Prokaryotic Expression of FecB Gene and Generation of Its Polyclonal Antibody

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 Gen Bank. And FecB gene was cloned into vector p ET30a( +) for expressed in E. coli BL21( DE3). The expression of 6 × His and Fec B 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 Fec B 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 Fec B as antigen has high titration( 1 ∶ 32 000) and specificity. A method for prokaryotic expression and purification of sheep Fec B was established and the anti-Fec B polyclonal antibody with high titration and specificity were obtained. These results would provide reliable tools for the future study on Fec B function.