Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus

【摘要】 The Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals.Bacteria flagellins play an important role during infection and induction of the host immune response.Thus,flagellin proteins are an ideal target for vaccines.We amplified the complete flagellin subunit gene(flaA) from V.parahaemolyticus ATCC 17802.We then cloned and expressed the gene into Escherichia coli BL21(DE3) cells.The gene coded for a protein that was 62.78 kDa.We purified and characterized the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting,respectively.Our results provide a basis for further studies into the utility of the FlaA protein as a vaccine candidate against infection by Vibrio parahaemolyticus.In addition,the purified FlaA protein can be used for further functional and structural studies.更多还原

【Abstract】 The Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals.Bacteria flagellins play an important role during infection and induction of the host immune response.Thus,flagellin proteins are an ideal target for vaccines.We amplified the complete flagellin subunit gene(flaA) from V.parahaemolyticus ATCC 17802.We then cloned and expressed the gene into Escherichia coli BL21(DE3) cells.The gene coded for a protein that was 62.78 kDa.We purified and characterized the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting,respectively.Our results provide a basis for further studies into the utility of the FlaA protein as a vaccine candidate against infection by Vibrio parahaemolyticus.In addition,the purified FlaA protein can be used for further functional and structural studies.更多还原