The complete coding sequences of three of the Black-boned sheep (Ovis aries) genes - RhoB, RhoF and RhoH were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) according to the conserved sequence information of the mouse or other mammals and known highly homologous sheep ESTs. The Black-boned sheep RhoB gene encodes a protein of 196 amino acids which contains the conserved putative RhoA_like domain and has high homology with the RhoB proteins of six species-cattle (100%), goat (100%), human (100%), pig (100%), mouse (100%) and chicken (97%). The Black-boned sheep RhoF gene encodes a protein of 215 amino acids that contains the conserved putative Rho4_like domain and has high homology with the RhoF proteins of five species-bovine (100%), human (94%), pig (93%), mouse (92%) and chicken (58%). The Black-boned sheep RhoH gene encodes a protein of 191 amino acids that contains the conserved putative Rho domain and has high homology with the RhoH proteins of four species-bovine (99%), human (96%), pig (96%) and mouse (96%). The phylogenetic tree analysis demonstrated that the Black-boned sheep RhoB, RhoF and RhoH proteins share a common ancestor. The tissue expression analysis indicated that the three genes were expressed in a range of tissues including small intestine, large intestine, liver, muscle, fat, lung and spleen. Our experiment is the first to provide the primary foundation for further insight into these three sheep genes.