田旋花EPSPS基因表达载体的构建及拟南芥转化

Construction of Expression Vector of Convolvulus arvensis EPSPS and Transformation of Arabidopsis thaliana

根据田旋花(Convolvulus arvensis L.)EPSPS基因(Gen Bank登录号:EU698030)c DNA序列,设计含有酶切位点的特异性引物,以田旋花c DNA为模板,合成EPSPS基因并连接到含有35S启动子和GUS基因的p BI121载体上,成功构建植物超表达载体p BI-EPSPS。采用氯化钙冻融法将其转入根癌农杆菌(Agrobacterium tumefaciens)中,然后用根癌农杆菌介导法转化拟南芥(Arabidopsis thaliana),共获得15株卡那霉素抗性拟南芥苗。对其中的5株进行PCR和RT-PCR检测,结果表明EPSPS基因已整合到拟南芥的基因组中并可以表达。

Based on the EPSPS gene c DNA（ Gen Bank accession number： EU698030） from field bindweed（ Convolvulus arvensis L.）,specific primers containing enzyme digestion sites were designed. With the template of c DNA from field bindweed,an EPSPS gene was obtained and inserted into plasmid p BI121 that contains promoter 35 S and GUS gene. A plant super expressed vector of p BI-EPSPS was constructed successfully and transferred to Arabidopsis thaliana by the Agrobacterium tumefaciens mediated transformation system. Fifteen transgenic plants were finally obtained,and five of them were analyzed by PCR and RT-PCR. The EPSPS gene was integrated into the genome of Arabidopsis thaliana and could be expressed.