摘要
目的构建人疱疹病毒8型(HHV-8)融合蛋白原核表达载体,为诊断HHV-8感染奠定基础。方法构建HHV-8融合蛋白原核表达载体转化大肠杆菌DH5α,诱导表达融合蛋白ORF59-ORF65-K8.1。聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹(Western blot)法鉴定融合蛋白。融合蛋白对无偿献血人员血清进行检测。结果 SDS-PAGE显示融合蛋白约24KD,与预期值一致;Western blot提示该融合蛋白可以和HHV-8阳性血清特异性结合。融合蛋白包被酶连免疫吸附实验(ELISA)板后与无偿献血人员血清反应检测HHV-8感染情况,检测结果与市售HHV-8ELISA试剂盒检测结果一致。结论本文构建的融合蛋白可作为HHV-8感染的检测抗原用来筛查献血人员HHV-8感染情况及普通人群HHV-8感染流行病学调查。
Objective To construct the prokaryotic expression plasmid of HHV-8fusion antigen for diagnosis of HHV-8infection.Methods The combined fragment ORF59,ORF65and K8.1by fusion PCR was integrated into pQE-80Land transfected into E.coli DH5α.Fusion protein was induced to express by IPTG.SDS-PAGE and Western blot were employed to detect the fusion protein.Fusion protein was used to detect serum of blood donors.Results The combined plasmid pQE-80L-ORF59-ORF65-K8.1 was constructed successfully after verifying by restriction enzyme digestion and sequencing.The fusion protein was about 24KD and could be specific combined with HHV-8positive serum.The fusion protein had the same result to detect HHV-8with the HHV-8ELISA kit.Conclusion Fusion protein we construct can be used as diagnosis antigen to detect HHV-8of blood donors and common people.
出处
《重庆医学》
CAS
CSCD
北大核心
2013年第23期2703-2705,2709,共4页
Chongqing medicine
