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A previous genetic analysis of HIV-1 strains circulating in Lisbon, Portugal, demonstrated a high prevalence of non-B subtypes, with a predominance of subtype G, CRF02_AG and CRF14_BG [3, 4]. In order to assess natural resistance mutations to PI in HIV-1 subtype G, we analysed protease and cleavage site amino acid substitutions in 52 antiretroviral therapy-naive individuals, 27% of whom were of African origin, half of them having reported injecting drug use. Previous phylogenetic characterization of env sequences and gag subtyping by heteroduplex mobility assay [3, 4] had assigned 38 of the infecting viruses to subtype G, eight to CRF14_BG, and six to CRF02_AG. A 600 base pair fragment of proviral DNA, covering the 39 end of gag and the protease coding sequence, was amplified from peripheral blood mononuclear cell lysates by nested polymerase chain reaction, with primer sets P1F, 59 …