将组成型表达的玉米泛素启动子与豇豆胰蛋白酶抑制剂基因CpTI连接,插人根癌农杆菌双T-DNA质粒,构建一个T-DNA结构域含有抗潮霉素选择标记基因hyg;另一个T-DNA结构域含有抗虫基因的双T-DNA单子叶植物表达载体,用以转化农杆菌菌株,再通过共培养转化玉米胚性愈伤组织.通过潮霉素培养基抗性筛选,用特异PCR扩增和Southern杂交检测,从分化再生的T₀代植株中,鉴定出7个转化CpTI基因的阳性植株.目前,正结合进行田间分离纯合和DNA分子鉴定,培育去除选择标记基因的转基因抗虫玉米自交系。

Cowpea trypsin inhibitor gene(CpTl)，promoted by uhiquitin promoter of constitutive expression，was inserted into double T-DNA plasmid of Agrobasterium tumefastions Monocotyledonous expression vector of doubleT-DNA was constructed with selective marker gene hyg(hygromycin resistance) in one T-DNA structural domain andinsect resistant gene in another T-DNA domain. Agrobasterium strain was transformed with this vector and used tctransform embryonic callus of maize by CO-culture Seven CpTI transgenie plants were identified from differentiatedand regenerated To population through resistant selection on hygromycin medium and molecular detection of specificPCR amplification and Southern hybridization At present，inbred lines of transgenie insect resistant maize with se-lective marker gene removed are in separating and inbreeding program assisted by DNA marker detection.

国家自然科学基金项目(30070476)