单纯疱疹病毒Ⅱ型新型DNA疫苗的构建  

Construction of A New Type Herpes Simplex Virus Ⅱ DNA Vaccine

作　　者：关蕾[1] 赖维[1] 杨慧兰[2] 朱国兴[1] 许庆芳[1] 陆春[1] 

机构地区：[1]中山大学附属第三医院皮肤性病科,广东广州510630 [2]广州军区广州总医院皮肤性病科,广东广州510010

出　　处：《岭南皮肤性病科杂志》2009年第1期15-18,共4页Southern China Journal of Dermato-Venereology

基　　金：国家自然科学基金资助项目(项目编号:30371290)

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Gene Bank提供的HSV-2 G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-2 gD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。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.

关 键 词：HSV-2病毒 gD蛋白 CTL表位 DNA疫苗 

分 类 号：R392[医药卫生—免疫学]