苏云金芽孢杆菌Cry1Ab和Cry1C同源重组及转化大豆

在研究和确定Cry1Ab和Cry1C蛋白对甜菜夜蛾和斜纹夜蛾协同杀虫作用的基础上,利用同源重组技术进行Cry1Ab和Cry1C之间的结构域交换获得高效融合蛋白,利用农杆菌介导的子叶节法将融合抗虫基因表达载体导入大豆中,从而获得抗虫转基因大豆新材料。采用PCR及蛋白试纸条检测法对获得的大豆再生植株进行鉴定,测得结果初世代阳性植株为121株。对部分后代转基因植株的遗传分析表明外源基因能够稳定遗传。室内接种斜纹夜蛾鉴定表明,含有重组基因的转基因大豆对斜纹夜蛾有较强的抗性。因此,利用同源重组技术进行Cry1Aa和Cry1C之间的结构域交换,是获得高抗食叶性害虫大豆的有效途径。

Homologous Recombination of Bacillus thuringiensis Cry1Ab and Cry1C and Transformation of Soybean

On the basis of the study and determination of the synergistic insecticidal effect of Cry1Ab and Cry1C protein on the beet armyworm and the Spodoptera litura,the high efficient fusion protein was obtained by the exchange of Cry1Ab and Cry1C by the homologous recombination technology,and the fusion gene expression vector was introduced into soybean by the Agrobacterium-mediated cotyledon node method.A new insect resistant transgenic soybean material was obtained.PCR and protein test strip method were used to identify the regenerated plants of soybean.The results showed that the positive plants in the first generation were 121 plants.Genetic analysis of the first generation transgenic plants showed that the foreign genes could be stable to the next generation.