Antigenic epitope analysis, ezpression and purification of truncated forms of herpes simplez virus type Ⅱ glycoprotein D in eucaryotic cell
(0) To express truncated herpes simplex virus type II glycoprotein D (gD2) in eucaryotic cells using genetic engineering technology. Antigenic epitopes of gD2 protein were screened by analizing the amino acid seqences using Anthewin software. The extracellular region fragment gene of gD2 with optimized signal peplide was synthesized chemically and cloned into eucaryotic expression vector pCEP4. After transfection of HEK 293 cells with the recombinant plasmid and selection with hygromycin B, gD2 recombinant protein was expressed in culture medium and purified through Ni affinity chromatography. The recombinant plasmid pCEP4-gD2 harboring synthetic gene of gD2 epitope mass region was constructed successfully. The expressed protein was confirmed with SDS-PAGE and Western blot analysis. And one major protein band, with approximate molecular weights of 46 kDa corresponding to the truncated forms of gD2 protein, was observed. ELISA detection showed that expressed gD2 has good antigenicity. The successfully cloning and expression of gD2 recombinant protein in eucaryotic expression cells provides a basis for developing HSV2 subunit vaccine.
herpes simplez virus gD2 protein eucaryotic cell ezpression antigenicity
Wang Zheng-Mao Li Lin Wang Xing-Sheng Guan Wen-Yan Pan Ming-Jie Li Yue-Xi
Department of Biochemistry and Molecular Biology, School of Preclinical Medicine, Nanjing Medical Un The East-China Institute for Medical Biotechniques, Nanjing, 210002, China Department of Biochemistry and Molecular Biology, School of Preclinical Medicine, Nanjing Medical Un
国际会议
International Symposium on Medical and Pharmaceutical Biotechnology(医药生物技术国际研讨会)
南京
英文
21-24
2009-04-27（万方平台首次上网日期，不代表论文的发表时间）