CN106928329B - 一种新的杀虫蛋白及其核苷酸序列 - Google Patents
一种新的杀虫蛋白及其核苷酸序列 Download PDFInfo
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Abstract
本发明涉及一种新的杀虫蛋白及其核苷酸序列。杀虫蛋白具有如(I)和(II)中的至少一种的氨基酸序列：(I)SEQ ID No.2所示的氨基酸序列；(II)与如SEQ ID No.2所示的氨基酸序列的一致性在98％以上，且与如SEQ ID No.2所示的氨基酸序列具有相同功能的氨基酸序列；优选其氨基酸序列为与如SEQ ID No.2所示的氨基酸序列的一致性在99.5％以上，且与如SEQ ID No.2所示的氨基酸序列具有相同功能的氨基酸序列。
Description
技术领域
本发明涉及一种新的杀虫蛋白及其核苷酸序列。
背景技术
虫害是造成农作物减产的重要原因之一，减少虫害的损失是增加粮食与饲料作物产量的重要途径。据统计全球粮食与饲料作物总产量每年因虫害造成的损失达14％，直接给农业生产造成的经济损失高达数千亿美元。我国每年因虫害造成的损失水稻减产10％、小麦减产20％、棉花减产30％以上。
其中，棉铃虫(Helicoverpa armigera)和玉米螟(Ostrinia furnacalis)是两种广泛分布于世界各地重要的农业害虫。
棉铃虫属鳞翅目夜蛾科，寄主很广，已知的寄主便有200多种，在蔬菜上主要危害茄科和豆类，其中以番茄和辣椒受害最重，除此外还危害十字花科蔬菜和瓜类。在作物上是棉花、花生、大豆上的重要害虫，并且还可危害小麦、玉米、高粱、豆类、烟草、芝麻、向日葵、苹果、梨、桃、葡萄等。多食性，区域性灾变是棉铃虫发生的特点，对它的防控是我国农业生产面对的长期问题，上世纪棉铃虫危害棉花造成了巨大的经济损失，当时棉铃虫的防治还主要依赖化学药剂，对环境和人畜造成了严重的危害，以及产生了严重的抗药性问题。
玉米螟(Ostrinia furnacalis)俗名钻心虫，属鳞翅目螟蛾科，是我国玉米生产上最重要的害虫，低龄幼虫危害玉米心叶，大龄幼虫钻蛀危害玉米茎秆、雄穗柄、雌穗柄和雌穗，每年因亚洲玉米螟的危害造成的产量损失为600万到900万吨。
采用喷施化学农药和生物杀虫剂等防治手段固然可以减轻害虫对农作物的为害，但化学农药造成环境污染，生物杀虫剂成本较高。长期以来，大量喷施化学杀虫剂，不仅会增强害虫的抗药性，使益虫及其它生态区系遭受破坏，而且严重污染环境，提高生产成本，破坏生态平衡。因此，减少杀虫剂使用量，发展现代植物保护技术，已成为可持续发展农业中必须正视的课题之一。
苏云金芽胞杆菌(Bacillus thuringiensis，简称Bt)是一种分布广泛的革兰氏阳性细菌，是一种对害虫毒力强且对天敌无毒性的昆虫病原微生物，对高等动物和人无毒性。它是目前研究最为深入、使用最为广泛的微生物杀虫剂，对16个目3000多种害虫有活性。Bt在芽胞形成期可形成杀虫晶体蛋白(Insecticidal CrystalProteins，ICPs)，也称δ-内毒素(delta-endotoxin)[Bravo.A.,GillS.S.,
M.BacillusthuringiensisMechanisms and Use.Comprehensive Molecular InsectScienceElsevier,2005,175～206.]，它的形状、结构和大小均与其毒力有着密切关系。苏云金芽胞杆菌杀虫晶体蛋白因其杀虫效果好、对人畜无害[De Maagd R.A.,Bravo A.,CrickmoreN.How Bacillus thuringiensis has evolved specific toxins to col onize theinsect world.Trends Genet,2001,17:193～199.]，不污染环境，因而Bt在害虫的生物防治中得到了最广泛的应用。
1996年全世界第一例转基因抗虫植物在美国获准应用，它使用的基因来自Btcry1Ac。在接下来的几年里，转cry1Ab基因的抗虫玉米，转cry3Aa基因的抗虫土豆等相距问世。在中国，自1998年开始正式推广含有cry1Ac/cry1A基因的抗虫棉以来，已经被普遍种植，棉铃虫的危害也得到了很好的控制。从1996年到2015年的20年期间全球转基因作物累计种植面积达到空前的20亿公顷，相当于中国大陆总面积(9.56亿公顷)或美国总面积(9.37亿公顷)的2倍。这累计的20亿公顷包括：10亿公顷转基因大豆、6亿公顷转基因玉米、3亿公顷转基因棉花和1亿公顷转基因油菜。20年间，农民获益超过1500亿美元。二十年的商业化证明，转基因作物已经实现了其先前的承诺，为农民乃至为全社会带来了农业、环境、经济、健康和社会效益。转基因作物的快速应用表明，那些进行了转基因作物商业化种植的发达国家的大农场主和发展中国家的小农户都已经认识到这种巨大的多重收益。
然而，由于目前商品化的转基因抗虫作物的抗虫基因种类比较单一，如此大面积推广种植存在害虫避难所减少与害虫抗药性上升的风险。因此需要不断分离更高毒力的基因或者新的基因组合来避免害虫抗药性上升的风险。
发明内容
为此，本申请之一提供了一种杀虫蛋白，其具有如(I)和(II)中的至少一种的氨基酸序列：
(I)SEQ ID No.2所示的氨基酸序列；
(II)与如SEQ ID No.2所示的氨基酸序列的一致性在98％以上，且与如SEQ IDNo.2所示的氨基酸序列具有相同功能的氨基酸序列。
通过(II)限定的杀虫蛋白在与如SEQ ID No.2所示的氨基酸序列具有相同功能的同时，也具有与如SEQ ID No.2所示的氨基酸序列相当或者更优的效果，因此可以明显地除去已经被公开的杀虫蛋白。
其中，全长杀虫晶体蛋白可以经过蛋白酶等作用的切割，切割得到约52-65kDa(例如具有55-60kDa)的蛋白后能够更好地发挥其活性。因此，具有如(II)的氨基酸序列的所述的杀虫蛋白可以是具有如(I)的氨基酸序列经过胰凝乳蛋白酶和/或胰蛋白酶消化后获得的具有52-65kDa的蛋白，且与如SEQ ID No.2所示的氨基酸序列具有相同功能的氨基酸序列；优选具有如(II)的氨基酸序列的所述的杀虫蛋白可以是具有如(I)的氨基酸序列经过胰凝乳蛋白酶和/或胰蛋白酶消化后获得的具有55-60kDa的蛋白，且与如SEQ ID No.2所示的氨基酸序列具有相同功能的氨基酸序列。
所述的切割可以一般发生在有害生物的肠道之内，但也可以人为地有目的地改造为具有相同功能的52-65kDa(例如具有55-60kDa)的蛋白。
