The MC1R gene is related to coat color in cattle and to lactoprotein content in milk. Polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing were used to analyze the MC1R of China Holstein Black-white Cattle, China Holstein Red-white Cattle, Luxi Yellow Cattle and Bohai Black Cattle. Three alleles (ED, E+ and e) were identified. Most China Holstein Black-white Cattle had ED and E+ alleles (ED = 0.12, E+ = 0.80). The same was true for Bohai Black Cattle (ED = 0.52, E+ = 0.47). On the other hand, most China Holstein Red-white Cattle and Luxi Yellow Cattle had the e allele (e/e = 0.95). E+/e genotype was present in China Holstein Red-white Cattle and Luxi Yellow Cattle. We reasoned that ED and E+ lad to the synthesis of the black pigment. Furthermore, we found an important SNP (Single Nucleotide Polymorphism) at 725 position of the MC1R coding region.