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

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

The development of genetic transformation technology for Chlamydia trachomatis using its endogenous plasmid has recently been described. Chlamydia muridarum cannot be transformed by the C. trachomatis plasmid, indicating a barrier between chlamydial species. To determine which regions of the plasmid...

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Main Authors: Wang, Yibing, Cutcliffe, Lesley T, Skilton, Rachel J, Ramsey, Kyle H, Thomson, Nicholas R, Clarke, Ian N
Format: Online
Language:English
Published: BlackWell Publishing Ltd 2014
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4314687/
id pubmed-4314687
recordtype oai_dc
spelling pubmed-43146872015-02-04 The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum Wang, Yibing Cutcliffe, Lesley T Skilton, Rachel J Ramsey, Kyle H Thomson, Nicholas R Clarke, Ian N Short Communication The development of genetic transformation technology for Chlamydia trachomatis using its endogenous plasmid has recently been described. Chlamydia muridarum cannot be transformed by the C. trachomatis plasmid, indicating a barrier between chlamydial species. To determine which regions of the plasmid conferred the species specificity, we used the novel approach of transforming wild-type C. muridarum carrying the endogenous plasmid pNigg and forced recombination with the C. trachomatis vector pGFP::SW2 which carries the complete C. trachomatis plasmid (pSW2). Penicillin and chloramphenicol-resistant transformants expressing the green fluorescent protein were selected. Recovery of plasmids from these transformants showed they were recombinants. The differences between the pSW2 and pNigg allowed identification of the recombination breakpoints and showed that pGFP::SW2 had exchanged a ∼ 1 kbp region with pNigg covering CDS 2. The recombinant plasmid (pSW2NiggCDS2) is maintained under antibiotic selection when transformed into plasmid-cured C. muridarum. The ability to select for recombinants in C. muridarum shows that the barrier is not at transformation, but at the level of plasmid replication or maintenance. Our studies show that CDS 2, together with adjoining sequences, is the main determinant of plasmid tropism. BlackWell Publishing Ltd 2014-10 2014-05-08 /pmc/articles/PMC4314687/ /pubmed/24700815 http://dx.doi.org/10.1111/2049-632X.12175 Text en © 2014 The Authors. Pathogens and Disease published by John Wiley & Sons on behalf of the Federation of European Microbiological Societies. http://creativecommons.org/licenses/by/3.0/ This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
author Wang, Yibing
Cutcliffe, Lesley T
Skilton, Rachel J
Ramsey, Kyle H
Thomson, Nicholas R
Clarke, Ian N
spellingShingle Wang, Yibing
Cutcliffe, Lesley T
Skilton, Rachel J
Ramsey, Kyle H
Thomson, Nicholas R
Clarke, Ian N
The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum
author_facet Wang, Yibing
Cutcliffe, Lesley T
Skilton, Rachel J
Ramsey, Kyle H
Thomson, Nicholas R
Clarke, Ian N
author_sort Wang, Yibing
title The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum
title_short The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum
title_full The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum
title_fullStr The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum
title_full_unstemmed The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum
title_sort genetic basis of plasmid tropism between chlamydia trachomatis and chlamydia muridarum
description The development of genetic transformation technology for Chlamydia trachomatis using its endogenous plasmid has recently been described. Chlamydia muridarum cannot be transformed by the C. trachomatis plasmid, indicating a barrier between chlamydial species. To determine which regions of the plasmid conferred the species specificity, we used the novel approach of transforming wild-type C. muridarum carrying the endogenous plasmid pNigg and forced recombination with the C. trachomatis vector pGFP::SW2 which carries the complete C. trachomatis plasmid (pSW2). Penicillin and chloramphenicol-resistant transformants expressing the green fluorescent protein were selected. Recovery of plasmids from these transformants showed they were recombinants. The differences between the pSW2 and pNigg allowed identification of the recombination breakpoints and showed that pGFP::SW2 had exchanged a ∼ 1 kbp region with pNigg covering CDS 2. The recombinant plasmid (pSW2NiggCDS2) is maintained under antibiotic selection when transformed into plasmid-cured C. muridarum. The ability to select for recombinants in C. muridarum shows that the barrier is not at transformation, but at the level of plasmid replication or maintenance. Our studies show that CDS 2, together with adjoining sequences, is the main determinant of plasmid tropism.
publisher BlackWell Publishing Ltd
publishDate 2014
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4314687/
_version_ 1613183444618051584

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