FIG. 1.
Immunoprecipitation analysis of HIV-2ALI envelope glycoproteins expressed by recombinant vaccinia viruses. (A) Schematic representation of the pMJALI plasmid used to produce rVV/ALI. The env gene from HIV-2ALI was cloned into the unique SalI site of the pMJ601 plasmid, which is located adjacent to the synthetic late promoter within the X region (16). (B) HIV-2ALI and HIV-2ROD envelope glycoproteins expressed in HeLa cells by recombinant vaccinia viruses rVV/ALI and rVV/ROD (positive control), respectively. (C) Schematic representation of gp125t protein expressed by rVV/ALIM2, with the sizes and approximate locations of the conserved and variable domains indicated. (D) Pulse-chase analysis of gp125t expression. HeLa cells infected with rVV/ALIM2 were metabolically labeled for 30 min with [35S]methionine and chased for 1, 3, and 18 h. In panels B and D, viral glycoproteins were immunoprecipitated from cell lysates (E) and supernatant (S) with antiserum from an HIV-2-infected individual and analyzed by SDS-PAGE and fluorography. Mock, uninfected cells; vWR, cells infected with the WR strain of vaccinia virus (negative control). Standard molecular mass markers are indicated in kilodaltons.