Motility defects in Campylobacter jejuni defined gene deletion mutants caused by second-site mutations.
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Authors
Gupta, Srishti
Baig, Abiyad
L'Heureux, Joanna
Pont, Elsa
Wolanska, Dominika P
Publication Date
2015-12-01Alternative Title
Spontaneous mutations in Campylobacter jejuni strain M1: A cautionary tale
Second-site mutation causing motility defects
Journal Title
Microbiology
ISSN
1350-0872
Publisher
Microbiology Society
Volume
161
Issue
12
Pages
2316-2327
Language
English
Type
Article
Metadata
Show full item recordCitation
de Vries, S., Gupta, S., Baig, A., L'Heureux, J., Pont, E., Wolanska, D. P., Maskell, D., & et al. (2015). Motility defects in Campylobacter jejuni defined gene deletion mutants caused by second-site mutations.. Microbiology, 161 (12), 2316-2327. https://doi.org/10.1099/mic.0.000184
Abstract
Genetic 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.
Keywords
Bacterial Proteins, Campylobacter jejuni, Gene Deletion, Gene Expression Regulation, Bacterial, Sequence Deletion
Sponsorship
This work was funded by BBSRC grant RG66581.
Funder references
BBSRC (BB/K004514/1)
Identifiers
External DOI: https://doi.org/10.1099/mic.0.000184
This record's URL: https://www.repository.cam.ac.uk/handle/1810/251087
Rights
Creative Commons Attribution 4.0 International License
Licence URL: http://creativecommons.org/licenses/by/4.0/