We have studied the variability of four structurally complete dog leukocyte antigen (DLA) class I genes, termed DLA-12, -88, -79 and -64, in a population of mixed breed, unrelated dogs. The human HLA and canine DLA loci share a high degree of similarity in terms of gene structure. This analysis focused on the first three exons of each of four complete canine genes. Exons two and three are the major source of polymorphism in the corresponding human genes. In this analysis, DLA-88 was found to be significantly more polymorphic than the other three genes, with 44 distinct alleles observed among 63 mixed breed, unrelated dogs. The remaining genes had between one and four alleles when examined in 25 dogs. This work was carried out as part of an effort to develop an MHC typing system for the dog, which is critical to the further development of preclinical studies of hematopoietic stem cell and solid organ transplantation in the canine model.