TAT-Apoptin基因原核表达载体的构建及其表达鉴定

为得到TAT-Apoptin 融合蛋白，以pMD-19T-Apoptin 为模板PCR 扩增出TATApoptin 基因，构建重组载体pET-32a-TAT-Apoptin，并对TAT-Apoptin 蛋白进行表达。PCR 和测序表明，成功构建重组载体pET-32a-TAT-Apoptin，进一步对其进行蛋白表达，发现该蛋 白能与Apoptin 多抗发生特异性结合。以上结果证明，TAT-Apoptin 基因原核表达载体构建成 功，并能在Rosetta 中进行分泌性表达。

Construction and expression Identification of prokaryotic expression Vector of TAT-Apoptin Gene

In order to obtain TAT-Apoptin fusion protein, TAT-Apoptin gene was amplified by PCR with pMD-19T-Apoptin as template, the recombinant vector pET-32a-TAT-Apoptin, was constructed and the TAT-Apoptin protein was expressed. PCR and sequencing showed that the recombinant vector pET-32a-TAT-Apoptin, was successfully constructed to further express the protein, and it was found that the protein could specifically bind to Apoptin polyantibodies. The above results show that TATapoptin base The prokaryotic expression vector was successfully constructed and secreted in Rosetta.