大白菜orf224基因植物表达载体的构建及遗传转化

为了创制新的大白菜雄性不育材料,双酶切重组质粒pMD18-T-CMS7311-orf224和表达载体pWR306后,将CMS7311-orf224基因与线性表达载体pWR306进行定向连接,构建了细胞质雄性不育线粒体基因CMS7311-orf224的植物表达载体pWR-CMS7311-orf224,通过酶切和PCR验证pWR-CMS7311-orf224载体构建正确.采用快速冻融法,将表达载体导入农杆菌EHA105,转化大白菜材料06J28,对转化植株的GUS、PCR和RT-PCR检测表明,获得了2个大白菜转基因植株.

Construction and Transformation of the Plant Expression Vector Carrying orf224 Gene of Chinese Cabbage

In order to create a new cytoplasmic male sterile line of Chinese cabbage,the recombinant plasmid and expression vector pWR306 from double-enzyme digestion were linked directionally with the segments of CMS7311-orf224 gene to construct the plant expression vector pWR-CMS7311-orf224,which was introduced into Agrobacterium strain EHA105 by means of rapid frozen thaw method.The results of PCR amplification and restriction digestion showed that the fragment of CMS7311-orf224 gene was introduced into pWR306 plasmid and the expression vector pWR-CMS7311-orf224 was transferred into Agrobacterium successfully.By GUS,PCR and RT-PCR preliminary testing,we obtained two 06J28 transgenic lines.