Isolation and characterization of a full-length molecular DNA clone of Ghanaian HIV type 1 intersubtype A/G recombinant CRF02_AG, which is replication competent in a restricted host range

We have isolated a replication-competent, full-length molecular clone of HI V-1 CRF02_AG, designated p97GH-AG1, by reconstituting two separately amplif ied genomic regions of an HIV-1 provirus of a 1997 Ghanaian isolate. The ph ylogenetic and recombination breakpoint analyses revealed that 97GH-AG1 had an A/G recombinant structure similar to that of prototype Nigerian isolate IbNG. The 17-nucleotide insertion downstream of the primer-binding site ap peared to be a common sequence signature specific to most CRF02_AG strains, including 97GH-AG1. 97GH-AG1 showed an R5 phenotype and exerted productive infection in both HOS and NP2 cell infectivity assays, whereas it failed t o show a detectable level of progeny production in peripheral blood mononuc lear cells (PBMCs), The data may suggest the presence of unknown determinan t(s) that dictate efficient replication in PBMCs, but that are not required for replication in immortalized cell lines.