在一个具体实施例中，优选杀虫蛋白的氨基酸序列为与如SEQ ID No.2所示的氨基酸序列的一致性在99％以上，且与如SEQ ID No.2所示的氨基酸序列具有相同功能的氨基酸序列。通过这种方式限定的杀虫蛋白在与如SEQ ID No.2所示的氨基酸序列具有相同功能的同时，也具有与如SEQ ID No.2所示的氨基酸序列相当或者更优的效果，因此可以明显地除去已经被公开的杀虫蛋白。
在一个具体实施例中，优选杀虫蛋白的氨基酸序列为与如SEQ ID No.2所示的氨基酸序列的一致性在99.5％以上，且与如SEQ ID No.2所示的氨基酸序列具有相同功能的氨基酸序列。通过这种方式限定的杀虫蛋白在与如SEQ ID No.2所示的氨基酸序列具有相同功能的同时，也具有与如SEQ ID No.2所示的氨基酸序列相当或者更优的效果，因此可以明显地除去已经被公开的杀虫蛋白。
在一个具体实施例中，所述杀虫蛋白还可以为如(I)的氨基酸序列经过取代和/或缺失和/或添加一个或多个氨基酸后，与具有如(I)的氨基酸序列的杀虫蛋白具有相同功能的蛋白。通过这种方式限定的杀虫蛋白在与如SEQ ID No.2所示的氨基酸序列具有相同功能的同时，也具有与如SEQ ID No.2所示的氨基酸序列相当或者更优的效果，因此可以明显地除去已经被公开的杀虫蛋白。
本申请的杀虫蛋白对害虫体重的抑制活性，特别是对棉铃虫和玉米螟的体重抑制活性显著优于现有技术中的杀虫蛋白的抑制活性，因此，其不但丰富杀虫基因资源，为转基因作物与工程菌株提供新的基因来源，提高Bt转基因产品的抗虫效果，并且可以降低害虫对Bt毒蛋白的抗性风险，避免新的生态灾难降临，具有重要的经济、社会和生态效益。杀虫蛋白对棉铃虫和玉米螟等害虫具有的优良的体重抑制作用使得大多数害虫不能完成其生长发育，不但会大大减少害虫的取食量，达到了保护植物的目的；而且即使有小部分可以发育为成虫，其产生的后代的质量和数量都会受到影响，比如其后代容易产生某些缺陷。从某种程度上来讲，这种对害虫的体重抑制作用一方面有利于延缓害虫的抗性，另一方面对害虫产生的后代不利，从而减弱其整个种群在环境中的优势地位，属于不可多得的防治害虫的植保材料。
本申请之二提供了一种核苷酸序列，所述核苷酸序列为编码如本申请之一所述的杀虫蛋白中任意一种的核苷酸序列。该核苷酸序列可以以其来源菌株为模板，或以克隆的全长DNA片段为模板，通过PCR扩增得到；也可以根据给定的序列人工合成。
在一个具体的实施例中，当所述杀虫蛋白的氨基酸序列如SEQ ID No.2所示时，编码所述杀虫蛋白的所述核苷酸序列如SEQ ID No.1所示。
本申请之三提供了一种组合物，其包括如本申请之一所述杀虫蛋白的至少一种。
在一个具体的实施例中，当所述杀虫蛋白的氨基酸序列如SEQ ID No.2所示时，编码所述杀虫蛋白的核苷酸序列如SEQ ID No.1所示。
本申请之四提供了一种含有本申请之二所述的核苷酸序列，且能产生本申请之一相应的所述的杀虫蛋白的转基因微生物。
在一个具体的实施例中，优选所述转基因微生物包括芽胞杆菌(Bacillus)、假单胞菌(Pseudomonas)、肠杆菌(Escherichia)和酵母(Saccharomyces)中的至少一种。
在一个具体的实施例中，优选所述芽胞杆菌包括苏云金芽胞杆菌(Bacillusthuringiensis)、枯草芽胞杆菌(Bacillus subtilis)、萎缩芽胞杆菌(Bacillusatrophaeus)和蜡样芽胞杆菌(Bacillus cereus)中的至少一种；所述假单胞菌包括荧光假单胞杆菌(Pseudomonas fluorescens)；所述肠杆菌包括大肠杆菌(Escherichia coli)。
在一个具体实施例中，本申请的转基因微生物的出发株(即在转入本申请之二所述的核苷酸序列之前的微生物)为野生微生物和/或遗传工程微生物。
其中，野生微生物是指从自然界中分离出的未经过人工改造的微生物。
遗传工程微生物是指将野生微生物人工改造的微生物。
本申请的转基因微生物和/或遗传工程微生物的物种并不因进行了转基因修饰而改变，因此，转基因微生物与发生转基因之前的微生物(即作为受体微生物)为相同的物种。
在一个具体实施例中，如上的本申请二的所述的核酸序列能够被连接在表达载体上。所述表达载体例如可以是能够在大肠杆菌和苏云金芽胞杆菌中穿梭的pSTK表达载体。
本申请之五提供了根据本申请之二所述的核苷酸序列在制备转基因植物中的应用，其中，所述转基因植物能够产生如本申请之一所述的杀虫蛋白；所述转基因植物包括禾本科(Liliaceae)、豆科(Leguminosae)、藜科(Chenopodiaceae)、菊科(Compositae)、和锦葵科(Malvaceae)中的至少一种。
在一个具体的实施例中，优选所述转基因植物包括蜀黍属(Zea)、高梁属(Sorghum)、狗尾巴草属(Setaria)、稻属(Oryza)、甘蔗属(Saccharum)、小麦属(Triticum)、大豆属(Glycine)、苜蓿属(Medicago)、甜菜属(Beta)、向日葵属(Helianthus)和棉属(Gossypium)中的至少一种。
在一个具体的实施例中，优选所述转基因植物包括玉米(Zea mays L.)、高粱(Sorghum bicolor(L.)Moench)、谷子(Setaria italica)、水稻(Oryza sativa L.)、甘蔗(Saccharum officinarum)、小麦(Triticum aestivum)、大豆(Glycine max(L.)Merr)、苜蓿(Medicago sativa Linn)、甜菜(Beta vulgaris)、向日葵(Helianthus)、亚洲棉(Gossypium arboreum L.)、非洲棉(Gossypium herbaceum L.)、陆地棉(Gossypiumhirsutum L.)和海岛棉(Gossypium barbadense L.)中的至少一种。
转基因植物的物种并不因进行了转基因修饰而改变，因此，转基因植物与发生转基因之前的植物(即作为受体植物)为相同的物种。
本申请之六提供了一种制备如本申请之一所述的杀虫蛋白的方法，其包括利用本申请之四所述的转基因微生物，和/或利用本申请之五所述的应用中的转基因植物来生产所述杀虫蛋白。
在一个具体的实施例中，优选还包括在利用所述转基因微生物和/或转基因植物生产所述杀虫蛋白后，纯化所述杀虫蛋白使所述杀虫蛋白的纯度达到80％以上。
在一个具体的实施例中，更优选在利用所述转基因微生物和/或转基因植物生产所述杀虫蛋白后，纯化所述杀虫蛋白使所述杀虫蛋白的纯度达到90％以上。
在一个具体的实施例中，甚至是在利用所述转基因微生物和/或转基因植物生产所述杀虫蛋白后，纯化所述杀虫蛋白使所述杀虫蛋白的纯度达到99％以上。
