抗旱、耐盐基因P5CS植物表达双元载体的构建

Construction of Drought and Salt Resistance Gene P5CS in Plant Expression Binary Vector

以含有P5CS的重组质粒pBIP5CS-F129A为模板,通过PCR的方法获得了目的基因P5CS(1905 bp)并将其克隆到pBS-T载体上。利用限制性核酸内切酶BamHⅠ和SalⅠ将P5CS基因从克隆载体上消化下来,定向插入到无GUS基因植物表达载体pCHF3的CaMV35S启动子和NOS终止子之间。通过冻融法将此表达载体导入农杆菌LBA4404中,经PCR鉴定表明,P5CS基因植物表达双元载体构建成功。

The gene P5CS（ 1 905 bp）was amplified via PCR using pBIP5CS-F129A plasmid as template and cloned into pBS-T vector.The BamH Ⅰ/Sal Ⅰ fragment from the recombinant plasmid pBS-T/P5CS was ligated directively between CaMV35S promotor and NOS terminater of the expression vector pCHF3 without GUS gene. The plant expression vector pCHF3/P5CS was transfered into Agrobacterium tumefaciems strain LBA4404 by freeze-thaw method,and the expression binary plasmid was further identified by PCR. The results showed that pCHF3/P5CS vector was constructed successfully.