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dc.contributor.authorHartley-Tassell, Laurenen_US
dc.contributor.authorKaakoush, Nadeemen_US
dc.contributor.authorFord, Justinen_US
dc.contributor.authorKorolik, Victoriaen_US
dc.contributor.authorMendz, Georgeen_US
dc.date.accessioned2017-05-03T11:42:31Z
dc.date.available2017-05-03T11:42:31Z
dc.date.issued2009en_US
dc.date.modified2010-06-23T05:23:41Z
dc.identifier.issn17574749en_US
dc.identifier.doi10.1186/1757-4749-1-13en_AU
dc.identifier.urihttp://hdl.handle.net/10072/29905
dc.description.abstractPotential 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 DH5a 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.en_US
dc.description.peerreviewedYesen_US
dc.description.publicationstatusYesen_AU
dc.format.extent339114 bytes
dc.format.mimetypeapplication/pdf
dc.languageEnglishen_US
dc.language.isoen_AU
dc.publisherBioMed Centralen_US
dc.publisher.placeUnited Kingdomen_US
dc.relation.ispartofstudentpublicationYen_AU
dc.relation.ispartofpagefrom1en_US
dc.relation.ispartofpageto9en_US
dc.relation.ispartofjournalGut Pathogensen_US
dc.relation.ispartofvolume1en_US
dc.rights.retentionNen_AU
dc.subject.fieldofresearchBacteriologyen_US
dc.subject.fieldofresearchcode060501en_US
dc.titleCharacterisation of Campylobacter jejuni genes potentially involved in phosphonate degradationen_US
dc.typeJournal articleen_US
dc.type.descriptionC1 - Peer Reviewed (HERDC)en_US
dc.type.codeC - Journal Articlesen_US
dcterms.licensehttp://creativecommons.org/licenses/by/2.0en_US
gro.facultyOffice of the Snr Dep Vice Chancellor, Institute for Glycomicsen_US
gro.rights.copyrightCopyright 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.en_AU
gro.date.issued2009
gro.hasfulltextFull Text


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