Prokaryotic Expression and Polyclonal Antibody Preparation of BLCAP

Objective:To express BLCAP fusion protein in E.coli BL21(DE3) and prepare polyclonal antibody. Methods:The prokaryotic expression plasmid pET32a(+)-BLCAP was constructed and the positive recombinant plasmid was transformed into BL21(DE3) and induced by IPTG. After the soluble protein was purified by affinity chromatography and identified by SDS-PAGE,the rabbits were immunized with the fusion protein and the antiserum was obtained. The specificity of the rabbit’s antiserum was measured by Western bloting. Results:Nearly 28 ku fusion protein was successfully expressed after being induced by IPTG. The polyclony antibody was also successfully prepared and its specificity was proved by Western blotting. Conclusion:The BLCAP fusion protein has a good antigenicity and the BLCAP polyclonal antibody has a high specificity,which set the foundation for the study of the characters and function of BLCAP protein.