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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

Wang, Yibing ; Cutcliffe, Lesley T. ; Skilton, Rachel J. ; Persson, Kenneth LU ; Bjartling, Carina LU and Clarke, Ian N. (2013) In Pathogens and Disease 67(2). p.100-103
Abstract
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... (More)
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. (Less)
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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Chlamydia trachomatis, plasmid, transformation, deletion, CDS 6, glycogen biosynthesis
in
Pathogens and Disease
volume
67
issue
2
pages
100 - 103
publisher
Wiley-Blackwell
external identifiers
  • wos:000318348700003
  • scopus:84878131291
  • pmid:23620154
ISSN
2049-632X
DOI
10.1111/2049-632X.12024
language
English
LU publication?
yes
id
8f8cde2d-1754-4080-a2f5-0ce4bda14c8a (old id 3853339)
date added to LUP
2016-04-01 11:16:45
date last changed
2022-01-26 06:52:19
@article{8f8cde2d-1754-4080-a2f5-0ce4bda14c8a,
  abstract     = {{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.}},
  author       = {{Wang, Yibing and Cutcliffe, Lesley T. and Skilton, Rachel J. and Persson, Kenneth and Bjartling, Carina and Clarke, Ian N.}},
  issn         = {{2049-632X}},
  keywords     = {{Chlamydia trachomatis; plasmid; transformation; deletion; CDS 6; glycogen biosynthesis}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{100--103}},
  publisher    = {{Wiley-Blackwell}},
  series       = {{Pathogens and Disease}},
  title        = {{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}},
  url          = {{http://dx.doi.org/10.1111/2049-632X.12024}},
  doi          = {{10.1111/2049-632X.12024}},
  volume       = {{67}},
  year         = {{2013}},
}