FIG. 4.
PRP4 is not incorporated into HIV-2 virus particles. (A) Schematic diagram of HIV-2 Gag and PRP4 expression constructs. CMV, cytomegalovirus immediate-early inhancer; T7, promoter/priming site; MCS, multiple cloning site; V5, V5 epitope; His6, polyhistidine tag; RS, arginine-serine-rich domain; KS, kinase domain. (B and C) 293T cells were transfected with PRP4 plasmid (pCDNA3.1V5PRP4FL) or control plasmid (pCDNA3.1V5HisA) and an env-deleted HIV-2 provirus as indicated. Cell and virus particle samples were prepared as described in Materials and Methods and subjected to analysis by Western blotting using monoclonal antibodies to either p26 (HIV-2 capsid) (B) or V5-tagged PRP4 (C). The positions of marker proteins are indicated in kilodaltons.