苏云金芽胞杆菌cry2Ab34基因的克隆、表达和杀虫活性分析  

Cloning,Expression,and Insecticidal Activities of Gene cry2Ab34 from Bacillus thuringiensis

作　　者：张月[1] 李海涛[1] 刘荣梅[1] 高继国[1] ZHANG Yue LI Hai-tao LIU Rong-mei GAO Ji-guo(College of Life Science, Northeast Agricultural University, Harbin 150030)

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》2017年第4期185-190,共6页Biotechnology Bulletin

基　　金：转基因生物新品种培育科技重大专项(2014ZX0800913B-002);国家基础科学人才培养基金资助项目(J1210069);国家自然科学基金项目(31401812);黑龙江省教育厅科学技术研究项目(12521021)

摘　　要：旨在对苏云金芽胞杆菌cry2Ab34基因进行分子鉴定和生物活测定,从杀虫活性高的Bt BJH500菌株中提取基因组DNA,通过PCR法克隆得到cry2Ab34基因(Gen Bank登录号为KX357382)。这个基因序列已被国际Bt基因命名委员会正式命名为cry2Ab34,结果显示,对cry2Ab34基因进行表达,获得大小约70 k D的表达产物,将该基因在大肠杆菌E.coli BL21中表达之后进行生物活性测定,结果表明对鳞翅目害虫小菜蛾(Plutella xyllostella)和棉铃虫(Helicoverpa armigera)都具有一定的杀虫活性,其校正死亡率10μg/m L为31.25%,100μg/m L为62.5%和校正死亡率10μg/m L为29.4%,100μg/m L为52.9%。In order to identify and detect insecticidal activities from Bacillus thuringiensis strain, the genomic DNA was extracted from the strain Bt BJH500 with high insecticidal activity, from which the insecticidal gene cry2Ab34 （ GenBank accession number ： KX357382 ） was cloned by PCR. The gene was designated as cry2Ab34 by International Gene Nomenclature Committee of Bt. Gene cry2Ab34 was expressed and the weight of expressed product was approximate 70 kD. The gene was expressed in Escherichia coli BL21 for the determination of biological activity, results showed that this gene presented certain insecticidal activity to Plutella xyllostella and Helicoverpa armigera of Lepidoptera peats, the corrected mortality of 10 μg/mL was 31.25%, of 100 μg/mL was 62.5 %, of 10 μg/mL was 29.4 %, and of 100 μg/mL was 52.9 %.

关 键 词：苏云金芽胞杆菌 cry2Ab34基因 杀虫晶体蛋白 杀虫活性 小菜蛾 棉铃虫 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]