The bovine immunodeficiency-like virus (BIV) is morphologically, serologically, and genetically related to the lentivirus subfamily of retroviruses which includes human and simian immunodeficiency viruses and other lentiviruses causally associated with debilitating diseases of domestic animals. There are many parallels in the biology and pathologic characteristics of BIV infections with those of HIV that make its development as a model of HIV-like infection and disease potentially attractive. In order to obtain a better understanding of the molecular basis of BIV-induced disease, two biologically active proviruses of BIV were molecularly cloned and sequenced. The BIV genome is 9.0 kilobases in the form of the proviral DNA. It contains the obligate retroviral structural genes, gag, pol, and env. In addition, in the BIV central region, between and overlapping pol and env, there are five potential coding regions for non-structural/regulatory genes; three are analogous to vif, tat, and rev in HIV and two, called W and Y, are unique to BIV. There is no coding region analogous to nef in BIV. Sequence comparisons of two functional proviruses obtained from the DNA of cells carrying an infection from a single virus isolation indicate that the genome of BIV is highly variable within a single biological isolate. Moreover, the greatest number of substitutions occur in the env gene. The results suggest the presence of multiple genotypes which may be of significance in defining the disease potential of a BIV isolate. These clones will be useful in dissecting the replicative cycle and mechanisms of pathogenesis of BIV in various animal models.