Vip3A杀虫蛋白随机重组文库的构建与分析

生物技术通报 ›› 2013, Vol. 0 ›› Issue (3): 160-165.

Vip3A杀虫蛋白随机重组文库的构建与分析

张宁¹,刘荣梅¹,束长龙²,张杰²,李海涛¹,高继国¹

1. 东北农业大学生命科学学院，哈尔滨 150030 ；2. 中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室，北京 100193

收稿日期:2012-08-27 修回日期:2013-03-21 出版日期:2013-03-20 发布日期:2013-03-21
作者简介:张宁，女，硕士研究生，研究方向：生防微生物分子生物学；E-mail ：zhangning_13@hotmail.com
基金资助:
植物病虫害生物学国家重点实验室开放基金项目（SKL2011P03），黑龙江省教育厅科学技术研究项目计划（12521021）， “863”计划课题（2011AA10A203）

Construction and Analysis of Vip3A Insecticidal Protein Random Recombination Library

Zhang Ning¹, Liu Rongmei¹, Shu Changlong², Zhang Jie², Li Haitao¹, Gao Jiguo1

1. College of Life Science, Northeast Agricultural University, Harbin 150030 ；2. State Key Laboratory of Biology Chinese Academy of Agricultural Sciences Institute of Plant Protection, Plant Diseases and Insect Pests, Beijing 100193）

Received:2012-08-27 Revised:2013-03-21 Published:2013-03-20 Online:2013-03-21

摘要/Abstract

摘要： 营养期杀虫蛋白（Vegetative Insecticidal Proteins，VIPs）作为苏云金芽胞杆菌（Bacillus thuringiensis，Bt）一类新型杀虫蛋白，丰富了Bt 的杀虫谱，是一种用于植物保护的重要杀虫蛋白。为了提高VIPs 蛋白的杀虫活性，扩展其杀虫谱，利用 Gateway 技术对vip3Aa39 和vip3Aa11 基因进行同源重组突变，并克隆到入门载体pDONR221 中，构建了重组文库。结果显示，得到了42 个基因型不同的重组质粒，其中有22 个蛋白序列不同的突变体。

关键词: 苏云金芽胞杆菌, 营养期杀虫蛋白, Gateway, 技术, 重组文库

Abstract: VIPs（Vegetative Insecticidal Proteins）from Bacillus thuringiensis（Bt）are a new class of insecticidal proteins, which enrich the insecticidal spectrum of Bt, and which are important proteins for plant protection. In order to improve the insecticidal activity and expand the insecticidal spectrum of the VIPs, the homologous recombination of vip3Aa39 and vip3Aa11 were completed by Gateway technology, and the products were cloned into the entry vector pDONR221, constructing a mutant library. Result showed 42 recombinant plasmids of different genotypes and 22 mutants of different protein sequences were obtained. The construction and the analysis of the random mutation library makes it possible for getting the Vip proteins with high insecticidal activity, widely insecticidal spectrum and insecticidal specific mechanism research.

Key words: Bacillus thuringiensis, VIPs, Gateway technology, Recombination library

张宁,刘荣梅,束长龙,张杰,李海涛,高继国. Vip3A杀虫蛋白随机重组文库的构建与分析[J]. 生物技术通报, 2013, 0(3): 160-165.

Zhang Ning, Liu Rongmei, Shu Changlong, Zhang Jie, Li Haitao, Gao Jiguo. Construction and Analysis of Vip3A Insecticidal Protein Random Recombination Library[J]. Biotechnology Bulletin, 2013, 0(3): 160-165.

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