Antigenic epitope analysis, expression and purification of truncated forms of herpes simplex virus type Ⅱ glycoprotein D in eucaryotic cell

摘要

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.