Abstract: Further study on FecB gene,the complete FecB gene was amplified with PCR method using a pair of specific primers designed according to the relevant nucleotide sequence from GenBank.And FecB gene was cloned into vector pET30a(+) for expressed in E.coli BL21(DE3).The expression of 6×His and FecB fusion protein was induced by IPTG,then purified by Ni-NTA chromatographic method.Polyclonal antibodies were prepared by immunized mice with the purified recombinant protein.Expression of the target protein was detected by SDS-PAGE,the specificity and titer of the antibody in anti-sera was determined by Western Blot and ELISA respectively.The recombinant FecB can be expressed by IPTG induction and purified by Ni-NTA resin.The results of ELISA and Western Blot proved that polyclonal antiserum prepared with purified recombinant FecB as antigen has high titration(>1: 32 000)and specificity.A method for prokaryotic expression and purification of sheep FecB was established and the anti-FecB polyclonal antibody with high titration and specificity were obtained.These results would provide reliable tools for the future study on FecB function.

Key words: FecB gene, Prokaryotic expression, Protein purification, Polyclonal antibody

CLC Number: 

• Q78