FIG. 4.
Analysis of wild-type and ts mutant viral proteins produced in transfected HeLa cells. HeLa cells were transfected with the indicated plasmids (wt, HIV-1LAI proviral DNA; mt, mt4 proviral DNA; PR−, PR-defective proviral DNA; PR−/mt, mt4 proviral DNA with a mutated PR; mock, pUC19). (A) Forty-eight hours posttransfection, viral particles were harvested from the cells incubated at either 34.5 or 39.5°C. Virion-associated proteins were analyzed by immunoblotting using the anti-HIV-1 RT polyclonal antibody described in Materials and Methods. (B) Forty-eight hours posttransfection, the HeLa cells were metabolically labeled with [35S]Met at 39.5°C for 12 h. Cell-associated or virion-associated proteins were immunoprecipitated with an AIDS patient serum and resolved on SDS–10% polyacrylamide-AcrylAide gels. The markers used are described in the legend to Fig. 2.