Antigenic epitope analysis, ezpression and purification of truncated forms of herpes simplez virus type I glycoprotein D in eucaryotic ezpression system
To identify the possihility that the truncated herpes simplex virus type I glycoprotein D (gD1) expressed in eukaryotic cells can be used as a subunit vaccine of herpes simplex virus. Firstly, antigenic epilopes of gD1 protein were screened by analyzing the amino acid sequences using Anthewin software. Then, the extracellular region fragment gene of gD1 with optimized signal peptide 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, finally, gD1 recombinant protein was expressed in culture medium and purified through Ni affinity chromatography. From the experiment, the recombinant plasmid pCEP4-gD1 harboring synthetic gene of gD1 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 gD1 protein, was observed. In addition, ELISA detection showed that expressed gD1 has good antigenicity. Based on the above result, the successfully cloning and expression of gD1 recombinant protein in eucaryotic expression cells provides a basis for developing HSV subunit vaccine.
herpes simplez virus gD1 protein eucaryotic cell ezpression antigenicity
Wang Zheng-Mao Li Lin Wang Xing-Sheng Guan Wen-Yan Pan Mingjie Li Yue-Xi
Department of Biochemistry and Molecular Biology, School of Preclinical Medicine, Nanjing Medical Un The East-China Institute for Medical Biotechniques, Nanjing, 21002, 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
25-28
2009-04-27（万方平台首次上网日期，不代表论文的发表时间）