Expression and Cloning of Partial flaA Gene of Legionella Pneumophila in Prokaryotic Cell

Objective To clone and express the recombinant partial flagellum subunit gene of Legionella pneumophila,and to purify the recombinant protein.Methods The partial flagellum subunit gene of Legionella pneumophila was amplified from the total DNA of Legionella pneumophlia serogroup 1 by polymerse chain reaction and then was cloned into prokaryote expression vector pET32a(+).After the recombinant plasmid was identified by PCR,restriction enzyme analysis and sequencing analysis,the recombinant plasmid pET-flaA was constructed and transferred into E.coli strain BL21.The expression of fusion protein was induced with isopropy-β-D-thiogalactoside(IPTG) and examined with SDS-PAGE,and was purified by Affinity chromatography.Results The partial flaA gene of 606 bp in length was amplified.The recombinant plasmid pET-flaA was expressed and purified 42 kDa fusion protein.Conclusion The recombinant plasmid containing Lgeionella pneumophila the partial flaA gene constructed in this study is highly efficient expression in prokaryotic cell.