To　prepare　HIV-1　Vif　and　hAPOBEC3G　and　to　produce　their　antibodies,　the　full　length　gene　fragment　of　HIV-1　vif　was　amplified　by　PCR　from　a　plasmid　of　HIV-1　NL4.3　cDNA,　and　the　APOBEC3G　gene　was　obtained　by　RT-PCR　from　the　total　RNA　of　H9　cells.　The　resulting　DNA　construct　was　cloned　into　a　prokaryotic　expression　vector　(pET-32a).　Recombinant　pET-vif　and　pET-APOBEC3G　were　expressed　respectively　in　Eserichia　coli　BL21　(DE3)　as　an　insoluble　protein.　The　vector　also　contained　a　six-histidine　tag　at　the　C-terminus　for　convenient　purification　and　detection.　To　express　and　purify　the　HIV-1　Vif　and　hAPOBEC3G　in　E.coli　cells,　the　accuracy　of　inserted　gene　and　specificity　of　proteins　were　detected　by　the　two　enzyme　digestion　method,　SDS-PAGE,　and　Western　blotting.　Rabbits　were　then　immunized　by　Vif　or　APOBEC3G　protein　and　serum　samples　were　tested　by　indirect　ELISA　to　determine　the　level　of　antibodies.　Immunoenzyme　and　immunofluorescence　assays　were　performed　to　identify　the　specificity　of　polyclonal　antibodies.　The　titer　of　the　anti-Vif　antibodies　was　1:204800,　and　that　of　the　anti-APOBEC3G　antibodies　was　1:102400.　Thus　the　antibodies　could　detect　the　antigen　expression　in　the　cells,　demonstrating　that　fusion　proteins　with　high　purity　and　their　corresponding　polyclonal　antibodies　with　high　titer　and　specificity　were　achieved.　©　Wuhan　Institute　of　Virology,　CAS　and　Springer-Verlag　GmbH　2008.