Abstract: The high-fidelity PCR method had been used to amplificate 1.8 and 1.1 kb 5′and 3′ regulatory sequences of bovine casein gene in mammary grand. Then it was cloned into TA vector. Veritficated by the restriction enzyme digestion and PCR method，after sequence on NCBI with blast，the results indicated that the tragments have the homology of 97.0% and 99.0% respectively with the correspondingly region of boving beta-casein. It is discribed that the restructuring carrier had succeeded in cloning the area of control casein gene 5′and 3′, the recombinant DNA technology had been used to subclone into the modified eukaryotic expression vector pcDNA3 (removal of CMV promoter)，verificated by the restriction enzyme digestion and sequence analysis，the mammary grand-specific expression vector of cattle had been constructed.

Key words: beta-casein gene; cloning; cDNA; mammary grand-specific expression