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We previously demonstrated that serine protease inhibitor BmSPI could strongly inhibit subtilisin A.In order to characterrize the localization of BmSPI,the recombinant protein was expressed in Escherichia coli BL21 (DE3) Star.The recombinant BmSPI was injected into the hypodermis of New Zealand white rabbit for producing polyclonal antibody,after affinity purification combined with reversed-phase chromatography.