cry2Aa9m抗虫基因植物表达载体构建及对大豆的遗传转化

Construction of plant expression vector of insect-resistant gene cry2Aa9m and transformation into Glycine max L.Merr

cry2Aa9m基因编码的杀虫晶体蛋白(ICPs)对鳞翅目(Lepidoptera)和双翅目(Dipter)昆虫具有显著的毒杀作用,可防治大豆食心虫等害虫。试验构建了由豆荚特异性启动子Pmsg调控的cry2Aa9m基因的植物表达载体pCMB2A,以抗除草剂bar基因为筛选标记,通过农杆菌介导法对绥农28大豆子叶节进行遗传转化,获得抗性株系8株。经PCR和RT-PCR检测结果证明,cry2Aa9m基因已经整合到大豆基因组中并得以表达。研究为抗大豆食心虫转基因育种产业化奠定基础。

The cry2Aa9m gene of Bacillus thuringiensis encode insecticidal crystal proteins which possess broad specificity exhibiting toxicity against lepidopteran as well as dipteran insect species, and can be used against soybean pod borer. The pod-specific expression vector pCMB2A was constructed. With in the vector, cry2Aa9m gene was driven by promoter Pmsg. Bar served as selection marker and insect resistant gene cry2Aa9m was transformed into Suinong28 via Agrobacterium-mediated method. Eight soybean transgenic resistant lines were acquired. PCR and RT-PCR detection indicated that the cry2Aa9m was intergrated into soybean genome and expressed by its promoter. There was useful for genetic engineering studies of resistance soybean bean pod borer.