Cry2A基因植物表达载体构建及其转化子生长曲线的建立

Construction of plant expression vector of Cry2A gene and establishing the growth curve of its transformat

根据定向克隆原则,以pCAMBIA2301为载体骨架,Cry2A基因为目的基因,构建了大小为15354 bp的植物表达载体p2AST2301。新载体带有抗虫基因Cry2A和筛选标记基因NPTⅡ,适合通过农杆菌进行遗传转化,并以抗生素G418为筛选底物。同时将新载体转化根癌农杆菌EHA105,获得了用于植物遗传转化的工程菌,并且明确了工程菌D600 nm值在1.0-1.6之间活力最强。

Plant expression vector p2AST2301,which has the target gene of Cry2A gene and the screening marker gene of NPTⅡ gene,was constructed.The size of p2AST2301 is 15354 bp,it is suitable for transformation and the transgenic plant should be selected by G418.The new vector was introduced into Agrobacterium EHA105,and when the D600 nm value ranged from 1.0 to 1.6,the engineering bacteria was in the logarithmic growth phase with the highest vitality.