HBsAg真核表达质粒的构建和鉴定

Construction of an Eukaryotic Expression Vector Inserted HBsAg Gene

目的 构建HBsAg真核表达质粒 .方法 用PCR的方法从质粒pEcob6中扩增出HBsAg基因 ,并定向克隆到真核表达质粒pcDNA3 .1(+) ,构建成重组质粒pcDNA3 .1-S ;然后用限制性内切酶消化和DNA序列测定鉴定 .结果 经酶切和DNA序列测定鉴定 ,证实重组质粒构建正确 .结论 真核表达质粒pcDNA3 .

Objective To construct a recombinant eukaryotic expression plasmid inserted HBsAg gene.Methods The full length gene of HBsAg from plasmid pEcob6 was amplified by PCR. Then the PCR product was digested with HindⅢ and BamHⅠ.Finally, the PCR product was cloned into plasmid pcDNA3.1(+).The accuracy of pcDNA3.1-S was confirmed by restriction enzyme digestion and DNA sequencing.Results Restriction enzyme digestion and DNA sequencing confirmed that the recombinant eukaryotic expression plasmid inserted HBsAg gene (pcDNA3.1-S) had been constructed correctly.Conclusions An eukaryotic expression plasmid pcDNA3.1- S has been constructed successfully.