PREPARATION METHOD OF PROTECTIVE ANTIGEN USING TRANSFORMED BACILLUS WITH RECOMBINANT VECTOR INTRODUCED PROTECTIVE ANTIGEN

摘要

PURPOSE: Recombinant protective antigen of anthrax is over expressed using Bacillus brevis 47-5Q and pNU212 vector system and then purified protective antigen is used for vaccine. CONSTITUTION: PCR is performed to amplify protective antigen of anthrax and cloned into T -vector. Cloned insert is isolated after restriction enzyme digestion and ligated with pNU212 vector. The resulting vector pNU212-PA is transformed into Bacillus brevis 47-5Q and the supernatant of culture is analyzed using western blotting. Purification of protective antigen secreted into medium is achieved using FPLC. Purified protective antigen is confirmed by checking the molecular weight of protein ran in the SDS-PAGE, western blotting, and immunoblotting. N-terminal amino acid sequence of expressed gene is compared with the protective antigen produced from standard strain Bacillus anthracis ATCC14185.