H5N1型流感病毒M1蛋白的基因克隆与表达

目的：克隆、表达A型H5N1流感病毒基质蛋白M1,为研制新的流感快速检测方法奠定基础。方法：应用PCR方法扩增M1的全基因序列,克隆至PET32a载体上,经测序分析确认后,转化感受态大肠杆菌BL21（DE23）,经IPTG诱导表达M1蛋白后,对其表达产物进行SDS-Page和Western Bloting分析。结果：SDS-Page电泳显示M1蛋白在BL21（DE23）大肠杆菌中成功表达,Western Bloting分析显示表达产物可以与M1多克隆抗体发生特异性反应。结论：成功克隆和表达了流感病毒H5N1的基质蛋白M1,为进一步制备高滴度单克隆抗体、研制新的流感检测方法奠定了基础。

Clone and expression of M1 protein of influenza A H5N1 virus

Objective：To clone and express the gene of matrix protein 1（M1） of influenza A virus,and build the basis of new fast examination method for influenza virus.Methods：The whole sequence of M1 gene was amplified by RT-PCR,and cloned into the expression vector PET32a.The gene in the recombinant plasmid was then transformed into E.coli BL21（DE3）,which was induced with 1 mmol/L IPTG.The whole protein of E.coli was analyzed by SDS-PAGE and western bloting.Results：M1 protein was expressed in E.coli BL21（DE23） and specially bound to the polyclonal antibody of M1.Conclusion：M1 protein of influenza A virus was successfully cloned and expressed,which provide basis for the preparation of high titer monoclonal antibody and manufacture of fast examination kit for influenza virus.