Cloning of an Outer Membrane Protein Gene from Campylobacter jejuni |
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Authors: | Richard J Meinersmann Kelli L Hiett Alacia Tarplay |
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Institution: | (1) USDA, Agricultural Research Service, Poultry Microbiological Research Unit, P.O. Box 5677, Athens, GA 30604-5677, USA , US |
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Abstract: | An antigen in the outer membrane protein (OMP) fraction of
Campylobacter jejuni was identified and characterized. Western blot
analysis demonstrated antigenic differences in this protein between two
congenic C. jejuni strains. Strain A74/C, which colonizes chickens,
expressed the antigen at 34 kDa, while strain A74/O, which poorly colonizes
chickens, expressed the antigen at 32 and 34 kDa. A genomic library was
constructed in λgt11 with DNA from A74/O and screened with antibody raised
against C. jejuni OMPs. A clone that possessed a 1.3-kb insert and
expressed an immunoreactive protein fused to β-galactosidase was isolated
and purified. DNA sequence analysis revealed the insert contained one open
reading frame 864 bases long. The deduced amino acid sequence demonstrated
56.3% similarity with Bacillus steorothermophilus glnH, a
glutamine-binding protein, and 54.0% similarity with C. jejuni
PEB1, a putative colonization adhesin. Southern hybridization, Northern
hybridization, and DNA sequence analyses of the congenic colonizing and
noncolonizing strains of C. jejuni failed to distinguish the two
strains and revealed only one copy of the gene. Post-translational
modification may be an alternate explanation for the antigenic differences
seen between the two strains.
Received: 15 October 1996 / Accepted: 3 December 1996 |
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