韧皮部特异性启动子的克隆及含绿色荧光蛋白报告基因新植物表达载体的构建

Cloning of phloem specific promoter and construction of new plant expression vector containing green fluorescence protein reporter gene

用PCR方法扩增得到了长度为966 bp的笋瓜韧皮部蛋白2基因启动子片段,克隆入pUCm-T载体后,获得了新的重组质粒pUCm-PSP.分别用限制性内切酶酶切重组质粒和pCAMB IA1302植物表达载体,经回收、连接、转化和鉴定后,构建了由PSP驱动绿色荧光蛋白(GFP)报告基因的新型植物表达载体pHZ03.利用细胞感受态法将新植物表达载体分别导入根癌农杆菌EHA105、GV3101、LBA4404和发根农杆菌R i15834中.

A 966 bp fragment of phloem protein 2 （PP2） gene in the promoter region of Cucurbita maxima was amplified by PCR. The fragment was cloned into pUCm-T vector, and a new recombined vector named pUCm-PSP was obtained. A new plant expression vector named pHZ03 in which the reporter gene GFP is droved by PSP was constructed after cutting two vectors pUCm-PSP and pCAMBIA1302 with restriction enzymes, subsequently recovering, ligation, transformation and identification. The recombined plant expression vector was transferred into Agrobacteriurn tumefaciens strains of LBA4404, GV3101, EHA105 and A. rhizogenes strain of Ri15834 by using cell competent method.