Antigenic epitope analysis, expression and purification of extracellular region fragment of herpes simplex virus type I glycoprotein B in eukaryotic cell

摘要

In order to prevent infection from herpes simplex virus type I (HSV-1), and make the found for the construction of subunit vaccine, a truncated (extracellular region gene fragment) HSV-1 gB gene was cloned and expressed in eucaryotic cells by genetic engineering technology. After analizing the amino acid sequences of gB1 protein using Anthewin software, its antigenic epitopes were screened. Then the eucaryotic expression vector pCEP4 was constructed that contained the chemosynthetic selected sequence encoding HSV-1 gB with optimized signal peptide. Subsequently, transfecting the recombiant plasmid into HEK 293 cells and selecting it with hygromycin B, gB1 recombinant protein was then expressed in culture medium and purified through Ni affinity chromatography. The recombinant eukaryotic plasmid pCEP4-gB1 was constructed successfully, which was confirmed by the following SDS-PAGE and Western blot analysis. One major protein band (approximate molecular weights of 75 kDa) was observed, while no corresponding band was detected in uninduced extract. ELISA detection indicated that expressed gB1 had good antigenicity. The construction of truncated HSV-1 subunit vaccine is identified; it is powerful to induce immunoreactions and is promising for the treatment of HSV-1.