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Forward Genetic Approaches in Chlamydia trachomatis
09:03

Forward Genetic Approaches in Chlamydia trachomatis

Published on: October 23, 2013

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The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum

Yibing Wang1, Lesley T Cutcliffe, Rachel J Skilton

  • 1Molecular Microbiology Group, Faculty of Medicine, University of Southampton, Southampton General Hospital, Southampton, UK.

Pathogens and Disease
|April 5, 2014
PubMed

Insights

Chlamydia species plasmids have a species-specific barrier. Researchers identified that a specific region, CDS 2, on the Chlamydia trachomatis plasmid is the main determinant of this plasmid tropism barrier.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Genetic transformation of Chlamydia trachomatis has been established using its endogenous plasmid.
  • A species-specific barrier prevents transformation of Chlamydia muridarum by the Chlamydia trachomatis plasmid.

Purpose of the Study:

  • To identify the specific regions of the Chlamydia trachomatis plasmid responsible for species-specific transformation barriers.
  • To understand the molecular basis for plasmid tropism between different Chlamydia species.

Main Methods:

  • Utilized forced recombination between the Chlamydia trachomatis plasmid (pGFP::SW2) and the Chlamydia muridarum plasmid (pNigg) in wild-type C. muridarum.
  • Selected for antibiotic-resistant transformants expressing green fluorescent protein.
  • Analyzed recovered plasmids to identify recombination breakpoints and exchanged regions.

Main Results:

  • Recombination occurred, with a ~1 kbp region of pGFP::SW2, including CDS 2, being exchanged with pNigg.
  • The resulting recombinant plasmid (pSW2NiggCDS2) was stably maintained in plasmid-cured C. muridarum under antibiotic selection.
  • The ability to select for recombinants indicated the barrier lies in plasmid replication or maintenance, not initial transformation.

Conclusions:

  • CDS 2, along with adjacent sequences, is the primary determinant of plasmid tropism in Chlamydia.
  • This finding clarifies the species-specific replication and maintenance mechanisms of Chlamydia plasmids.
Keywords:
Chlamydiaplasmidreplicationtransformationtropism

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