因此，本申请之七提供了根据本申请之一所述的杀虫蛋白，本申请之三所述的组合物，本申请之四所述的转基因微生物，以及本申请之五所述的应用中的转基因植物中的至少一种在防治鳞翅目(Lepidoptera)中的至少一种害虫中的应用。
在一个具体的实施例中，优选根据本申请之一所述的杀虫蛋白，本申请之三所述的组合物，本申请之四所述的转基因微生物，以及本申请之五所述的应用中的转基因植物中的至少一种在防治夜蛾科(Noctuidae)和螟蛾科(Pyralidae)中的至少一种害虫中的应用。
在一个具体的实施例中，特别优选根据本申请之一所述的杀虫蛋白，本申请之三所述的组合物，本申请之四所述的转基因微生物，以及本申请之五所述的应用中的转基因植物中的至少一种在防治铃夜蛾属(Helicoverpa)和秆野螟属(Ostrinia)中的至少一种害虫中的应用。
在一个具体的实施例中，最优选根据本申请之一所述的杀虫蛋白，本申请之三所述的组合物，本申请之四所述的转基因微生物，以及本申请之五所述的应用中的转基因植物中的至少一种在防治棉铃虫(Helicoverpa armigera Hubner)和/或玉米螟(Ostrinianubilalis)中的应用。
单克隆抗体的制备技术已经较为成熟，因此，根据现有技术的常规技术手段，可以制得本申请之一所述的杀虫蛋白的单克隆抗体。因此，本申请之八提供了根据本申请之一所述的杀虫蛋白的单克隆抗体。
在本申请中没有特殊说明的情况下，本申请中的术语均属于现有技术中所指的通用术语。
具体实施方式
以下通过优选的实施例的形式对本发明的上述内容再作进一步的详细说明，但不构成对本发明的限制。
1)液体LB：胰蛋白胨1％，酵母粉0.5％，NaCl 1％，pH 7.0，15磅灭菌15min。用于培养大肠杆菌。
2)固体LB：在液体LB培养基中加1.3％琼脂，15磅灭菌15min。用于培养大肠杆菌。
3)抗生素：氨苄青霉素水溶液100mg/ml，用时稀释500倍-20℃保存。其中pET21b质粒载体具有氨苄青霉素抗性。
蛋白电泳检测
120V预电泳约10-20min，取蛋白样品40μL，加入10μL 5×加样缓冲液，混匀，再将上清和沉淀分别梯度稀释，煮沸5-10min，12000rpm离心5min，取10L上清点样。80V恒压电泳至样品浓缩呈直线，150V恒压电泳至指示的溴酚蓝到达凝胶底部。
脱色：电泳后取出凝胶，电泳后取出凝胶并用蒸馏水冲洗，加入50mL溶液Ⅰ，微波炉中加热30s，60rpm振荡10min。
染色：倒掉溶液I后加入溶液II(每50mL溶液II加200L溶液III)微波炉加热30s，60rpm振荡15min以上。
倒掉溶液II，加入无菌水，凝胶成像系统照相保存。
观察结果，根据蛋白条带着色程度比较目的蛋白表达情况，用凝胶成像系统照相保存。
表1为SDS聚丙烯酰氨凝胶的制备表。
表1SDS聚丙烯酰氨凝胶的制备表
实施例1
新基因的筛选及克隆
PCR-RFLP鉴定：对实验室保存的2000株苏云金芽胞杆菌(Bacillusthuringiensis，Bt)菌株进行基因组提取、定量、均一化后，等量混合，建立Bt基因组池，并以其为PCR模板，利用引物S5un2(SEQ ID No.3)：GGAAGAACTACTATTTGTGATGC；S3un2(SEQ IDNo.4)：AATAGTTTGAATTACCGCGAGC，进行PCR扩增。扩增循环：94℃变性1min；54℃退火1min；72℃延伸2min20s；30个循环，最后72℃延伸10min，PCR产物用0.7％琼脂糖凝胶电泳检测。然后对PCR产物进行RFLP酶切，结合对PCR产物的测序分析，发现了一种新的基因，暂将其定名为cry2A-like。
新基因的克隆与表达：以建立Bt基因组池为模板，利用引物cry2F：5‘-CCGGAATTCgATGAATAATGTATTG-3’；cry2R：5‘-CCCAAGCTTATAAAGTGGTGGAAG-3’，采用pfu高保真聚合酶进行PCR扩增。回收PCR产物，用限制酶EcoR I和Hind III处理pET21b载体和回收的PCR产物，然后分别回收酶切产物，将目的片段连接到载体上，命名为pETcry2A-like，并转化大肠杆菌JM110菌株，筛选出阳性克隆。最后将pETcry2A-like重组质粒转化之大肠杆菌表达菌株Rosetta(DE3)中。
经测序分析，cry2A-like(SEQ ID No.1)基因长度1899bp，其编码蛋白为Cry2A-like，共633个氨基酸(SEQ ID No.2)，表达71.1kDa蛋白。
将上述基因在大肠杆菌Rosetta(DE3)中进行IPTG诱导表达，SDS-PAGE电泳结果发现，cry2A-like基因在可溶组分和不可溶组分中均表达；其中表达约70kDa左右蛋白，与预期结果相符。阴性对照(即不含有cry2A-like基因的大肠杆菌Rosetta(DE3))在可溶和不可溶性组份中都没有相应蛋白的表达条带。
实施例2
蛋白的表达与定量分析
将携带cry2A-like基因的大肠杆菌Rosetta(DE3)菌株以1％的接种量接种于LB液体培养基中，37℃培养至OD600值达到0.5-1.0之间时，加入诱导物50mM IPTG，在150rpm，20℃低温下诱导12h。然后在4℃下，8000rpm离心3min。离心收集菌体，加入50mM Tris·Cl(pH8.0)悬浮；破碎菌体(利用超声波常规完全破碎)，将超声破碎后的菌液在4℃下12,000rpm离心15min；然后收集上清液。按照如下方法对所述上清液进行SDS-PAGE定量分析：制备以下5种不同浓度梯度的BSA：0.8μg/μL、0.4μg/μL、0.2μg/μL、0.1μg/μL、0.05μg/μL，并将目标蛋白进行梯度稀释，使其终浓度介于0.8-0.05μg/μL，目标蛋白和5种不同浓度的BSA(蛋白定量试剂盒：DC Protein Assay)上样量均为10μL，进行蛋白电泳检测。以牛血清蛋白(BSA)为标准蛋白制作蛋白浓度标准曲线，测定大肠杆菌Rosetta(DE3)重组菌株中总蛋白的浓度。结合ImageMaster VDS软件分析SDS-PAGE中目的蛋白所占比例，计算目的蛋白，即Cry2A-like的浓度。
对比例1
以CN201310150965.4中克隆到pET21b上的cry2Ah-likesp(SEQ ID No.7)基因在大肠杆菌Rosetta(DE3))中相同条件下的表达产物为生物活性测定分析的阳性对照。Cry2Ah-likesp(SEQ ID No.8)蛋白的表达与定量分析同实施例2。
实施例3
活性分析
敏感品系棉铃虫(Helicoverpa armigera)、抗Cry1Ac 1000倍的抗性品系棉铃虫(Helicoverpa armigera)和敏感品系亚洲玉米螟(Ostrinia furnacalis)分别由中国农科院植物保护研究所棉花害虫组和玉米害虫组提供的标准化试虫。
