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

摘要

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.