Characterization and antigenicity of recombinant Campylobacter jejuni flagellar capping protein FliD.
Campylobacter jejuni, a flagellated, spiral-rod, Gram-negative bacterium, is the leading pathogen of human acute bacterial gastroenteritis worldwide, and chickens are regarded as a major reservoir of this micro-organism. Bacterial flagella, composed of more than 35 proteins, play important roles in colonization and adhesion to the mucosal surface of chicken caeca. In this study, the flagellar capping protein, FliD, encoded by the fliD gene, from the Campylobacter jenuni D1-39 isolate was expressed and characterized, and its antigenicity determined. The fliD gene comprised 1929 nt, potentially encoding a 642 aa peptide with a calculated molecular mass of 69.6 kDa. This gene was PCR amplified and overexpressed in Escherichia coli. The recombinant FliD protein was purified by cobalt-chelating affinity chromatography and confirmed by nucleotide sequencing of the expression plasmid, SDS-PAGE analysis, His tag detection and matrix-assisted laser desorption/ionization time of flight mass spectrometry. The immunoblot data showed that the purified recombinant FliD protein reacted strongly to sera from broiler chickens older than 4 weeks, indicating that anti-FliD antibody may be prevalent in the poultry population. These results provide a rationale for further evaluation of the FliD protein as a vaccine candidate for broiler chickens to improve food safety for poultry.
US Department of Agriculture, Agricultural Research Service, Richard B. Russell Agricultural Research Center, Poultry Microbiological Safety Research Unit, 950 College Station Road, Athens, GA 30605-2720, USA.
Hung-Yueh
HY
Yeh
US Department of Agriculture, Agricultural Research Service, Richard B. Russell Agricultural Research Center, Poultry Microbiological Safety Research Unit, 950 College Station Road, Athens, GA 30605-2720, USA.
Kelli L
KL
Hiett
US Department of Agriculture, Agricultural Research Service, Richard B. Russell Agricultural Research Center, Poultry Microbiological Safety Research Unit, 950 College Station Road, Athens, GA 30605-2720, USA.
John E
JE
Line
US Department of Agriculture, Agricultural Research Service, Richard B. Russell Agricultural Research Center, Poultry Microbiological Safety Research Unit, 950 College Station Road, Athens, GA 30605-2720, USA.
Bruce S
BS
Seal
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000235:genetics* / Q000276:immunology*; D002645:Chickens; D002846:Chromatography, Affinity; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D015151:Immunoblotting; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; C076733:FlaD protein, Bacteria