棉铃虫饲料由中国农科院植物保护研究所棉花害虫组提供；亚洲玉米螟饲料由中国农科院植物保护研究所玉米害虫组提供。
校正抑制率(％)＝1-处理组平均体重/阴性对照组平均体重
利用POLO Plus软件算出蛋白对试虫的抑制中浓度(IC50)。
1.敏感品系棉铃虫的室内杀虫活性测定
根据实施例2中的杀虫蛋白的定量结果，首先将杀虫蛋白进行系列稀释；然后称取8份10g人工饲料中分别置于灭菌培养皿中，并向每份的人工饲料中分别加入1ml上述稀释液，充分混匀，依次得到含杀虫蛋白浓度为0.25μg/g、0.5μg/g、1μg/g、2μg/g、4μg/g、8μg/g、16μg/g、32μg/g的饲料，分装于经消毒(5％福尔马林浸泡)的24孔细胞培养板中。用毛笔轻轻接入棉铃虫1-2龄幼虫，每孔一头，每处理重复三次，放置25℃光照培养箱中，培养七天调查试虫的体重。
Cry2Ah-likesp蛋白对棉铃虫的室内杀虫活性测定操作步骤同上。
以清水为阴性对照。
蛋白对试虫的IC50具体结果见表2。
2.抗性品系棉铃虫的室内杀虫活性测定
根据实施例2中的杀虫蛋白的定量结果，首先将杀虫蛋白进行系列稀释；然后称取8份10g人工饲料中分别置于灭菌培养皿中，并向每份的人工饲料中分别加入1ml上述稀释液，充分混匀，依次得到含杀虫蛋白浓度为2μg/g、4μg/g、8μg/g、16μg/g、32μg/g、64μg/g的饲料，分装于经消毒(5％福尔马林浸泡)的24孔细胞培养板中。用毛笔轻轻接入棉铃虫1-2龄幼虫，每孔一头，每处理重复三次，放置25℃光照培养箱中，培养七天调查试虫的体重。
Cry2Ah-likesp蛋白对棉铃虫的室内杀虫活性测定操作步骤同上。
以清水为阴性对照。
蛋白对试虫的IC50具体结果见表2。
3.敏感品系亚洲玉米螟(O.furnacalis)的室内杀虫活性测定
配置饲料，50g干饲料加入48g超纯水得到湿饲料。
根据杀虫蛋白的定量结果，首先将杀虫蛋白进行系列稀释。
称取6份2g湿饲料于每个无菌的培养皿中，并向每份的人工饲料中分别加入200μl上述稀释液，充分混匀。依次得到含杀虫蛋白浓度为2μg/g、4μg/g、8μg/g、16μg/g、32μg/g、64μg/g的饲料。每个培养皿接20头初孵幼虫，每个处理重复3次，26℃生化培养箱中保温，培养7天后调查试虫的体重。
Cry2Ah-likesp蛋白对亚洲玉米螟的室内杀虫活性测定操作步骤同上。
以清水为阴性对照。
蛋白对试虫的IC50具体结果见表2
表2
由表2的结果可知，本申请的Cry2A-like杀虫蛋白对棉铃虫和亚洲玉米螟的活性均显著地优于已知的人工突变杀虫蛋白Cry2Ah-likesp。将本申请的Cry2A-like杀虫蛋白用于防治棉铃虫和/或亚洲玉米螟可以显著地降低生产成本，从而带来更为可观的经济效益。
虽然本发明已经参照其优选地具体实施方式进行了描述，但是本领域的技术人员应该理解在没有脱离本发明的真正的精神和范围的情况下，可以进行的各种改变。例如，可以对本发明的主体、精神和范围进行多种改变以适应特定的情形、材料、材料组合物或方法步骤。所有的这些改变均包括在本发明的权利要求的范围内。并且利用上述揭示的技术内容做出些许的变动或修饰均等同于等效实施案例，均属于技术方案范围内。
LHA1760078 核 苷 酸 和 氨 基 酸 序 列 表
<110> 中国农业科学院植物保护研究所
<120> 一种新的杀虫蛋白及其核苷酸序列
<130> LHA1760078
<160> 8
<170> PatentIn version 3.5
<210> 1
<211> 1899
<212> DNA
<213>苏云金芽胞杆菌（Bacillus thuringiensis）
<223> cry2-like
<400> 1
1 ATG AAT AAT GTA TTG AAT AGC GGA AGA GCT
31 ACT AAT GGT GAT GCG TAT AAT GTA GTG GCT
61 CAT GAT CCA TTT AGT TTT CAA CAT AAA TCA
91 TTA GAT ACC ATA CAG GAA GAA TGG ATG GAG
121 TGG AAA AAA GAT AAT CAT ATT TTA TAT GTA
151 GAT CCT ATT GTA GGA ACT GTG GCT AGC TTT
181 CTT TTA AAG AAA GTG GGG AGT CTT GTA GAA
211 AAA AGA ATA TTA AGT GAG TTA CGG AAT TTA
241 ATA TTT CCT AGT GGC AGT ACT AAT CTA ATG
271 CAG GAT ATT TTA AGA GAG ACT GAA AAA TTC
301 CTG AAT CAG AGA CTT AAT ACA GAC ACT CTT
331 GCC CGT GTA AAT GCG GAA TTG ACA GGG CTG
361 CAA GCA AAT GTA GAA GAG TTT AAT CGA CAA
391 GTA GAT AAT TTT TTG AAT CCT AAT CGA AAT
421 GCT GTA CCT TTA TCA ATA ACT TCT TCA GTA
451 AAT ACT ATG CAA CAA TTA TTT CTA AAT AGA
481 TTA CCC CAG TTT CAG ATG CAA GGA TAT CAA
511 TTG TTA TTA TTA CCT TTA TTT GCA CAA GCA
541 GCC AAT TTA CAT CTT TCT TTT ATT AGA GAT
571 GTT ATT CTT AAT GCA GAT GAA TGG GGA ATT
601 TCA GCA GCA ACA TTA CGT ACG TAT CAA AAT
631 CAC CTG AGA AAT TAT ACA AGA GAG TAT TCT
661 AAT TAT TGT ATA ACA ACG TAT CAA ACA GCG
691 TTT AGA GGT TTA AAC ACC CGT TTA CAC GAT
721 ATG TTA GAA TTT AGA ACA TAT ATG TTT TTA
751 AAC GTA TTT GAA TAT GTT TCT ATC TGG TCG
781 TTG TTT AAA TAT CAA AGC CTT CTA GTT TCT
811 TCT GGC GCT AAC TTA TAT GCA AGT GGT AGT
