中文摘要：

目的：针对Ⅱ型单纯疱疹病毒（HSV-2）包膜糖蛋白gD以及包膜糖蛋白gB的CTL表位，构建重组真核表达质粒pVAX—gD—CTL。方法：根据GeneBank提供的HSV-2G株糖蛋白D基因（gD）序列设计PCR引物，以已构建好的pc—gD为模板，特异性扩增HSV-2gm片段，将其克隆于pVAX1载体中构建成质粒pVAX—gD，再将其双酶切后引入CTL片段构建重组真核表达质粒pVAX—gD—CTL。结果：将重组的pVAX—gD—CTL真核表达质粒转化大肠杆菌后，筛选阳性克隆进行酶切及测序鉴定。结论：初步构建HSV-2 pVAX—gD—CTL重组真核表达质粒。

英文摘要：

Objective：To create a new vaccine against HSV, we aimed at constructing a recombinant eukaryotic plasmids pVAX - gD - CTL based on HSV - 2 gB and gD protein coding - gene. Methods： HSV - 2 gD protein coding - gene was amplified from pc - gD by PCR and inserted to pVAX1 plasmid, together with gB protein coding - CTL epitope. The recombinant plasmid pVAX - gD - CTL was tested in mice by inoculation via intramuscular injection. CTL activity was detected by Lactic Dehydrogenase （LDH） assay. Results ： Success of constructing recombinant plasmids was confirmed by restriction enzyme and DNA sequencing. Protein coded by gD gene was detected in COS -7 cells. The CTL activity was increased. Conclusion：We successfully constructed recombinant eukaryotic plasmids. The specific cell -mediated immune response against the virus was e- licited when BALB/C mice were immunized with pVAX- gD -CTL DNA vaccine.