Expression of CSFV E2 Envelope Protein Gene in Eukaryotic Cells and Preparation of Monoclonal Antibodies against CSFV E2 Envelope Protein
Abstract: The recombinant retroviral vector pBABE-puro-E2 was constructed by inserting full-length cDNA of CSFV Shimen strain E2 gene into pBABE-puro. Both the recombinant retroviral vector and pVSVg plasmid were transfected into eukaryotic cells 293GP by calcium phosphate transfection method, and the pseudovirus were produced. The pseudovirus-infected eukaryotic cells PK-15 and expression of E2 protein were determined by puromycin-resistant and FACS analysis. Balb/c mice were intraperitoneal injected with PK-15 cells expressing the CSFV E2 protein. Anti-CSFV E2 antibody was screened by ELISA. The results showed that CSFV E2 protein was expressed in PK-15 cells’ envelope protein successfully. ELISA could detect specific anti-E2 antibody in mouse serum immuned by PK-15 cells.