如PCT公開號WO/0138514和美國專利號5,240,842)。
採用基本上如PCT公開號WO/0138514中描述的條件(aerosolbeam accelerator)，使用氣溶膠氣流加速器，將設計在植物細胞中表達δ-內毒素的DNA構建體加速進入植物組織。光照後，將胚在滲透培養基上保溫30分鐘，然後置於保溫培養基上在黑暗中於25℃保溫過夜。為了避免過度損傷經過光照的外植體，在轉移至恢復培養基前將它們至少保溫24小時。再將胚散布到恢復期培養基上，在黑暗中於25℃保溫5天，再轉移至選擇培養基上。將外植體在選擇培養基中保溫可長達8周，這取決於所用具體選擇方法的本質和特徵。選擇期後，將由此產生的愈傷組織轉移至胚成熟培養基，直至觀察到成熟體細胞胚的形成。然後將由此產生的成熟體細胞胚置於低光照下，並通過本領域已知方法啟動再生程序。使由此產生的芽在生根培養基上生根，並將由此產生的植株轉移至苗圃罐中，繁殖成轉基因植物。
材料DN62A5S培養基
用1N KOH/1N KCl將溶液的pH調至pH5.8，添加Gelrite(Sigma)至3g/L，並高壓滅菌。冷卻至50℃後，添加2ml/L硝酸銀(Phytotechnology Labs)的5mg/ml儲液。該配方製備大約20塊平板。
實施例21通過由農桿菌介導的轉化轉化到植物細胞中授粉後8-12天採集穗。由穗分離胚，並將大小為0.8-1.5mm的那些胚用於轉化。將胚盾片朝上置於合適的培養基上，並在黑暗中於25℃保溫過夜。但是，本質上並不必將胚保溫過夜。使胚接觸含有用於Ti質粒介導的轉移的合適載體的農桿菌菌株達5-10分鐘，然後鋪板在共培養培養基上，在黑暗中於25℃保溫3天。共培養後，將外植體轉移至恢復期培養基，在黑暗中於25℃保溫5天。將外植體在選擇培養基中保溫可長達8周，這取決於所用具體選擇方法的本質和特徵。選擇期後，將由此產生的愈傷組織轉移至胚成熟培養基，直至觀察到成熟體細胞胚的形成。然後將由此產生的成熟體細胞胚置於低光照下，並通過本領域已知方法啟動再生程序。使由此產生的芽在生根培養基上生根，並將由此產生的植株轉移至苗圃罐中，繁殖成轉基因植物。
結論正如上文實施例中所示，本發明的δ-內毒素蛋白質針對多種害蟲具有活性。AXMI-004對包括歐洲玉米螟、小地虎、棉鈴蟲、草地夜蛾、煙芽夜蛾、紅鈴麥蛾、菸草天蛾、粉紋夜蛾、和牧草盲蝽在內的害蟲有殺蟲活性。AXMI-006對包括煙芽夜蛾、草地夜蛾、和牧草盲蝽在內的害蟲有殺蟲活性。AXMI-007對包括牧草盲蝽在內的害蟲有殺蟲活性。AXMI-008對包括黃粉蟲和粉紋夜蛾在內的害蟲有殺蟲活性。AXMI-009對包括牧草盲蝽、粉紋夜蛾、玉米根葉甲、和黃瓜十一星葉甲在內的害蟲有殺蟲活性。AXMI-014對包括粉紋夜蛾在內的害蟲有殺蟲活性。
說明書中提到的所有發表物和專利申請指示了本發明所屬領域熟練技術人員的技術水平。本文收錄所有發表物和專利申請作為參考，其程度與專門和個別指出收錄每一項發表物或專利申請作為參考相同。
雖然出於清楚理解的目的通過例示和實施例已經較為詳細的描述了上述發明，顯然可以在所附權利要求的範圍內實踐某些改變和修飾。
序列表110Carozzi，NadineHargiss，TracyKoziel，Michael G.
Duck，Nicholas B.
