【摘要】 首先从油菜叶绿体基因组克隆得到包含rps7基因在内的 1.0kbDNA片段和包含ndhB基因在内的 2 .4kbDNA片段 ,同时从苏云金芽孢杆菌质粒上克隆得到一个全长 3.5kb的Bt杀虫蛋白基因cry1Aa。然后以rps7和ndhB基因作为同源重组片段 ,成功构建了包含Bt杀虫蛋白基因和aadA抗壮观霉素基因的油菜叶绿体定点转化载体 ,并对克隆菌体总蛋白进行了生物杀虫试验。结果表明 ,Bt杀虫蛋白基因能够得到表达 ,并且对棉铃虫幼虫有很强的毒杀作用。含抗虫基因的油菜叶绿体转化载体的构建在国内外为首次报告 ,该载体对于油菜叶绿体基因组研究和培育高抗虫品种具有重要应用价值更多还原

【Abstract】 WT5BZ]1.0 kb and 2.4 kb DNA fragment containing rps7 gene and ndhB gene, respectively, were cloned from oilseed rape chloroplast genome. A full length 3.5 kb Bt toxin gene ( cry1Aa ) was also cloned from the plasmid of Bacillus thuringiensis. The vector containing Bt toxin gene and aadA gene for oilseed rape chloroplast transformation was constructed with the rps7 gene and ndhB gene as homologous recombinant fragments. Bioassays using crude expressed products from host strain of the vector showed that Bt toxin gene was expressed and its products were toxic to larvae of Heliothis armigera. The chloroplast transformation vector will be of great value to oilseed rape chloroplast transformation and to the insect resistant crop improvement.更多还原