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Transformation of a plasmid-free, genital tract isolate of Chlamydia trachomatis with a plasmid vector carrying a deletion in CDS6 revealed that this gene regulates inclusion phenotype

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Author

  • Yibing Wang
  • Lesley T. Cutcliffe
  • Rachel J. Skilton
  • Kenneth Persson
  • Carina Bjartling
  • Ian N. Clarke

Summary, in English

The development of a plasmid-based genetic transformation protocol for Chlamydia trachomatis provides the basis for the detailed investigation of the function of the chlamydial plasmid and its individual genes or coding sequences (CDS). In this study we constructed a plasmid vector with CDS6 deleted (pCDS6KO) from the original Escherichia coli/C. trachomatis shuttle vector pGFP::SW2. pCDS6KO was transformed into a clinical isolate of C. trachomatis from Sweden that is plasmid-free (C. trachomatis SWFP-). Penicillin-resistant transformants expressing the green fluorescent protein were selected. These transformants did not stain with iodine, indicating that this property is regulated by CDS6 or its gene product. In addition, mature inclusions of C. trachomatis SWFP-transformed by pCDS6KO displayed an identical morphological phenotype to the untransformed plasmid-free recipient host. In this phenotype the morphology of inclusions was altered with the chlamydiae lining the periphery of the inclusion leaving a 'hole' in the centre. These green fluorescent inclusions appear 'doughnut-shaped' with an empty centre when examined under blue light, giving rise to a characteristic 'black hole' phenotype. Our study demonstrates the power of the new genetic system for investigating chlamydial gene function using gene deletion technology.

Publishing year

2013

Language

English

Pages

100-103

Publication/Series

Pathogens and Disease

Volume

67

Issue

2

Document type

Journal article

Publisher

Wiley-Blackwell

Topic

  • Microbiology in the medical area

Keywords

  • Chlamydia trachomatis
  • plasmid
  • transformation
  • deletion
  • CDS 6
  • glycogen biosynthesis

Status

Published

Research group

  • Clinical Microbiology, Malmö
  • Urogynaecology and Reproductive Pharmacology

ISBN/ISSN/Other

  • ISSN: 2049-632X