Molecular cloning and biological characterization of full-length HIV-1 subtype C from Botswana

Human immunodeficiency virus type 1 (HIV-I) subtype C is now responsible fo r more than half of all HIV-1 infections in the global epidemic and for the high levels of HIV-1 prevalence in southern Africa. To facilitate studies of the biological nature and the underlying molecular determinants of this virus, we constructed eight full-length proviral clones from two asymptomat ic and three AIDS patients infected with HIV-1 subtype C from Botswana. Ana lysis of viral lysates showed that Gag, Pol, and Env structural proteins we re present in the virions. In four clones, the analysis suggested inefficie nt envelope glycoprotein processing. Nucleotide sequence analysis of the ei ght clones did not reveal frameshifts, deletions, premature truncations, or translational stop codons in any structural, regulatory, or accessory gene s. None of the subtype C clones were replication competent in donor periphe ral blood mononuclear cells (PBMCs), macrophages, Jurkat(tat) cells, or U87 .CD4.CCR5 cells. However, infection by two clones could be rescued by compl ementation with a functional subtype C envelope clone, resulting in a produ ctive infection of PBMCs, macrophages, and U87.CD4.CCR5 cells. (C) 2000 Aca demic Press.