农杆菌介导的苏云金杆菌抗虫基因cryIA(b)和cryIA(c)在水稻中的遗传转化及蛋白表达

通过农杆菌介导法用含有抗潮霉素和 Ｇ Ｕ Ｓ 基因的双元载体将杀虫结晶蛋白基因ｃｒｙ Ｉ Ａ（ ｂ） 和ｃｒｙ Ｉ Ａ（ｃ） 导入到籼、粳稻幼穗愈伤组织中，然后经过在含有不同浓度潮霉素的培养基上进行数次筛选，获得一批 Ｂｔ 转基因株。经 Ｐ Ｃ Ｒ、 Ｓｏｕｔｈｅｒｎ 杂交及 Ｗｅｓｔｅｒｎ 印迹分析证实此二基因已整合进水稻中，饲虫试验结果表明，转基因株具有１００ ％ 杀虫率。

Agrobacterium mediated Transformation of Insecticidal Bacillus thuringiensis cryIA(b) and cryIA(c) Genes and Their Expression in Rice

The insecticidal Bacillus thuringiensis (Bt) cryIA(b) and cryIA(c) genes carried in the T DNAs of binary vectors which contain hygromycin resistance gene and GUS gene,were used to transform the callus of rice young infloresences via Agrobacterium transformation.Both of the insecticidal genes were successfully integrated into the genome of japonica and indica rice.Through stringent selection of the infected calli on the medium added with hygromycin,the transgenic rice plants were regenerated from the hygromycinresistant calli.The results of molecular biological assays with PCR and Southern hybridization confirmed that the cryIA(b) and cryIA(c) genes were integrated into the genome of rice.Western blotting analysis for the transgenic plants showed that cryIA(b) and cryIA(c) gene expressed and produced Bt protein.100% mortality rate was observed when stemborers were fed on the stems or leave from the transgenic rice.