苏云金芽胞杆菌V4菌株cry杀虫基因的鉴定、表达及杀虫活性分析  

Identification,Expression and Insecticidal Activity Analysis of cry Genes from Bacillus thuringiensis V4

作　　者：何宝楠 李海涛[1] 刘荣梅[1] 王博[1] 高继国[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《生物技术通报》2014年第9期125-130,共6页Biotechnology Bulletin

基　　金：国家高技术研究发展计划资助项目(2011aa10a203);转基因生物新品种培育(国家科技重大专项)(2014ZX0800913B-002);国家基础科学人才培养基金资助项目(J1210069);东北农业大学博士启动基金项目(2010RCB54)

摘　　要：旨在给生物防治提供更多的基因资源,以从本实验室分离的300株苏云金芽胞杆菌基因组为模板,利用cry1类全长通用引物进行PCR cry1类基因鉴定。经过PCR-RFLP鉴定出菌株V4含有cry1Ea基因,进一步研究发现该菌株还含有cry2Aa基因,并成功克隆到了这两个新基因,被Bt国际命名委员会正式命名为cry1Ea12和cry2Aa16。将两种基因在大肠杆菌Rosetta(DE3)中表达,表达蛋白进行杀虫活性测定,结果表明两种蛋白杀虫活性不理想,但对试虫存在明显的体重抑制,而V4菌株蛋白具有较高的杀虫活性。In order to provide more gene resources for biological control, we used the cry1 class full-length primers and genomic DNA of 300 Bacillus thuringiensis strains which stored in our laboratory as the templates to identify cry1 gene-types by PCR. The strain V4 was identified containing a cry1 Ea gene by PCR-RFLP, further study found that the strain also contained cry2 Aa gene. The genes from the strain V4 were cloned, respectively. The two genes were designated as cry1Ea12 and cry2Aa16 by International Nomenclature Committee of Bacillus thuringiensis. Furthermore, the two genes were respectively transformed into Escherichia coli Rosetta（DE3）strain and the toxicity of the two genes was evaluated. The results showed that the insecticidal activity of the proteins is not very efficient, but the weights of the test insects are inhibited obviously, and the insecticidal activity of the protein of the strain V4 is much higher.

关 键 词：苏云金芽胞杆菌 杀虫晶体蛋白 cry1Ea12基因 cry2Aa16基因 生物活性 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]