乙醇酸氧化酶基因植物表达载体构建及转化苹果的研究

为了研究GO基因在植物中的功能,获得转基因植株,用限制性内切酶将乙醇酸氧化酶(GO)基因从克隆载体pBSSP5上切下,连接到植物双元表达载体pBin117的CaMV 35S启动子和NOS 终止子之间,成功构建了GO基因植物表达载体pBinGO.在此基础上以嘎拉苹果叶片为受体,通过根癌农杆菌介导法将GO基因导入苹果,获得了2个抗性芽,PCR检测初步表明GO基因已整合到嘎拉苹果基因组中.

Construction of GO Plant Expression Vector and Its Transformation in Apple

In order to study the gene function of GO,obtain the transgenic plant,a plant expression vector pBinGO was constructed by recombinating the Glylate oxidase gene fragment from cloning vector pBSSP5 and the binary vector pBin117 at the EcoR Ⅰ site.The fragment of Glylate oxidase gene was inserted between of CaMV 35S promotor and NOS terminator in pBin117.The identification results showed that the plant expression binary vector pBinGO was constructed successfully.Subequentely GO gene was introduced into apple leaves mediated by Agrobacterium tumerfaciens.Two transgenic resistant plants were proved by PCR preliminarily that the GO gene had integrated into Apples genome.