Genetic comparison of SIVmac to the human retroviruses generally associated with AIDS revealed a closer relationship to HIV-2 than to HIV-1. A common feature differentiating SIV and HIV-2 from HIV-1 is the size of the transmembrane portion of the envelope, which is smaller (gp32) in SIVmac and HIV-2 than in HIV-1 (gp41). The presence of this truncated form of the transmembrane glycoprotein in SIVmac and HIV-2 virions is apparently related to the presence of a translation termination codon in the env gene of all SIV proviruses analyzed as well as in one HIV-2 provirus. Since the carboxy terminus of the envelope transmembrane protein has been implicated in the cytopathic effect of HIV-1 in vitro, we decided to investigate whether putative expression of the open reading frame located after the termination codon correlates with the pathogenicity of SIVmac in vivo. We generated two synthetic peptides from the inferred amino acid sequence of SIVmac and tested their reactivity by Western blot against the sera of naturally and experimentally infected monkeys as well as against sera of HIV-2-infected individuals. Our results indicate that the protein synthesized from this open reading frame is expressed in vivo, since an immunoresponse can be detected against the synthetic peptides in two of three experimentally SIVmac-infected animals. However, no correlation can be found between its expression and disease progression at this time. Furthermore, a rabbit immune serum raised against the synthetic peptide failed to identify any specific protein in SIVmac-infected cells.