苏云金芽胞杆菌CY1菌株的cry基因分析  

Characterization of cry Gene from a Novel Bacillus thuringiensis CY1

作　　者：吴会贤[1] 宋萍[1] 王勤英[1] 苏旭东[2] 

机构地区：[1]河北农业大学植物保护学院河北省农业病虫害生物防治工程技术研究中心,河北保定071001 [2]河北农业大学食品科技学院,河北保定071001

出　　处：《华北农学报》2007年第4期180-183,共4页Acta Agriculturae Boreali-Sinica

基　　金：河北省自然科学基金项目(303213);保定市科技局项目(05S001);河北省农业大学博士基金项目

摘　　要：通过醋酸钠-抗生素筛选法从河北省土壤中筛选得到了苏云金芽孢杆菌新菌株CY1。镜检可观察到伴孢晶体呈小菱形。SDS-PAGE分析结果表明菌株CY1表达的晶体蛋白分子量为130 kDa。利用25对通用引物检测cry1,cry1I,cry7,cry2,cry3,cry4/cry10,cry5,cry6,cry7,cry8,cry9,cry11,cry13/14,cry16/17,cry18,cry19,cry21,cry22,cry24/25,cry26/28,cry27/29,cry30,cry32,cry34/35,cry40基因,仅从CY1菌株中检测出cry7基因,该cry7基因N末端的1 215 bp片段所编码的氨基酸序列与已发表的Cry7Ab1(Acc No.:U04367)序列同源性达99%,仅有2个氨基酸的差异。用3对cry7Ab特异引物检测,进一步证实该菌株携带有cry7Ab基因。A novel strain CY1 of Bacillus thuringiensis（Bt） was isolated from the soil of Hebei province by Sodium acetate-antibiotic method. Bipyramidal inclusions could be visualized by microscope.SDS-PAGE analysis indicated that the strain producing a 130 kDa protein. The cry gene analysis of the strain was based on multiplex PCR with novel general primers that could detect the cry1 , cry11, cry7, cry2, cry3, cry4 /cry10, cry5, cry6, cry7, cry8, cry9, cry11 , cry13 /14, cry16/17, cry18, cry19, cry21, cry22, cry24/25, cry26/28, cry27/29, cry30, cry32, cry34/35 and cry40 genes. Only cry7 gene were found. PCR product of cry7 gene was sequenced. The cry7 gene fragments had 99 % homology to cry7Abl. The strains were further characterized to carry cry7Ab gene by additional PCRs with specific cry7Ab primers.

关 键 词：苏云金芽胞杆菌 CY1菌株 cry7基因 

分 类 号：S432[农业科学—植物病理学]