1887

Abstract

Genetic variation due to mutation and phase variation has a considerable impact on the commensal and pathogenic behaviours of . In this study, we provide an example of how second-site mutations can interfere with gene function analysis in . Deletion of the flagellin B gene () in M1 resulted in mutant clones with inconsistent motility phenotypes. From the 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 mutant clones: a TA-dinucleotide deletion in and an A deletion in , respectively. Restoration of WT , using a newly developed genetic complementation system, confirmed that the second-site mutation caused the motility defect as opposed to the primary deletion of . 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.

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2015-12-01
2021-10-16
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