2003•Biotechnology(Faisalabad)Requires access
Construction of Recombinant Plasmid and Expression of Fusion of HIV-1gag/IFNα-2b
Jiqun Wang
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Abstract
Objective:To construction an eukaryotic expression plasmid and express fusion of HIV-1gag/IFNα-2b in BHK21cell.Methods:An eukaryotic expression plasmid pSFJ38gag/IFNα-2b was constructed by inserting IFNα-2b gene into site (nt531) of gag gene and inserting HIV-1gag/IFNα-2b gene into vaccine vector plasmid pSFJ38,and transfected to BHK21 cell in vitro by liposome-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 cell.Both Western blot and Dot-ELISA proved that the lysate of transfected cells contained gag/IFNα-2b protein.Conclusion:An eukaryotic expression plasmid for HIV-1gag/IFNα-2b was successfully constructed,and the expressed product showed good reactinogenicity and immunogenictiy.