农杆菌介导法将Bt杀虫蛋白基因导入玉米自交系的研究

Study on Transferring Insecticide Protein Gene from Bacillus Thuringensis into Maize Callus Via Agrobacterium Mediation

利用β 葡萄糖苷酸酶(gus)基因的瞬时表达和稳定表达对农杆菌转化玉米愈伤组织的条件进行优化,通过用带有Bt抗虫基因和bar抗除草剂基因的根癌农杆菌LBA4404(ACT)转化玉米自交系7922、H99和P2的胚性愈伤组织,获得了抗性愈伤组织,并分化出植株。PCR检测初步证明Bt杀虫蛋白基因已整合到玉米再生植株中。

Factors influencing maize callus transformation mediated by Agrobacterium tumefaciens have been studied through gus transient expression and stable expression, such as different pre-cultivation time, different concentrations of AS and AgNO_3, different Agrobacterium growth phases, different infection time and infection media, et al. Maize embryogenic calli of inbred line 7922, H99 and P2 were transformed by Agrobacterium tumefaciens LBA4404(ACT). It was proved by PCR analysis that the target Bt. gene had been integrated into the genome of regenerated plants.