Abstract: The beta-adrenergic receptors (β-AR) are a class of metabotropic G protein-coupled receptors that responsible for the muscle growth, fat deposition or lactation characteristics of domestic animals. The aim of this article was to scan nucleotide mutations in partial coding region of β2-AR gene on cattle. In this study, the polymorphisms of β2-AR gene on cattle were detected by methods of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and DNA sequencing in two cattle breeds in China, Xinjiang Brown cattle and Chinese Holstein cows. The results revealed that there were two genotypes and homozygote variant was not found in the two breeds. The genotypic frequencies at the Sma I locus were significantly different between the two breeds. The wild-type allele was ascendancy in the two breeds but its allele frequency was distinctly lower in the Chinese Holstein cows than that in the Xinjiang Brown cattle. DNA alignment results showed that three single nucleotide polymorphisms (SNPs), C11A, G53A and C129T were found in 5-coding region of β2-AR gene. The C11A mutation resulted in disappearance of Sma I cleavage site. In addition, the C11A and G53A mutations caused two amino acid residues replacement at the 4th and 18th of β2-AR. However, the C129T mutation was a synonymous one. In conclusion, the heterozygous frequency in Chinese Holstein cows was obviously higher than that in the Xinjiang Brown cattle. The Sma I locus would be a potential genetic marker for lactation performance of cattle.