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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

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【作者】 高婧一王越赵雨杰房凯张劲松

【Author】 GAO Jing-yi1,WANG Yue3,ZHAO Yu-jie2,FANG Kai2,ZHANG Jin-song1 (1. Department of Ophthalmology,The Fourth Affiliated Hospital,China Medical University,Shenyang 110005,China;2. Department of Bioinformatics,College of Basic Medical Sciences,China Medical University,Shenyang 110001,China;3. Department of Ophthalmology,The Fourth People’s Hospital of Shenyang, Shenyang 110031,China)

【机构】 中国医科大学附属第四医院眼科沈阳市第四人民医院眼科中国医科大学基础医学院生物信息学教研室

【摘要】 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。

【Abstract】 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.

  • 【文献出处】 中国医科大学学报 ,Journal of China Medical University , 编辑部邮箱 ,2010年03期
  • 【分类号】R392
  • 【被引频次】2
  • 【下载频次】195
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