Abstract: To further study the regulatory mechanisms of Cholesterol 7α-hydroxylase (CYP7A1) in bile acid synthesis and cholesterol metabolism, the complete cDNA of CYP7A1 gene(2 279 bp) was cloned by RT-PCR and RACE from goose (Anser anser) liver. The CDS of CYP7A1 gene was sub-cloned into pET-28a to construct pET-28a-cyp7α1 plasmid. The recombinant plasmid pET-28a-cyp7α1 was transformed into E.coli BL21 (DE3) and the recombinant protein expression was induced by IPTG. The recombinant protein His-CYP7A1 was purified with His-Bind Purification Kit. The recombinant protein was immunized into the Bal b/c mice to obtain the polyclonal antibodies. Antigenic of His-CYP7A1 and the specificity of mouse anti-goose CYP7A1 polyclonal antibodies were detected by Western blotting. The amino acid sequence of goose CYP7A1 protein has high homology with that of chicken(93%), rat(66%), human(67%), monodelphis domestica(68%) and mouse(66%). The SDSPAGE analysis result of His-CYP7A1 showed that the recombinant protein molecular size was about 59 ku, consistent with the predicttion. Highpurity His-CYP7A1 protein was obtained through His-tag affinity purification.The mouse anti-goose CYP7A1 polyclonal antibodies were obtained with high sensitivity (1∶10⁴). The mouse anti-his and anti-goose CYP7A1 antibodies could recognize antigens of His-CYP7A1 by Western blotting with mouse anti-goose CYP7A1 polyclonal antibodies with better specificity. The current results contribute to further understanding of gene structure and function of CYP7A1 and provide an important molecular tool for the functional study of CYP7A1.