苏云金芽胞杆菌Vip3AaAdAa嵌合蛋白的构建及其杀虫活性分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0929

生物技术通报 ›› 2019, Vol. 35 ›› Issue (4): 51-56.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0929

苏云金芽胞杆菌Vip3AaAdAa嵌合蛋白的构建及其杀虫活性分析

于爽^1,2, 李帅², 李海涛¹, 刘荣梅¹, 高继国¹

1. 东北农业大学生命科学学院,哈尔滨 150030;
2. 牡丹江师范学院生命科学与技术学院,牡丹江 157011

收稿日期:2018-10-27 出版日期:2019-04-26 发布日期:2019-05-05
作者简介:于爽,教授,博士研究生,研究方向：植物生物化学与分子生物学;E-mail：swxys@126.com
基金资助:
国家重点研发项目（2017YFD0201200）,黑龙江省自然科学基金项目（C2016025）,国家自然科学基金青年基金项目（31401812）

Creation of Chimeric Protein Vip3AaAdAa in Bacillus thuringiensis and Analysis of Its Insecticidal Activity

YU Shuang^1,2, LI Shuai², LI Hai-tao¹, LIU Rong-mei¹, GAO Ji-guo¹

1. Northeast Agricultural University,Harbin 150030;
2. Mudanjiang Normal University,Mudanjiang 157011

Received:2018-10-27 Published:2019-04-26 Online:2019-05-05

摘要/Abstract

摘要：

Vip3A 类杀虫蛋白是一类新型杀虫蛋白,在氨基酸序列上与研究较为深入的Cry蛋白没有同源性,且昆虫受体结合位点与Cry蛋白也没有竞争关系。利用同源重组技术针对对甜菜夜蛾具有高活性的 Vip3Aa39 与无活性的Vip3Ad开展研究,成功构建了三个vip3AaAdAa型嵌合基因,将其转入大肠杆菌中表达嵌合蛋白Vip3AaAdAa1、Vip3AaAdAa2、Vip3AaAdAa3,并将这三种嵌合蛋白对甜菜夜蛾进行了杀虫活性测定。研究表明,三种嵌合蛋白对甜菜夜蛾具有杀虫活性,Vip3AaAdAa3蛋白的LC₅₀值比Vip3Aa39增加了1.4倍,Vip3AaAdAa1蛋白的LC₅₀值比Vip3Aa39增加了2.7倍,嵌合蛋白Vip3AaAdAa1和Vip3AaAdAa3的杀虫活性均低于Vip3Aa39;Vip3AaAdAa2的杀虫活性与Vip3Aa39差异不显著。

关键词: 苏云金芽胞杆菌, 嵌合蛋白, 甜菜夜蛾, 杀虫活性

Abstract:

Vip3A-type insecticidal proteins are a new class of insecticidal proteins. There is no homology in amino acid sequences with well-studied protein Cry,and the insecticidal sites are not competitive with insect receptor binding sites of Cry proteins. Homologous recombinant technique was used to study the insecticidal activity of Vip3Aa39 and inactive Vip3Ad against Spodoptera exigua. Three vip3AaAdAa chimeric genes were successfully created and were transformed into Escherichia coli to express chimeric protein Vip3AaAda1,Vip3AaAda2,and Vip3AaAda3. The insecticidal activities of these 3 chimeric proteins against S. exigua were determined. The results showed that the 3 chimeric proteins presented high insecticidal activities against S. exigua,the LC₅₀ of Vip3AaAda3 protein was 1.4 times higher than that of Vip3Aa39,and the LC₅₀ of Vip3AaAda1 protein was 2.7 times higher than that of Vip3Aa39. The insecticidal activities of chimeric proteins Vip3AaAda1 and Vip3AaAda3 were lower than that of Vip3Aa39,and the insecticidal activity of Vip3AaAdAa2 was not significantly different from that of Vip3Aa39.

Key words: Bacillus thuringiensis, chimeric protein, Spodoptera exigua, insecticidal activity

于爽, 李帅, 李海涛, 刘荣梅, 高继国. 苏云金芽胞杆菌Vip3AaAdAa嵌合蛋白的构建及其杀虫活性分析[J]. 生物技术通报, 2019, 35(4): 51-56.

YU Shuang, LI Shuai, LI Hai-tao, LIU Rong-mei, GAO Ji-guo. Creation of Chimeric Protein Vip3AaAdAa in Bacillus thuringiensis and Analysis of Its Insecticidal Activity[J]. Biotechnology Bulletin, 2019, 35(4): 51-56.

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苏云金芽胞杆菌Vip3AaAdAa嵌合蛋白的构建及其杀虫活性分析

Creation of Chimeric Protein Vip3AaAdAa in Bacillus thuringiensis and Analysis of Its Insecticidal Activity

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