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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 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 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.
1-5
Wang, Yibing
5561c404-84de-4032-afcd-cc7fd7282ad9
Cutcliffe, Lesley T.
88f242f4-b4f7-46d4-a1dd-c187dd60a773
Skilton, Rachel J.
74a38e2c-d4fb-4ebe-b560-3d335a76feef
Ramsey, Kyle H.
b93ffc54-69c6-4bf9-bfe5-cc078ed878fe
Thomson, Nicholas R.
5497a110-069d-4156-bccb-c77db572b1c2
Clarke, Ian N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b
Wang, Yibing
5561c404-84de-4032-afcd-cc7fd7282ad9
Cutcliffe, Lesley T.
88f242f4-b4f7-46d4-a1dd-c187dd60a773
Skilton, Rachel J.
74a38e2c-d4fb-4ebe-b560-3d335a76feef
Ramsey, Kyle H.
b93ffc54-69c6-4bf9-bfe5-cc078ed878fe
Thomson, Nicholas R.
5497a110-069d-4156-bccb-c77db572b1c2
Clarke, Ian N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b

Wang, Yibing, Cutcliffe, Lesley T., Skilton, Rachel J., Ramsey, Kyle H., Thomson, Nicholas R. and Clarke, Ian N. (2014) The genetic basis of plasmid tropism between Chlamydia trachomatis and Chlamydia muridarum. Pathogens and Disease, 1-5. (doi:10.1111/2049-632X.12175). (PMID:24700815)

Record type: Article

Abstract

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.

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More information

e-pub ahead of print date: 3 April 2014
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 365677
URI: https://eprints.soton.ac.uk/id/eprint/365677
PURE UUID: efcf8bb6-c23c-4d10-9187-430217d8b07d
ORCID for Ian N. Clarke: ORCID iD orcid.org/0000-0002-4938-1620

Catalogue record

Date deposited: 13 Jun 2014 15:23
Last modified: 06 Jun 2018 13:19

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