甘薯IbNPR1基因表达载体的构建及转化烟草

为了研究甘薯NPR1基因在转基因植物广谱抗病中的应用,利用前期克隆的甘薯NPRI基因,以pCAMBIA1300质粒为基本载体,构建了由组成型CaMV35S启动子驱动IbNPR1基因的植物表达载体pCAMBIAl300+IbNPRl;然后,采用冻融法转入农杆菌EHA105菌株,通过叶盘法对烟草进行了遗传转化.经PCR检测,IbNPR1基因已整合到烟草基因组中.

Construction of Plant Expression Vector of IbNPR1 Gene and Transformation in Tobacco

In order to study the application of Ipomoea batatas NPRl gene in transgenic broad-spectrum resistant plants, the plant expression vector pCAMBIA1300+IbNPRl was constructed, in which the IbNPRl gene was driven by the constitutive promoter CaMV35S. The constructed expression vector was transformed into Agrobacterium tumefaciens EHA105 strain by freeze-thaw method. Leaf segments of tobacco were inoculated with Agrobacterium lumefaclens EHA105 containing plasmid pCAMBIA1300 +IbNPRl. The transgenic lines were selected by PCR detection, and the result showed that IbNPRl gene had been integrated into the genome of tobacco.