Carr，Brian120δ-內毒素基因及其使用方法130045600/27437915060/448,6321512003-02-2015060/448,6331512003-02-2015060/448,7971512003-02-2015060/448,8061512003-02-2015060/448,8101512003-02-2015060/448,8121512003-02-2016052170FastSEQ for Windows Version 4.021012112190212DNA213Bacillus thuringiensis4001tatgatgata 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 219021022111890212DNA213Bacillus thuringiensis220
221CDS222(1)...(1890)4002atg 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 *6252103211629212PRT213Bacillus thuringiensis4003Met 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
21042111806212DNA213Bacillus thuringiensis220
221CDS222(1)...(1806)4004atg 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 6002105211601212PRT213Bacillus thuringiensis4005Met 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 60021062112208212DNA213Bacillus thuringiensis220
221CDS222(1)...(2208)4006atg 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 7352107211735212PRT213Bacillus thuringiensis4007Met 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 7352108
2112235212DNA213Bacillus thuringiensis220
221CDS222(1)...(2235)4008gtg 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 *7402109211744212PRT213Bacillus thuringiensis4009
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 Asp740210102112085212DNA213Bacillus thuringiensis220
221CDS222(1)...(2085)40010atg 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 *69021011211694212PRT213Bacillus thuringiensis40011Met 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 Asp690210122115980212DNA213Bacillus thruingiensis40012tacatgcaat 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 5980210132112082212DNA213Bacillus thuringiensis220
221CDS222(1)...(2082)40013gtg 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 *69021014211693212PRT213Bacillus thuringiensis
40014Met 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 Asn690210152112073212DNA213Bacillus thuringiensis220
221CDS222(1)...(2073)40015atg 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 *69021016211690212PRT213Bacillus thuringiensis40016Met 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 Asn690210172111686212DNA213Bacillus thuringiensis220
221CDS222(1)...(1686)40017gtg 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 *21018211561212PRT213Bacillus thuringiensis40018Met 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 560Arg210192112049212DNA
213Bacillus thuringiensis220
221CDS222(1)...(2049)40019atg 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 68021020211682212PRT213Bacillus thuringiensis40020Met 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
210212112016212DNA213Bacillus thuringiensis220
221CDS222(1)...(2016)A0021atg 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 67021022211671212PRT213Bacillus thuringiensis40022Met 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
210232111986212DNA213Bacillus thuringiensis220
221CDS222(1)...(1986)40023atg 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 *66021024211661212PRT213Bacillus thuringiensis40024Met 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
210252112145212DNA213Bacillus thuringiensis40025tttcaccaaa 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 2145210262112019212DNA213Bacillus thuringiensis220
221CDS222(1)...(2019)40026gtg 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*21027211672212PRT213Bacillus thuringiensis40027Met 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 670210282112010212DNA213Bacillus thuringiensis220
221CDS222(1)...(2010)40028atg 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 66521029211669212PRT213Bacillus thuringiensis40029Met 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 665210302111176212PRT213Bacillus thuringiensi40030Met 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 Glu1170117521031
2111178212PRT213Bacillus thuringiensi40031Met 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 Glu11701175210322111189212PRT213Bacillus thuringiensi40032Met 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 Glu118521033211719212PRT213Bacillus thuringiensi40033Met 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 71521034211633212PRT213Bacillus thuringiensi40034Met 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 63021035211652212PRT213Bacillus thuringiensi40035Met 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 65021036211659212PRT213Bacillus thuringiensi40036Met 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
