Vip3A杀虫蛋白随机重组文库的构建与分析

Construction and Analysis of Vip3A Insecticidal Protein Random Recombination Library

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。

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 vip3Aal I 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.