Characterisation of Campylobacter jejuni genes potentially involved in phosphonate degradation

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Title Characterisation of Campylobacter jejuni genes potentially involved in phosphonate degradation
Author Hartley, Lauren; Kaakoush, Nadeem O; Ford, Justin L; Korolik, Victoria; Mendz, George L
Journal Name Gut Pathogens
Year Published 2009
Place of publication United Kingdom
Publisher BioMed Central
Abstract Potential biological roles of the Campylobacter jejuni genes cj0641, cj0774c and cj1663 were investigated. The proteins encoded by these genes showed sequence similarities to the phosphonate utilisation PhnH, K and L gene products of Escherichia coli. The genes cj0641, cj0774c and cj1663 were amplified from the pathogenic C. jejuni strain 81116, sequenced, and cloned into pGEM-T Easy vectors. Recombinant plasmids were used to disrupt each one of the genes by inserting a kanamycin resistance (KmR) cassette employing site-directed mutagenesis or inverse PCR. Campylobacter jejuni 81116 isogenic mutants were generated by integration of the mutated genes into the genome of the wild-type strain. The C. jejuni mutants grew on primary isolation plates, but they could not be purified by subsequent passages owing to cell death. The mutant C. jejuni strains survived and proliferated in co-cultures with wild-type bacteria or in media in which wild-type C. jejuni had been previously grown. PCR analyses of mixed wild-type/mutant cultures served to verify the presence of the mutated gene in the genome of a fraction of the total bacterial population. The data suggested that each mutation inactivated a gene essential for survival. Rates of phosphonate catabolism in lysates of E. coli strain DH5α were determined using proton nuclear magnetic resonance spectroscopy. Whole-cell lysates of the wild-type degraded phosphonoacetate, phenylphosphonate and aminomethylphosphonate. Significant differences in the rates of phosphonate degradation were observed between lysates of wild-type E. coli, and of bacteria transformed with each one of the vectors carrying one of the C. jejuni genes, suggesting that these genes were involved in phosphonate catabolism.
Peer Reviewed Yes
Published Yes
Alternative URI http://dx.doi.org/10.1186/1757-4749-1-13
Copyright Statement Copyright 2009 Hartley et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Volume 1
Page from 1
Page to 9
ISSN 1757-4749
Date Accessioned 2010-01-27
Date Available 2010-06-23T05:23:41Z
Language en_AU
Research Centre Institute for Glycomics
Faculty Faculty of Science, Environment, Engineering and Technology
Subject Bacteriology
URI http://hdl.handle.net/10072/29905
Publication Type Journal Articles (Refereed Article)
Publication Type Code c1

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