中国生物防治学报2018，Vol.34Issue(1)：79-85,7.DOI:10.16409/j.cnki.2095-039x.2018.01.009
苏云金芽胞杆菌Vip3Aa11蛋白C端点突变对其杀虫活性的影响
The Influence of C-terminus Site-directed Mutation on the Toxicity of Bacillus thuringiensis Vip3Aa11 Protein
摘要
Abstract
Vegetative insecticidal proteins (VIPs) of Bacillus thuringiensis show a wide insecticidal spectrum against a wide variety of lepidopteran pests.The insecticidal activity and specificity of the Vip3Aa protein derived from different Bt strains are highly different,although the similarities between these proteins are higher than 95％.To determine whether the residues from the Vip3Aa11 C-terminus contribute to the insecticidal activity,three mutants (S543N,D547E and T681V) of Vip3Aa11 were constructed by site directed mutagenesis.Through SDS-PAGE,the mutants produced a 88 kD fusion protein,indicating that all mutants had successful expression in E.coli.Bioassay detection indicated that the insecticidal activity of the S543N mutant against Spodoptera exigua increased obviously,and the LCs0 value was approximately 5-fold higher than that of the Vip3Aa11,while the activity of D547E mutant against S.exigua decreased obviously.The activities of all the mutants against Helicoverpa armigera had no obvious changes.The results indicated that the site directed mutagenesis of some amino acids from Vip3Aa11 C terminus had an effect on insecticidal activity,and the effect was not consistent against different Lepidopteran pests even in the same mutant.In this study,the differences of insecticidal activity between Vip3Aa11 and mutant proteins serve a guideline for the study of Vip3Aa protein structure and activity mechanism.关键词
苏云金芽胞杆菌/营养期杀虫蛋白/定点突变/活性分析Key words
Bacillus thuringiensis/vegetative insecticidal proteins/site directed mutation/activity分类
农业科技引用本文复制引用
雒国兴,刘荣梅,李海涛,张金波,高继国,张杰..苏云金芽胞杆菌Vip3Aa11蛋白C端点突变对其杀虫活性的影响[J].中国生物防治学报,2018,34(1):79-85,7.基金项目
国家自然科学基金(31401812) （31401812）
国家重点研发计划(2017YFD0201201) （2017YFD0201201）
黑龙江自然科学基金(C2016025) （C2016025）
植物病虫害生物学国家重点实验室开放基金(SKLOF201705) （SKLOF201705）
