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目的获得单纯疱疹病毒2型(HSV-2)糖蛋白G(gG2)免疫优势片段gG321-580并初步研究其抗原性。方法 PCR扩增gG321-580基因,利用Bac-to-Bac表达系统表达gG321-580His融合蛋白,Western blot和间接ELISA鉴定蛋白活性。结果在Sf9细胞中成功表达HSV-2 gG321-580His蛋白,gG321-580His能与小鼠抗HSV-2 gG2单抗结合,在60 KDa附近出现特异性结合条带,ELISA分析初步证实gG321-580His能够与HSV-2阳性血清反应,而不与HSV-1阳性血清和正常人血清反应。结论 gG321-580His蛋白获得成功表达,ELISA检测显示其具有良好的抗原性,为研究以gG321-580蛋白作为靶区域的HSV-2型特异性诊断试剂盒打下实验基础。
Objective To obtain the gG321-580 immunosuppressive fragment of the glycoprotein G (gG2) of herpes simplex virus type 2 (HSV-2) and study its antigenicity. Methods The gG321-580 gene was amplified by PCR. The gG321-580His fusion protein was expressed by Bac-to-Bac expression system. Western blot and indirect ELISA were used to identify the protein activity. Results The HSV-2 gG321-580His protein was successfully expressed in Sf9 cells, and gG321-580His could bind to mouse anti-HSV-2 gG2 McAb. The specific binding band appeared around 60 KDa. ELISA analysis confirmed that gG321-580His Reacts with HSV-2 positive serum, but not with HSV-1 positive serum and normal human serum. Conclusion The recombinant gG321-580His protein was successfully expressed and its antigenicity was demonstrated by ELISA. It laid the experimental foundation for the study of HSV-2 specific diagnostic kit with gG321-580 protein as target region.