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United States Department of Agriculture

Agricultural Research Service

Title: Subtractive Hybridization and 2-D Gel Electrophoresis Identifies Genomic Differences Between Campylobacter Jejuni Isolates Demonstrated to Differentially Colonize Chickens

Authors
item Hiett, Kelli
item Kuntz, Robin
item Seal, Bruce

Submitted to: Campylobacter Helicobacter and Related Organisms International Workshop
Publication Type: Abstract Only
Publication Acceptance Date: June 13, 2005
Publication Date: September 4, 2005
Citation: Hiett, K.L., Kuntz, R.L., Seal, B.S. 2005. Subtractive hybridization and 2-d gel electrophoresis identifies genomic differences between campylobacter jejuni isolates demonstrated to differentially colonize chickens. Campylobacter Helicobacter and Related Organisms International Workshop. C3, P. 49.

Technical Abstract: Campylobacter spp. are considered the leading bacterial etiology of acute gastroenteritis in humans. Evidence implicates poultry as a major source of the organism for human illness; however, the factors involved in colonization of poultry with Campylobacter spp. remain unclear. Determining colonization-associated factors at the genome level should facilitate our understanding of Campylobacter spp. contamination of poultry so that targeted intervention strategies can be developed. Suppressive subtractive hybridization was used to identify genomic differences between two Campylobacter jejuni isolates, 11168 and A74/C, that exhibit different colonization potentials in poultry. A total of three hundred and twenty-four clones resulting from the subtractions were analyzed. Two hundred and forty-eight (76.5%) of the clones were determined to contain inserts of approximately 300 to 600 base pairs in size. Preliminary DNA sequence analyses of cloned inserts revealed similarities to a putative glycosyltransferase from C. jejuni O:2 and O:36, to neuA (CMP-Neu Nac sythetase) from C. jeuni 0:23, and to a CA repeat region from Homo sapiens. Southern blot and Northern blot hybridization analyses of a panel of diverse Campylobacter jejuni strains were performed to determine the distribution and expression of differentially identified gene sequences. Additionally, 2-D gel electrophoresis was performed on both soluble and membrane protein extracts from the two isolates. Identification of different genes potentially involved in colonization of Campylobacter spp. in poultry will facilitate the development of targeted intervention strategies, thereby allowing the delivery of a safer product to consumers.

Last Modified: 12/27/2014
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