Retroviral Vector Mediated HCV E1E2 Envelope Protein Expression in Eukaryotic Cells and Animal Immune Experiment
Abstract: The recombinant retroviral vector pBABE-puro-E1E2 was constructed by inserting the full-length HCV e1e2 gene of H77 strain into pBABE-puro. Both the recombinant retroviral vector and the pVSVg plasmid were transfected into eukaryotic cells 293T by calcium phosphate transfection method. And then, the pseudovirus were produced. The pseudovirus infected eukaryotic cells SP2/0 and E1E2 protein was expressed. E1E2 protein was detected by puromycin-resistant and FACS analysis. BALB/c mice were injected in abdomen with expressing E1E2 protein SP2/0 cells. Anti-HCV E1E2 antibody was screened by FACS. Moreover, the antibody was also analyzed by Western blot using E2 protein antigen which was expressed in E. coli. The results showed that HCV E1E2 protein was expressed in SP2/0 cells’ envelope successfully. FACS could detect specific anti-E1E2 antibody in SP2/0 cells immune mouse serum. Western blot analysis showed that SP2/0 cells immune mouse serum could react specially to E2 protein that was expressed in E.coli.