【摘要】 本文用HindⅢ/BamHⅠ对含Cry Ⅳ D基因的pGEMl质粒进行双酶切,获得了含CryⅣ和20-kDa蛋白质基因的5.4kb片段,将其与穿梭质粒pⅪ61连接,转化到感受态大肠杆菌TG1细胞,对克隆的重组质粒经质粒抽提,进行斑点杂交,Southern分析加以证实后,电转化到苏云金芽胞杆菌无晶体突变株Bti.IPS. 78/11细胞中,可形成不规则的六边形晶体。SDS-聚丙烯酰胺凝胶电泳表明,工程菌表达72-kDa杀虫晶体蛋白和20-kDa的蛋白质,对致倦库蚊有很高的毒力,其LC₅₀值为0.53ug/ml。更多还原

【Abstract】 The plasmid pGEM1, carrying CryIVD, CytA and 20-kDa protein genes, was extracted and digested with Hind Ⅲ /BamH I . A 5. 4 kb fragment containing Cry IVD and 20-kDa protein gene was puri-fied and ligated to an E. coli TGl. The recombinant plasmid was analyzed by restriction map and Southern blotting to determine the correct insert. Then the recombinant was transferred into Bacillus thuringiensis acrystalliferous mutant Bti. IPS. 78/11 by electroporation. The clones with strong expressiveness were obtained. The engineered strains express 72-kDa and 20-kDa proteins and form irregular hexagon paras-pore crystal. One strain bioassayed is very toxic to Culex fatigans larvae with LC50 of 0. 53 μg/ml.更多还原