Cloning and expressing of HIV-1 Vif gene in prokaryotic cells, and evaluation of its bioactivity.

In order to clone HIV-1 Vif gene,construct prokaryotic expression plasmid and detect its expression and biological activity,HIV-1 Vif gene was obtained by PCR and cloned into the vector pET28a(+)to construct prokaryotic expression plasmid pET28a(+)/Vif.The positive plasmid was identified by double enzyme digestion and sequencing.Positive plasmid was transformed to E.coli BL21(DE3)and the expressed product was identified by SDS-PAGE electrophoresis and western blot analysis.Vif protein was expressed and purified,and its biological activity was assayed.Results indicated that the plasmid pET28a(+)/Vif was correctly constructed.Through transformation of pET28a(+)/Vif into E.coli BL21(DE3),the Vif protein was expressed and purified.The renatured protein showed that Vif could bind into SH3(HCK)domain in the pull-down assay.It's concluded that the successful cloning,expression and purification of Vif protein provide the basis for further studies on the functions of the protein.