甘蓝抗枯萎病基因FOC1植物表达载体构建及遗传转化

Construction of Plant Expression Vector and Transformation of Fusarium Wilt Resistance Gene FOC1 to Brassica oleracea

为研究甘蓝枯萎病抗性基因FOC1的抗性功能,利用前期克隆的FOC1基因,以pBI121质粒为植物表达载体,采用同源重组法构建FOC1基因的过表达载体;将构建好的重组质粒采用冻融法转入根癌农杆菌LBA4404菌株中,并通过农杆菌介导的甘蓝外植体转化法对感病甘蓝进行遗传转化,利用载体特异引物对获得的转基因植株进行PCR鉴定。结果表明,最终成功构建FOC1基因的过表达载体pBI121-35S-FOC1,并已成功整合到受体甘蓝基因组中。

In order to study the function of resistance gene of cabbage Fusarium wilt FOC1 in transgenic cabbage,the plant expression vector Pro35S：：FOC1/pBI121 was constructed by homologous recombination and transformed into Agrobacterium tumefaciens strain LBA4404 by freeze-thawing method.The genetic transformation of B.oleracea was conducted by Agrobacterium-mediated plant transformation method.The independent transformants were detected by PCR method.The results showed that the pBI121-35S-FOC1 expression vector had successfully integrated into the genome of acceptor cabbage.