title: Construction, expression, purification and antigenicity of recombinant Campylobacter jejuni flagellar proteins.
abstract: Campylobacter jejuni, a flagellated, spiral-rod Gram-negative bacterium, is the leading etiologic agent of human acute bacterial gastroenteritis worldwide. The source of this microorganism for human infection has been implicated as consumption and handling of poultry meat where this microorganism is a commensal in the gut. Because the genomes of many C. jejuni isolates have been sequenced, our ultimate goal is to develop protein arrays for exploring this microorganism and host interactions. In this communication, we report cloning, expression and purification of C. jejuni flagellar proteins in a bacterial expression system. Twelve recombinant proteins were purified, which were confirmed by SDS-PAGE analysis and a His tag detection kit. The FlgE1, FlgG, FlgK, FliE, FlgH/FliH and FlaA recombinant proteins were further confirmed by LC-ESI-MS/MS. The purified recombinant proteins were tested whether they were immunogenic using antibodies from several sources. BacTrace anti-Campylobacter species antibody reacted to the FlaA recombinant protein, but not others. Rabbit anti-MOMP1 peptide antibody reacted strongly to FliE and weakly to FlaA, but not others. Rabbit anti-MOMP2 peptide antibody reacted strongly to the FlaA, FliG, FliE, FlhF, FlgG, FlgE1 and FliD recombinant proteins, less to FlgK and FlgH/FliH, and did not react to the FliY, FliS and FliH recombinant proteins. These antibody studies suggest that these recombinant flagellar proteins have potential for novel targets for vaccine development. It is also anticipated that these recombinant proteins provide us a very useful tool for investigating host immune response to C. jejuni.
authors: United States 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. hungyueh.yeh@ars.usda.gov
authors: Hung-Yueh
authors: HY
authors: Yeh
authors: Kelli L
authors: KL
authors: Hiett
authors: John E
authors: JE
authors: Line
authors: Brian B
authors: BB
authors: Oakley
authors: Bruce S
authors: BS
authors: Seal
mesh_terms: D000818:Animals; D002169:Campylobacter Infections / Q000382:microbiology; D016123:Campylobacter jejuni / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D002645:Chickens; D005243:Feces / Q000382:microbiology*; D005407:Flagella / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D005408:Flagellin / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D053719:Tandem Mass Spectrometry
chemical_list: D011994:Recombinant Proteins; D005408:Flagellin