841 GGA CCA CAA CAG ACC CAA TCA TTT ACA TCA
871 CAA GAC TGG CCA TTT TTA TAT TCT CTT TTC
901 CAA GTT AAC TCA AAT TAT GTG TTA AAT GGC
931 TTT AGT GGC GCT AGA CTT ACG CAG ACT TTC
961 CCT AAT ATT GTT GGT TTA CCT GGT ACT ACT
991 ACA ACT CAC GCA TTG CTT GCT GCA AGG GTC
1021 AAT TAC AGT GGA GGA GTT TCG TCT GGT GAT
1051 ATA GGC GCT TCT CCT TTT AAT CAA AAT TTT
1081 AGT TGT AGT ACA TTT CTC CCA CCT TTG TTA
1111 ACA CCA TTT GTT AGA AGT TGG CTA GAT TCA
1141 GGT TCA GAT CGG GGG GGG ATT AAT ACC GTT
1171 ACC AAT TGG CAA ACA GAA TCC TTT GAG ACA
1201 ACT TTA GGT TTA AGG AGT GGT GCT TTT ACA
1231 GCT CGA GGT AAT TCA AAC TAC TTC CCA GAT
1261 TAC TTT ATC CGT AAT ATT TCT GGA GTT CCT
1291 TTA GTT GTT AGA AAT GAA GAT TTA AGA AGA
1321 CCG TTA CAC TAC AAT CAA ATA AGA AAT ATA
1351 GAA AGT CCT TCA GGA ACA CCT GGT GGA TTA
1381 CGA GCT TAT ATG GTT TCT GTG CAT AAC AGA
1411 AAA AAT AAT ATC TAT GCC GTT CAT GAA AAT
1441 GGT ACT ATG ATA CAT TTA GCG CCG GAA GAT
1471 TAT ACA GGA TTT ACT ATA TCG CCG ATA CAT
1501 GCA ACT CAA GTG AAT AAT CAA ACG CGA ACA
1531 TTT ATT TCT GAA AAA TTT GGT AAT CAA GGT
1561 GAT TCC TTA AGA TTT GAA CAA AGC AAT ACG
1591 ACA GCT CGT TAT ACC CTT AGA GGG AAT GGA
1621 AAT AGT TAC AAT CTT TAC TTA AGA GTA TCT
1651 TCA ATA GGA AAT TCC ACT ATT CGA GTT ACT
1681 ATA AAC GGT AGA GTT TAT ACT GCT TCA AAT
1711 GTT AAT ACT ACT ACA AAT AAC GAT GGA GTT
1741 AAT GAT AAT GGA GCT CGT TTT TCA GAT ATT
1771 AAT ATC GGT AAT GTA GTA GCA AGT GAT AAT
1801 ACT AAT GTA CCG TTA GAT ATA AAT GTG ACA
1831 TTA AAT TCG GGT ACT CAA TTT GAG CTT ATG
1861 AAT ATT ATG TTT GTT CCA ACT AAT CTT CCA
1891 CCA CTT TAT；
<210> 2
<211> 633
<212> PRT
<213>苏云金芽胞杆菌（Bacillus thuringiensis）
<223> Cry2-like
<400> 2
MNNVLNSGRA TNGDAYNVVA HDPFSFQHKS LDTIQEEWME WKKDNHILYV DPIVGTVASFLLKKVGSLVE KRILSELRNL IFPSGSTNLM QDILRETEKF LNQRLNTDTL ARVNAELTGL QANVEEFNRQVDNFLNPNRN AVPLSITSSV NTMQQLFLNR LPQFQMQGYQ LLLLPLFAQA ANLHLSFIRD VILNADEWGISAATLRTYQN HLRNYTREYS NYCITTYQTA FRGLNTRLHD MLEFRTYMFL NVFEYVSIWS LFKYQSLLVSSGANLYASGS GPQQTQSFTS QDWPFLYSLF QVNSNYVLNG FSGARLTQTF PNIVGLPGTT TTHALLAARVNYSGGVSSGD IGASPFNQNF SCSTFLPPLL TPFVRSWLDS GSDRGGINTV TNWQTESFET TLGLRSGAFTARGNSNYFPD YFIRNISGVP LVVRNEDLRR PLHYNQIRNI ESPSGTPGGL RAYMVSVHNR KNNIYAVHENGTMIHLAPED YTGFTISPIH ATQVNNQTRT FISEKFGNQG DSLRFEQSNT TARYTLRGNG NSYNLYLRVSSIGNSTIRVT INGRVYTASN VNTTTNNDGV NDNGARFSDI NIGNVVASDN TNVPLDINVT LNSGTQFELMNIMFVPTNLP PLY；
<210>3
<211> 23
<212> DNA
<213> 人工序列
<223> S5un2
<400> 3
GGAAGAACTACTATTTGTGATGC；
<210> 4
<211> 21
<212> DNA
<213> 人工序列
<223> S3un2
<400> 4
AATAGTTTAATTACCGCGAGC；
<210> 5
<211> 36
<212> DNA
<213> 人工序列
<223> cry2F
<400> 5
CCGGAATTCgATGAATAATGTATTG；
<210> 6
<211> 36
<212> DNA
<213>人工序列
<223> cry2R
<400> 6
CCCAAGCTTATAAAGTGGTGGAAG；
<210> 7
<211> 1899
<212> DNA
<213>人工序列
<223> cry2Ah-likesp
<400> 7
atgaatagtg tattgaatag cggaagaact actatttgtg atgcgtataa tgtagcggct
catgatccat ttagttttca acacaaatca ttagataccg tacaaaagga atggacggag
