无选择标记的甜瓜a-ACO1基因植物表达载体的构建及对烟草的共转化效果

Construction of the a-ACO1 gene of sweet melon expression vector with marker-free and its co-transformation to tobacco

从植物表达载体pCB-aACO1的T-DNA区段上切去nptⅡ基因,构建了无选择标记植物表达载体pCD-aACO1,其T-DNA区段只含有目的基因a-ACO1和GUS基因.从另一植物表达载体pCAMBIA2301的T-DNA区段切去GUS基因,构建了植物表达载体pCAMBIA2302,其T-DNA区段其只含有nptⅡ基因.用这2种新载体共转化烟草,在对40株抗性转基因烟草的PCR检测中,有14株扩增出a-ACO1基因,共转化率为35%.

Culturing marker-free transgenic plant can improve the security of transgenic plant,the key is to get the co-transformation plant. In the experiments,a npt Ⅱ gene was cut from the binary plant expression vector pCB-aACO1, so a new binary plant expression vector pCD-aACO1 was formed, which T- DNA segment only harboured a a-ACO1 gene and a GUS gene. At the same time,GUS gene was cut from the other binary plant expression vector pCAMBIA2301, so a new binary plant expression vector pCAM- BIA2302 was formed,which T--DNA segment harboured a npt ]1 gene only. Tobacoo leaves were co-trans- formed by the two new vectors. By PCR analysis,a-ACO1 gene were indentified in 14 resistant transgenic plants out of 40,the rate of co-transformation was 35%.