花生白藜芦醇合酶基因cDNA的克隆及植物表达载体的构建  

Cloning cDNA of Resveratrol Synthase and Construction of Its Plant Expression Vector

作　　者：刘蕾[1] 尉亚辉[1] 张儒[1] 荆西刚[1] 郭芝光[1] 朱剑光[1] 

机构地区：[1]西北大学生命科学学院陕西省生物技术重点实验室,陕西西安710069

出　　处：《生物技术》2005年第2期7-9,共3页Biotechnology

基　　金：陕西省自然科学基金资助项目(99JK122);陕西省教育厅专项科研资金(04JK133)

摘　　要：为得到含有白藜芦醇合酶cDNA(RS cDNA)并能表达白藜芦醇(Res)的转基因植物,以花生为材料,从其根部提取基因组DNA,并以其为模板,利用引物悬挂延伸PCR法扩增得到大小约1200bp的片段,将这个片段连接到克隆载体PBS-T上进行测序,测序结果证明,此片段就是花生的RS cDNA。将此片段再正向插入到植物表达载体PBI121的CaMV35s启动子和NOS终止子之间,对重组子进行筛选与鉴定,证明花生的RS cDNA已经正确插入PBI121中,成功的构建了植物表达载体PBI121L。Total DNA was extracted from roots of peanut(Arachis hypogaea).The cDNA of resveratrol synthase was cloned by over-hang extension PCR protocol using total DNA as template.The PCR products were cloned into PBS-T vector and named PBS-TL after its sequences was confirmed.Then the cDNA of resveratrol synthase was subcloned into plant expression vector PBI121 and named PBI121L.All these establish the foundation for getting the transgenic plant that contain RS cDNA and can express Res.

关 键 词：白藜芦醇合酶 白藜芦醇 引物悬挂延伸PCR法 重组子 植物表达载体 

分 类 号：Q785[生物学—分子生物学] Q782