Abstract: Sex of breeded animal is a very important character for stockbreeding, while for mammal, the sex determination is started by gene Sry. So, to obtain SRY protein and study it’s interaction with other correlative genes plays an important role in making up efficient and economic technique for sex control of breeded animal.In this experiment, we used PCR、plasmid recombination、restriction digest and sequencing to construct pET-21b-Sry for sex determining gene of mice – Sry and detect it’s validity. While, we adopt protein expression induced by IPTG、 SDS-PAGE and western-blotting to achieve and detect SRY protein from prokaryotic expression. All the results of PCR、plasmid recombination、restriction digest and sequencing displayed Sry fragement from mice and prokaryotic expression vector pET-21b-Sry were abteined succesfully. Then, the validity of Sry-His protein in BL21(DE3) was proved with western blotting using anti-Sry and anti-His. In conclusion, Prokaryotic Expression for Sry-His was carried out in this experiment.