The Rhesus (Rh) gene superfamily in humans and mice contains four independent genes, RH, RHAG, RHBG, and RHCG/GK. Heretofore, only the RHBG cDNA has been cloned in pig. We have isolated the porcine RH cDNA; its complete open reading frame of 1269 nucleotides encoded 423 amino acids. Porcine RH protein shared 67.6% amino acid identity with bovine RH, 61.0% with human RhCE and 60.8% with human RhD. The RT-PCR revealed RH transcripts in the spleen and bone marrow, but not in the heart, kidney, or lung. In RH intron 4, a deletion of 17 nucleotides distinguished the shorter allele (allele 1) from the longer. As determined in 115 unrelated pigs from five breeds - Landrace (L, n = 23), Large White (LW, n = 28), Duroc (D, n = 24), Hampshire (H, n = 20) and Piétrain (n = 20) - allele 1 frequencies were 1.0 (L, H), 0.77 (LW), 0.70 (P) and 0.25 (D). Somatic cell hybrid mapping localized the porcine RH and RHBG genes to pig chromosomes 6q22-q23 and 4q21-q22, respectively. Genetic mapping suggested RH-(FUT1, S, GPI, EAH, A1BG)-PGD as the most probable locus order. Sequence homology, mapping data, and haematopoietic tissue expression suggest that this cDNA may indeed encode the porcine RH homologue.