Transposon Mutagenesis in Chlamydia trachomatis Identifies CT339 as a ComEC Homolog Important for DNA Uptake and Lateral Gene Transfer

MBio
Scott D LaBrieP Scott Hefty

Abstract

Transposon mutagenesis is a widely applied and powerful genetic tool for the discovery of genes associated with selected phenotypes. Chlamydia trachomatis is a clinically significant, obligate intracellular bacterium for which many conventional genetic tools and capabilities have been developed only recently. This report describes the successful development and application of a Himar transposon mutagenesis system for generating single-insertion mutant clones of C. trachomatis This system was used to generate a pool of 105 transposon mutant clones that included insertions in genes encoding flavin adenine dinucleotide (FAD)-dependent monooxygenase (C. trachomatis148 [ct148]), deubiquitinase (ct868), and competence-associated (ct339) proteins. A subset of Tn mutant clones was evaluated for growth differences under cell culture conditions, revealing that most phenocopied the parental strain; however, some strains displayed subtle and yet significant differences in infectious progeny production and inclusion sizes. Bacterial burden studies in mice also supported the idea that a FAD-dependent monooxygenase (ct148) and a deubiquitinase (ct868) were important for these infections. The ct339 gene encodes a hypothetical protein with limi...Continue Reading

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Citations

Sep 11, 2019·Journal of Bacteriology·Robert J SuchlandDaniel D Rockey
Sep 11, 2019·Journal of Bacteriology·Yibing WangKevin Hybiske
Sep 9, 2020·FEMS Microbiology Reviews·Barbara S Sixt
Aug 8, 2020·Frontiers in Immunology·Gabrielle Keb, Kenneth A Fields
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May 1, 2021·Pathogens and Disease·Arkaprabha Banerjee, David E Nelson
Jul 20, 2021·Frontiers in Microbiology·Hanna MartiNicole Borel
Sep 14, 2021·Pathogens and Disease·Jason R Hunt, Jason A Carlyon

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Datasets Mentioned

BETA
ACUI01000000

Methods Mentioned

BETA
PCR
Confocal microscopy
ubiquitination
Genome Sequencing
PCRs

Software Mentioned

SVM
Geneious
MEMSAT
GraphPad Prism
TMHMM2
HMMTOP
BLAST
SPLIT
MIPAR
Savvy Scalable

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