单纯疱疹病毒2型gG-2"独特区"基因的原核表达及其血清学诊断价值

目的在大肠杆菌中表达HSV-2的gG-2"独特区"基因(gG-2T)并对其免疫学特性进行初步研究。方法克隆HSV-2的gG-2T,并在大肠杆菌中表达gG-2T融合蛋白。通过Western blot和间接ELISA鉴定融合蛋白的活性。结果通过原核系统表达的gG-2T融合蛋白可以与GST多克隆抗体结合并在46 ku附近出现特异性结合条带。并且能够与确诊的HSV-2患者血清反应,而不与HSV-1患者血清和正常人血清反应。结论获得了HSV-2 gG-2T融合蛋白,它可以与GST多克隆抗体特异性结合并且可以同HSV-2型患者血清特异性反应;获得的gG-2T蛋白为研究以gG-2蛋白作为靶区域的诊断试剂盒和疫苗研发打下实验基础。

Expression in prokaryotic system and serodiagnostic value of herpes simplex virus type 2 glycoprotein G-2 unique domain

Objective To express the unique gene of herpes simplex virus type 2(HSV-2) gG-2(gG-2T) in E.coli BL21,and study preliminarily the lemologic characteristics of gG-2T protein. Methods We cloned the gene of gG-2T and had gG-2T fusion protein expressed in E.coli BL21,and identified its activity by Western blot and indirect ELISA.Results Glycoprotein G-2T fusion protein could combine with GST polyclonal antibody through gG-2T fusion protein expressed in prokaryotic system and SDS-PAGE had a specific binding strap in 46 ku nearby.The gG-2T fusion protein could react with HSV-2 patients' serum,but not with HSV-1 patients' serum and r-Globulin.Conclusion We obtained gG-2T fusion protein,which can combine with GST polyclonal antibody and react with HSV-2 patients' serum specifically.It will provide an experimental basis for diagnostic kit and vaccine of gG-2 protein target area.