tggaaaaaaa ataatcatag tttataccta gatcctattg ttggaactgt ggctagtttt
ctgttaaaga aagtggggag tcttgttgga aaaaggatac taagtgagtt acggaattta
atatttccta gtggtagtac aaatctaatg caagatattt taagagagac agaaaaattc
ctgaatcaaa gacttaatac agacactctt gcccgtgtaa atgcggaatt gacagggctg
caagcaaatg tagaagagtt taatcgacaa gtagataatt ttttgaaccc taaccgaaac
gctgttcctt tatcaataac ttcttcagtt aatacaatgc aacaattatt tctaaataga
ttaccccagt tccagatgca aggataccaa ctgttattat tacctttatt tgcacaggca
gccaatttac atctttcttt tattagagat gttattctaa atgcagatga atggggaatt
tcagcagcaa cattacgtac gtatcgagat tacttgaaaa attatacaag agattactct
aactattgta taaatacgta tcaaagtgcg tttaaaggtt taaacactcg tttacacgat
atgttagaat ttagaacata tatgttttta aatgtatttg agtatgtatc tatctggtcg
ttgtttaaat atcaaagcct tctagtatct tctggcgcta atttatatgc aagtggtagt
ggaccacagc agacccaatc atttacttca caagactggc catttttata ttctcttttc
caagttaatt caaattatgt gttaaatggc tttagtggcg ctagacttac gcagactttc
cctaatattg ttggtttacc tggtactact acaactcacg cattgcttgc tgcaagggtc
aattacagtg gaggagtttc gtctggtgat ataggcgctt ctccgtttaa tcaaaatttt
agttgtagta catttctccc acctttgtta acaccatttg ttagaagttg gctagattca
ggttcagatc ggggggggat taataccgtt accaattggc aaacagaatc ctttgagaca
actttaggtt taaggagtgg tgcttttaca gctcgaggta attcaaacta tttcccagat
tattttatcc gtaatatttc tggagttcct ttagttgtta gaaatgaaga tttaagaaga
ccgttacact ataatcaaat aagaaatata gaaagtcctt caggaacacc tggtggatta
cgagcttata tggtatctgt gcataacaga aaaaataata tctatgccgt tcatgaaaat
ggtactatga ttcatttagc gccggaagat tatacaggat ttactatatc gccgatacat
gcaactcaag tgaataatca aacgcgaaca tttatttctg aaaaatttgg aaatcaaggt
gattccttaa gatttgaaca aagcaacacg acagctcgtt atacccttag agggaatgga
aatagttaca atctttattt aagagtatct tcaataggaa attccactat tcgagttact
ataaacggta gagtttatac tgcttcaaat gttaatacta ctacaaataa cgatggagtt
aatgataatg gagctcgttt ttcagatatt aatatcggta atgtagtagc aagtgataat
actaatgtac cgttagatat aaatgtgaca ttaaattcgg gtactcaatt tgagcttatg
aatattatgt ttgttccaac taatatttca ccactttat；
<210> 8
<211> 633
<212> PRT
<213>人工序列
<223> Cry2Ah-likesp
<400> 8
Met Asn Ser Val Leu Asn Ser Gly Arg Thr Thr Ile Cys Asp Ala Tyr
Asn Val Ala Ala His Asp Pro Phe Ser Phe Gln His Lys Ser Leu Asp
Thr Val Gln Lys Glu Trp Thr Glu Trp Lys Lys Asn Asn His Ser Leu
Tyr Leu Asp Pro Ile Val Gly Thr Val Ala Ser Phe Leu Leu Lys Lys
Val Gly Ser Leu Val Gly Lys Arg Ile Leu Ser Glu Leu Arg Asn Leu
Ile Phe Pro Ser Gly Ser Thr Asn Leu Met Gln Asp Ile Leu Arg Glu
Thr Glu Lys Phe Leu Asn Gln Arg Leu Asn Thr Asp Thr Leu Ala Arg
Val Asn Ala Glu Leu Thr Gly Leu Gln Ala Asn Val Glu Glu Phe Asn
Arg Gln Val Asp Asn Phe Leu Asn Pro Asn Arg Asn Ala Val Pro Leu
Ser Ile Thr Ser Ser Val Asn Thr Met Gln Gln Leu Phe Leu Asn Arg
Leu Pro Gln Phe Gln Met Gln Gly Tyr Gln Leu Leu Leu Leu Pro Leu
Phe Ala Gln Ala Ala Asn Leu His Leu Ser Phe Ile Arg Asp Val Ile
Leu Asn Ala Asp Glu Trp Gly Ile Ser Ala Ala Thr Leu Arg Thr Tyr
