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dc.contributor.authorde Vries, Stefanen
dc.contributor.authorGupta, Srishtien
dc.contributor.authorBaig, Abiyaden
dc.contributor.authorL'Heureux, Joannaen
dc.contributor.authorPont, Elsaen
dc.contributor.authorWolanska, Dominika Pen
dc.contributor.authorMaskell, Duncanen
dc.contributor.authorGrant, Andrewen
dc.date.accessioned2015-09-21T12:04:23Z
dc.date.available2015-09-21T12:04:23Z
dc.date.issued2015-12-01en
dc.identifier.citationMicrobiology 2015, 161: 2316-2327. doi:10.1099/mic.0.000184.en
dc.identifier.issn1350-0872
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/251087
dc.description.abstractGenetic variation due to mutation and phase variation has a considerable impact on the commensal and pathogenic behaviours of Campylobacter jejuni. In this study, we provide an example of how second-site mutations can interfere with gene function analysis in C. jejuni. Deletion of the flagellin B gene (flaB) in C. jejuni M1 resulted in mutant clones with inconsistent motility phenotypes. From the flaB mutant clones picked for further analysis, two were motile, one showed intermediate motility and two displayed severely attenuated motility. To determine the molecular basis of this differential motility, a genome resequencing approach was used. Second-site mutations were identified in the severely attenuated and intermediate motility flaB mutant clones: a TA-dinucleotide deletion in fliW and an A deletion in flgD, respectively. Restoration of WT fliW, using a newly developed genetic complementation system, confirmed that the second-site fliW mutation caused the motility defect as opposed to the primary deletion of flaB. This study highlights the importance of (i) screening multiple defined gene deletion mutant clones, (ii) genetic complementation of the gene deletion and ideally (iii) screening for second-site mutations that might interfere with the pathways/mechanisms under study.
dc.description.sponsorshipThis work was funded by BBSRC grant RG66581.
dc.languageengen
dc.language.isoenen
dc.publisherMicrobiology Society
dc.rightsCreative Commons Attribution 4.0 International License
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectBacterial Proteinsen
dc.subjectCampylobacter jejunien
dc.subjectGene Deletionen
dc.subjectGene Expression Regulation, Bacterialen
dc.subjectSequence Deletionen
dc.titleMotility defects in Campylobacter jejuni defined gene deletion mutants caused by second-site mutations.en
dc.title.alternativeSpontaneous mutations in Campylobacter jejuni strain M1: A cautionary taleen
dc.title.alternativeSecond-site mutation causing motility defectsen
dc.typeArticle
prism.endingPage2327
prism.issueIdentifier12en
prism.publicationDate2015en
prism.publicationNameMicrobiologyen
prism.startingPage2316
prism.volume161en
dc.rioxxterms.funderBBSRC
dc.rioxxterms.projectidRG66581
dcterms.dateAccepted2015-09-17en
rioxxterms.versionofrecord10.1099/mic.0.000184en
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2015-12-01en
dc.contributor.orcidde Vries, Stefan [0000-0002-0823-208X]
dc.contributor.orcidMaskell, Duncan [0000-0002-5065-653X]
dc.contributor.orcidGrant, Andrew [0000-0001-9746-2989]
dc.identifier.eissn1465-2080
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idBBSRC (BB/K004514/1)
cam.issuedOnline2015-12-01en


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Except where otherwise noted, this item's licence is described as Creative Commons Attribution 4.0 International License