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High throughput method for analysis of repeat number for 28 phase variable loci of C. jejuni strain NCTC11168

Lango-Scholey, Lea; Aidley, Jack; Woodacre, Alex; Jones, Michael A.; Bayliss, Christopher D.


Lea Lango-Scholey

Jack Aidley

Alex Woodacre

Christopher D. Bayliss


Mutations in simple sequence repeat tracts are a major mechanism of phase variation in several bacterial species including Campylobacter jejuni. Changes in repeat number of tracts located within the reading frame can produce a high frequency of reversible switches in gene expression between ON and OFF states. The genome of C. jejuni strain NCTC11168 contains 29 loci with polyG/polyC tracts of seven or more repeats. This protocol outlines a method for rapidly determining ON/OFF states of these 28 phase-variable loci in a large number of individual colonies. The method combines a series of multiplex PCR assays with a GeneScan assay and automated extraction of tract length, repeat number and expression state. This high throughput, multiplex assay has utility for detecting shifts in phase variation states within and between populations over time and for exploring the effects of phase variation on adaptation to differing selective pressures. An important output of this assay is combinatorial expression states that cannot be determined by other methods. This method can be adapted to analysis of phase variation in other C. jejuni strains and in a diverse range of bacterial species.


Lango-Scholey, L., Aidley, J., Woodacre, A., Jones, M. A., & Bayliss, C. D. (2016). High throughput method for analysis of repeat number for 28 phase variable loci of C. jejuni strain NCTC11168. PLoS ONE, 11(7), 1-15.

Journal Article Type Article
Acceptance Date Jul 5, 2016
Online Publication Date Jul 28, 2016
Publication Date Jul 28, 2016
Deposit Date Apr 5, 2017
Publicly Available Date Apr 5, 2017
Journal PLoS ONE
Electronic ISSN 1932-6203
Publisher Public Library of Science
Peer Reviewed Peer Reviewed
Volume 11
Issue 7
Article Number e0159634
Pages 1-15
Keywords Campylobacter, Phase variation, Colonisation, Genetics, Chickens
Public URL
Publisher URL
Additional Information Lango-Scholey L, Aidley J, Woodacre A, Jones MA, Bayliss CD (2016) High Throughput Method for Analysis of Repeat Number for 28 Phase Variable Loci of Campylobacter jejuni Strain
NCTC11168. PLoS ONE 11(7): e0159634.


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