Arg Asp Tyr Leu Lys Asn Tyr Thr Arg Asp Tyr Ser Asn Tyr Cys Ile
Asn Thr Tyr Gln Ser Ala Phe Lys Gly Leu Asn Thr Arg Leu His Asp
Met Leu Glu Phe Arg Thr Tyr Met Phe Leu Asn Val Phe Glu Tyr Val
Ser Ile Trp Ser Leu Phe Lys Tyr Gln Ser Leu Leu Val Ser Ser Gly
Ala Asn Leu Tyr Ala Ser Gly Ser Gly Pro Gln Gln Thr Gln Ser Phe
Thr Ser Gln Asp Trp Pro Phe Leu Tyr Ser Leu Phe Gln Val Asn Ser
Asn Tyr Val Leu Asn Gly Phe Ser Gly Ala Arg Leu Thr Gln Thr Phe
Pro Asn Ile Val Gly Leu Pro Gly Thr Thr Thr Thr His Ala Leu Leu
Ala Ala Arg Val Asn Tyr Ser Gly Gly Val Ser Ser Gly Asp Ile Gly
Ala Ser Pro Phe Asn Gln Asn Phe Ser Cys Ser Thr Phe Leu Pro Pro
Leu Leu Thr Pro Phe Val Arg Ser Trp Leu Asp Ser Gly Ser Asp Arg
Gly Gly Ile Asn Thr Val Thr Asn Trp Gln Thr Glu Ser Phe Glu Thr
Thr Leu Gly Leu Arg Ser Gly Ala Phe Thr Ala Arg Gly Asn Ser Asn
Tyr Phe Pro Asp Tyr Phe Ile Arg Asn Ile Ser Gly Val Pro Leu Val
Val Arg Asn Glu Asp Leu Arg Arg Pro Leu His Tyr Asn Gln Ile Arg
Asn Ile Glu Ser Pro Ser Gly Thr Pro Gly Gly Leu Arg Ala Tyr Met
Val Ser Val His Asn Arg Lys Asn Asn Ile Tyr Ala Val His Glu Asn
Gly Thr Met Ile His Leu Ala Pro Glu Asp Tyr Thr Gly Phe Thr Ile
Ser Pro Ile His Ala Thr Gln Val Asn Asn Gln Thr Arg Thr Phe Ile
Ser Glu Lys Phe Gly Asn Gln Gly Asp Ser Leu Arg Phe Glu Gln Ser
Asn Thr Thr Ala Arg Tyr Thr Leu Arg Gly Asn Gly Asn Ser Tyr Asn
Leu Tyr Leu Arg Val Ser Ser Ile Gly Asn Ser Thr Ile Arg Val Thr
Ile Asn Gly Arg Val Tyr Thr Ala Ser Asn Val Asn Thr Thr Thr Asn
Asn Asp Gly Val Asn Asp Asn Gly Ala Arg Phe Ser Asp Ile Asn Ile
Gly Asn Val Val Ala Ser Asp Asn Thr Asn Val Pro Leu Asp Ile Asn
Val Thr Leu Asn Ser Gly Thr Gln Phe Glu Leu Met Asn Ile Met Phe
Val Pro Thr Asn Ile Ser Pro Leu Tyr。
Claims (16)
1.一种杀虫蛋白，其氨基酸序列如SEQ ID No.2所示。
2.编码如权利要求1所述的杀虫蛋白的核酸。
3.根据权利要求2所述的核酸，其特征在于，所述核酸的序列如SEQ ID No.1所示。
4.一种组合物，其包括如权利要求1所述的杀虫蛋白。
5.根据权利要求4所述的组合物，其特征在于，编码所述杀虫蛋白的核酸的序列如SEQID No.1所示。
6.一种含有根据权利要求2或3所述的核酸，且能产生如权利要求1所述的杀虫蛋白的转基因微生物。
7.根据权利要求6所述的转基因微生物，其特征在于，所述转基因微生物为芽胞杆菌(Bacillus)、假单胞菌(Pseudomonas)、肠杆菌(Escherichia)和酵母(Saccharomyces)中的至少一种。
8.根据权利要求7所述的转基因微生物，其特征在于，所述芽胞杆菌为苏云金芽胞杆菌(Bacillus thuringiensis)、枯草芽胞杆菌(Bacillus subtilis)、萎缩芽胞杆菌(Bacillus atrophaeus)和蜡样芽胞杆菌(Bacillus cereus)中的至少一种；所述假单胞菌为荧光假单胞杆菌(Pseudomonas fluorescens)；所述肠杆菌为大肠杆菌(Escherichia coli)。
9.一种制备如权利要求1所述的杀虫蛋白的方法，其包括利用根据权利要求6至8中任意一项所述的转基因微生物来生产所述杀虫蛋白。
10.根据权利要求9所述的方法，其特征在于，所述方法还包括在利用所述转基因微生物生产所述杀虫蛋白后，纯化所述杀虫蛋白使所述杀虫蛋白的纯度达到80％以上。
11.根据权利要求10所述的方法，其特征在于，使所述杀虫蛋白的纯度达到90％以上。
12.根据权利要求11所述的方法，其特征在于，使所述杀虫蛋白的纯度达到99％以上。
13.根据权利要求1所述的杀虫蛋白，根据权利要求4或5所述的组合物，以及根据权利要求6至8中任意一项所述的转基因微生物中的至少一种在防治鳞翅目(Lepidoptera)中的至少一种害虫中的应用。
14.根据权利要求13所述的应用，其特征在于，所述应用为在防治夜蛾科(Noctuidae)和螟蛾科(Pyralidae)中的至少一种害虫中的应用。
15.根据权利要求14所述的应用，其特征在于，所述应用为在防治铃夜蛾属(Helicoverpa)和秆野螟属(Ostrinia)中的至少一种害虫中的应用。
16.根据权利要求15所述的应用，其特征在于，所述应用为在防治棉铃虫(Helicoverpa armigera Hubner)和/或玉米螟(Ostrinia nubilalis)中的应用。
