To examine the factors that control the extent of incorporation of Vpr into the virus particles, we utilized an epitope-tagging approach with Flag (FL) as the epitope for quantitation. We generated expression plasmids containing Vpr-FL and Vpr E21,24P-FL and also HIV-1 proviral DNA containing Vpr-FL (NL-Vpr-FL). Immunoblot analysis using Flag antibodies revealed that virus particles derived from co-transfection of NL-Vpr-FL and Vpr-FL showed an enhanced level of Vpr-FL in comparison to NL-Vpr-FL derived virus. These results suggest that the amount of incorporation of Vpr into the virus particles is flexible and may be modulated by its expression level in cells.
