HSV-gB、gD糖蛋白重组串联抗原表位蛋白的表达及鉴定

Expression and Characterization of the Recombinant Epitopes of HSV-gB and HSV-gD Protein

目的制备HSV-gB、gD蛋白串联表位重组蛋白,为单纯疱疹病毒(HSV)疫苗的研制提供新型抗原蛋白以及新的抗原制备方法。方法应用生物信息学软件laser gene DNASTAR分析HSV1-gB、gD和HSV2-gB、gD蛋白的抗原表位。选取9个表位,设计并合成表位串联重组蛋白编码基因X,构建其原核细胞表达重组体,在大肠杆菌BL21(DE3)表达该重组表位蛋白X,Western blot法(抗His标签)鉴定重组蛋白。结果构建了HSV-gB、gD蛋白串联表位重组蛋白X的原核表达体,在BL21菌中表达了X蛋白,Western blot法(抗His标签)鉴定了重组蛋白X。结论建立了制备HSV-gB、gD蛋白重组抗原表位蛋白的方法,为HSV疫苗的研制提供了可能的新型抗原蛋白。

Objective To prepare the recombinant epitopes of HSV-gB and HSV-gD protein and provides a new antigen protein for the development of herpes simplex virus （HSV）vaccine. Methods The epitopes of HSV-gB and HSV-gD protein were analyzed by epitope prediction software. A novel gene named X which encoded 9 predicted epitopes of HSV-Gb and HSV-gD protein was designed and synthesized using chemical method. X gene was cloned into vector PET-28a （＋）,expressed in Escherichia coli BL21 （DE3）,and analyzed by Western blot. Results X gene was successfully designed and expressed in Escherichia coli BL21（DE3）. Western blot analysis showed that recombinant X protein,which was with His marker,can be detected by anti-His antibody. Conclusion In this study we establish a new method to express recombinant epitope protein,which may be a new protein for developing vaccine against HSV infection.