21037211652212PRT213Bacillus thuringiensi40037Met 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 650210382111180212PRT213Bacillus thuringiensi40038Met 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 Glu117011751180210392111136212PRT213Bacillus thuringiensi40039Met 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 Glu11251130113521040211475212PRT213Bacillus thuringiensi40040Met 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 475210412111138212PRT213Bacillus thuringiensi40041Met 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 Cys210422111157212PRT213Bacillus thuringiensi40042Met 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 Glu115521043211675212PRT213Bacillus thuringiensi40043Met 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 Asn67521044211648212PRT213Bacillus thuringiensi40044Met 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
64521045211682212PRT213Bacillus thuringiensi40045Met 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 68021046211529212PRT213Bacillus thuringiensi40046Val 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 525Glu21047211674212PRT213Bacillus thuringiensi40047Met 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 Asn21048211675212PRT213Bacillus thuringiensi
40048Met 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 Asn67521049211659212PRT213Bacillus thuringiensi40049Asn 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 Asn21050211558212PRT213Bacillus thuringiensi40050Met 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 55521051211666212PRT213Bacillus thuringiensi40051Asn 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 66521052211558212PRT213Bacillus thuringiensi40052Met 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權利要求
1.選自下組的分離核酸分子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)含有與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編碼具有殺蟲活性的多肽；c)編碼含有SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示胺基酸序列的多肽的核酸分子；d)含有編碼具有與SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示胺基酸序列有至少95％胺基酸序列同一性的多肽的核苷酸序列的核酸分子，其中所述多肽具有殺蟲活性；和e)a)-d)中任一項的互補物。
2.權利要求1的分離核酸分子，其中所述核苷酸序列是設計成在植物中表達的合成序列。
3.權利要求2的核酸分子，其中所述合成序列具有升高的GC含量。
4.含有權利要求1的核酸分子的載體。
5.權利要求4的載體，其中還含有編碼異源多肽的核酸分子。
6.含有權利要求4的載體的宿主細胞。
7.權利要求6的宿主細胞，它是細菌宿主細胞。
8.權利要求6的宿主細胞，它是植物細胞。
9.含有權利要求8的宿主細胞的轉基因植物。
10.權利要求9的轉基因植物，其中所述植物選自下組玉米、高粱、小麥、向日葵、番茄、十字花科植物、胡椒屬植物、馬鈴薯、棉花、稻、大豆、甜菜、甘蔗、菸草、大麥、和油籽油菜。
11.權利要求9的植物的轉基因種子。
12.選自下組的分離多肽a)含有SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示胺基酸序列的多肽；b)由SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28所示核苷酸序列編碼的多肽，其中所述多肽具有殺蟲活性；c)含有與SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示胺基酸序列具有至少95％序列同一性的胺基酸序列的多肽，其中所述多肽具有殺蟲活性；和d)由與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％相同的核苷酸序列編碼的多肽。
13.權利要求12的多肽，其中還含有異源胺基酸序列。
14.選擇性結合權利要求12的多肽的抗體。
15.含有權利要求12的多肽的組合物。
16.權利要求15的組合物，其中所述組合物選自下組粉劑、塵劑、丸劑、顆粒劑、噴霧劑、乳液、膠體和溶液。
17.權利要求15的組合物，其中所述組合物是通過對蘇雲金芽孢桿菌細胞進行乾燥、凍幹、勻漿、抽提、過濾、離心、沉降、或濃縮而製備的。
18.權利要求15的組合物，其中含有約1重量％至約99重量％的所述多肽。
19.用於生產具有殺蟲活性的多肽的方法，其中包括在編碼所述多肽的核酸分子得到表達的條件下培養權利要求6的宿主細胞，所述多肽選自下組a)含有SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示胺基酸序列的多肽；b)由SEQ ID NO1、2、4、6、8、10、12、13、15、17、19、21、23、25、26或28所示核苷酸序列編碼的多肽，其中所述多肽具有殺蟲活性；c)含有與SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27或29所示胺基酸序列具有至少95％序列同一性的胺基酸序列的多肽，其中所述多肽具有殺蟲活性；和d)由與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％相同的核苷酸序列編碼的多肽。
20.用於控制鱗翅類或鞘翅類害蟲種群的方法，其中包括使所述種群接觸殺蟲有效量的權利要求12的多肽。
21.用於殺死鱗翅類或鞘翅類害蟲的方法，其中包括使所述害蟲接觸或給其餵食殺蟲有效量的權利要求12的多肽。
22.在其基因組中穩定摻入了DNA構建體的植物，其中所述DNA構建體含有編碼具有殺蟲活性的蛋白質的核苷酸序列，該核苷酸序列選自下組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)與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編碼具有殺蟲活性的多肽；c)編碼含有SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29所示胺基酸序列的多肽的核苷酸序列；和d)編碼與SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29所示胺基酸序列具有至少95％胺基酸序列同一性的多肽的核苷酸序列，其中所述多肽具有殺蟲活性；其中所述核苷酸序列可操作性連接了驅動編碼序列在植物細胞中表達的啟動子。
23.權利要求22的植物，其中所述植物還含有SEQ ID NO17所示核苷酸序列。
24.在其基因組中穩定摻入了DNA構建體的植物細胞，其中所述DNA構建體含有編碼具有殺蟲活性的蛋白質的核苷酸序列，該核苷酸序列選自下組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)與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編碼具有殺蟲活性的多肽；c)編碼含有SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29所示胺基酸序列的多肽的核苷酸序列；和d)編碼與SEQ ID NO3、5、7、9、11、14、16、20、22、24、27或29所示胺基酸序列具有至少95％胺基酸序列同一性的多肽的核苷酸序列，其中所述多肽具有殺蟲活性；其中所述核苷酸序列可操作性地連接了驅動編碼序列在植物細胞中表達的啟動子。
25.權利要求24的植物細胞，其中所述植物細胞還含有SEQ IDNO17所示核苷酸序列。
全文摘要
本發明提供了賦予細菌、植物、植物細胞、組織和種子以殺蟲活性的組合物和方法。本發明提供了含有δ－內毒素和δ－內毒素相關多肽的編碼序列的組合物。所述編碼序列可用於DNA構建體或表達盒中，以便用於植物和細菌中的轉化和表達。組合物還包括被轉化的細菌、植物、植物細胞、組織和種子。具體而言，本發明提供了分離的δ－內毒素和δ－內毒素相關核酸分子。另外，本發明還涵蓋與多核苷酸對應的胺基酸序列。具體而言，本發明提供了含有編碼SEQ ID NO3、5、7、9、11、14、16、18、20、22、24、27和29中所示胺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離的核酸分子及其變體和片段。
文檔編號C07K16/12GK1761753SQ200480007563
公開日2006年4月19日 申請日期2004年2月20日 優先權日2003年2月20日
發明者N·卡羅奇, T·哈吉斯, M·G·科奇埃爾, N·B·達克, B·卡爾 申請人:阿則耐克斯公司