Abstract: This study was aimed to express and purify the flagellins, and contribute to research and gain high efficient adjuvants. The fljB, fljB' and fliC genes were amplified from Salmonella Typhimurium genomic DNA by PCR, and the amplified PCR products were cloned into pMD19-T Simple Vector to construct cloned plasmid pMD19-fljB, pMD19-fljB'and pMD19-fljC. The cloned plasmids were digested by double enzymes, and purified genes were subcloned into pET-30a vector to construct prokaryotic expression vector pET30a-fljB, pET30a-fljB'fliC and pET30a-fliC. Transformed colonies were screened by PCR, restriction enzyme analysis and sequencing. Then the positive expression vectors were transformed into E. coli BL21(DE3) strain. After induced by IPTG, the expressed proteins were extracted from E. coli BL21(DE3) strain using Ultrusonic Cell Disrupter System. The fljB, fljB' fliC and fliC were purified by Ni-NTA Columns. The recombinant proteins were further confirmed by Western blotting analysis using anti-flagellin serum from Guinea pig. Through optimizing the expression conditions, flagellins were expressed in dissoluble form. The purified flagellins would lay the foundation for researching their adjuvant properties later.

Key words: flagellin; prokaryotic expression; purification