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| CN108795975B (zh) * | 2018-07-04 | 2021-10-22 | 中国农业科学院油料作物研究所 | 野生大豆相关蛋白在提高植物抗虫性中的应用 |
| CN109776659B (zh) * | 2019-03-14 | 2021-01-29 | 中国农业科学院生物技术研究所 | cry2Ah-vp基因在抗黏虫中的应用 |
| CN116284280B (zh) * | 2023-05-23 | 2023-08-08 | 莱肯生物科技(海南)有限公司 | 一种抗虫蛋白及其制备方法和应用 |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007107302A2 (en) * | 2006-03-21 | 2007-09-27 | Bayer Bioscience N.V. | Novel genes encoding insecticidal proteins |
| CN104611260A (zh) * | 2015-01-09 | 2015-05-13 | 东北农业大学 | 苏云金芽胞杆菌LTS290、杀虫基因cry57Ab、表达蛋白及其应用 |
| CN104744574A (zh) * | 2015-04-16 | 2015-07-01 | 中国农业科学院植物保护研究所 | 对玉米螟有杀虫活性的Bt蛋白及其应用 |
| CN105349555A (zh) * | 2015-12-11 | 2016-02-24 | 吉林省农业科学院 | 一种人工合成的Bt抗虫基因FLIa及其制备方法和应用 |
| CN105367633A (zh) * | 2014-08-28 | 2016-03-02 | 四川农业大学 | 一种BT蛋白CRY2Ab32、其编码基因及应用 |
| CN105777880A (zh) * | 2016-04-11 | 2016-07-20 | 中国农业科学院植物保护研究所 | 杀虫晶体蛋白、核酸、杀虫晶体蛋白的制备方法及其应用 |
| CN106011013A (zh) * | 2016-06-24 | 2016-10-12 | 东北农业大学 | 苏云金芽胞杆菌3-1-a、杀虫基因cry8Ax表达蛋白及其应用 |
| CN106008681A (zh) * | 2016-07-09 | 2016-10-12 | 河北省农林科学院植物保护研究所 | 一种杀虫蛋白及其核苷酸序列和应用 |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2397549T3 (es) * | 2001-01-09 | 2013-03-07 | Bayer Cropscience Nv | Proteínas insecticidas de Bacillus thuringiensis |
| US7557186B2 (en) * | 2002-03-27 | 2009-07-07 | Council Of Scientific & Industrial Research | Chimeric CrylE δ endotoxin and methods of controlling insects |
| US9522937B2 (en) * | 2007-03-28 | 2016-12-20 | Syngenta Participations Ag | Insecticidal proteins |
| ES2532145T3 (es) * | 2009-04-17 | 2015-03-24 | Dow Agrosciences Llc | Toxinas Cry insecticidas DIG-3 |
-
2017
- 2017-03-06 CN CN201710127378.1A patent/CN106928329B/zh active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007107302A2 (en) * | 2006-03-21 | 2007-09-27 | Bayer Bioscience N.V. | Novel genes encoding insecticidal proteins |
| CN105367633A (zh) * | 2014-08-28 | 2016-03-02 | 四川农业大学 | 一种BT蛋白CRY2Ab32、其编码基因及应用 |
| CN104611260A (zh) * | 2015-01-09 | 2015-05-13 | 东北农业大学 | 苏云金芽胞杆菌LTS290、杀虫基因cry57Ab、表达蛋白及其应用 |
| CN104744574A (zh) * | 2015-04-16 | 2015-07-01 | 中国农业科学院植物保护研究所 | 对玉米螟有杀虫活性的Bt蛋白及其应用 |
| CN105349555A (zh) * | 2015-12-11 | 2016-02-24 | 吉林省农业科学院 | 一种人工合成的Bt抗虫基因FLIa及其制备方法和应用 |
| CN105777880A (zh) * | 2016-04-11 | 2016-07-20 | 中国农业科学院植物保护研究所 | 杀虫晶体蛋白、核酸、杀虫晶体蛋白的制备方法及其应用 |
| CN106011013A (zh) * | 2016-06-24 | 2016-10-12 | 东北农业大学 | 苏云金芽胞杆菌3-1-a、杀虫基因cry8Ax表达蛋白及其应用 |
| CN106008681A (zh) * | 2016-07-09 | 2016-10-12 | 河北省农林科学院植物保护研究所 | 一种杀虫蛋白及其核苷酸序列和应用 |
Non-Patent Citations (2)
| Title |
|---|
| Bacillus thuringiensis strain BTSC6H8 cry2-like protein gene, complete cds;Genbank;《GenBank》;20111230;第1-2页 * |
| PirB-Cry2Aa hybrid protein exhibits enhanced insecticidal activity against Spodoptera exigua larvae;Xiaofeng Hu et al.,;《Journal of Invertebrate Pathology》;20140528;第120卷;第40-42页 * |
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