用电激转化法构建杀虫防病荧光假单胞菌  

CONSTRUCTION OF GENETICALLY ENGINEERED STRAINS OF PSEUDOMONAS FLUORESCENS AGAINST PLANT PATHOGEN AND INSECT PESTS BY ELECTROPORATING TRANSFORMATION

作　　者：张光焰[1] 张杰[1] 彭于发[1] 赵建周[1] 陈彩层[1] 徐樱[1] 黄大[1] 

机构地区：[1]中国农业科学院植物保护研究所,中国农业科学院蔬菜花卉研究所

出　　处：《中国农业科学》1995年第4期8-13,共6页Scientia Agricultura Sinica

基　　金：国家自然科学基金;"863"高技术计划青年基金

摘　　要：以广寄主范围质粒ｐＵＣＰ１８和ｐＵＣＰ１９为载体，用电激转化法，将苏云金杆菌杀虫蛋白基因ｃｒｙｌＡ（ｃ）片段导入了荧光假单胞菌Ｐ３０３菌株。Ｓｏｕｔｈｅｒｎ印迹分析和Ｗｅｓｔｅｒｎ印迹分析分别证实ｃｒｙｌＡ（ｃ）基因的导入和晶体蛋白在Ｐ３０３中的表达。生物活性测定结果表明，新构建的荧光假单胞工程菌株不仅保持了野生型自然菌株对小麦全蚀病有良好的抑菌活性，而且表现出对小菜蛾、玉米螟有显著的毒杀作用。In this study,an insecticidal crystal protein gene cryIA(c)of Bacillus thuringiensis via the broad host range plasmids pUCP18 and pUCP19 was transformed into biocontrol bacterium Pseudomonas fluorescens P303 by electroporation.southern blotting and Western blotting assays respectively verified the existence of cryIA(c)gene and expression of Bt toxin protein in the transformation.Results of bioassay showed that these new genetically engineered strains were not only inhibitory to the plant Pathogen Geumannomyces graminis var tritici, but also insecticidal to the corn borer and diamond back moth.

关 键 词：苏云金杆菌 荧光假单胞菌 电激转化 农药 

分 类 号：S482.7[农业科学—农药学]