Construction and expression of recombinant adenovirus carrying HIV-1 vpr gene

Objective To construct the recombinant adenoviral vector expressing HIV-1 vpr gene and to induce viral protein R(Vpr)highly expressed in C8166 cells.Methods The HIV-1 vpr gene was cloned to the shuttle plasmid pAdTrack-CMV.The linearized resultant plasmid and adenoviral backbone plasmid pAdEasy-1 were co-transfected into E.coli BJ5183.Recombinant adenoviral plasmids were screened and identified,the linearized recombinant plasmid was transfected into HEK293A for adenovirus package through lipofectamine 2000.Viral productions were monitored by GFP expression.Results The results shown that the recombinant adenovirus carrying HIV-1 vpr gene was constructed successfully.High expression of HIV-1 vpr gene in C8166 was detected by Western blotting and the high infection rate was detected by FACS.Conclusion Recombinant adenovirus Ad-vpr is successfully constructed and the Vpr protein is specially expressed in C8166 cells.