Recombinant expression，purification and adhesion function identify of Bacillus anthracis BslA(260－652) protein

Abstract:

Abstract:［Objective］To obtain the recombinant BslA(260－652) protein of Bacillus anthracis and prepare its antibody for the adhesion activity studies．［Methods］The fragment coding BslA(260－652) was cloned into pET28a(+) plasmid and induced to express recombinant protein in E． coli Rosetta (DE3) by Isopropyl β-D-1-thiogalactopyranoside (IPTG)．The expressed recombinant soluble protein was purified by a column packed with Ni Resin． Purified protein was used as the antigen to immunize BABL/c mice for three times to raise polyclonal antibody． The adhesion activity of BlsA(260－652) was detected by immunofluorescence experiments and bacterial adherence assays．［Results］The purity of the purified soluble BslA(260－652) was about 87.4%．ELISA assay titer of antiserum from vaccinated mice reached 1:20000. Western blot showed the antiserum could specifically recognize endogenous BslA protein． The purified BslA(260－652) displayed a typical adhesion-like function． Either the anti-BslA serum or the BslA(260－652) protein could inhibit A16R's Hela adherence．［Conclusion］The recombinant BslA(260－652) protein was successfully obtained，which would lay the foundation for further research of the anthrax vaccine and the role of this S-layer protein in the pathogenesis of anthrax.