Construction of Recombinant Plasmid and Expression of Surface Glycopro- tein gp120 of HIV-1

Objective To construct an eukaryolic expression plasmid and express surface glycoprotein gp120 gene of HIV-1 in Hela cell. Methods An eukaryotic expression plasmid pVAXGP was constructed by inserting gp120 gene into DNA vaccine vector plasmid pVAX1, and transfected to Hela cell in vitro by lipo-some-mediated method. The expressed product was detected by indirect immunofluorescence assay(IFA), Western blot and Dot-ELISA. Results Indirect IFA showed green fluorescence on the surface of transfected cells.Both Western blot and Dot-ELISA proved that the lysate of transfected cells contained gp120 protein, Conclusion An eukaryotic expression plasmid for gp120 gene of HIV-1 was successfully constructed, and the expressed product showed good reactogenicity and specificity.