Journal article
The Genetic Transformation of Chlamydia pneumoniae
mSphere, Vol.3(5), e00412-18
2018
Abstract
We demonstrate the genetic transformation of Chlamydia pneumoniae using a plasmid shuttle vector system which generates stable transformants. The equine C. pneumoniae N16 isolate harbors the 7.5-kb plasmid pCpnE1. We constructed the plasmid vector pRSGFPCAT-Cpn containing a pCpnE1 backbone, plus the red-shifted green fluorescent protein (RSGFP), as well as the chloramphenicol acetyltransferase (CAT) gene used for the selection of plasmid shuttle vector-bearing C. pneumoniae transformants. Using the pRSGFPCAT-Cpn plasmid construct, expression of RSGFP in koala isolate C. pneumoniae LPCoLN was demonstrated. Furthermore, we discovered that the human cardiovascular isolate C. pneumoniae CV-6 and the human community-acquired pneumonia-associated C. pneumoniae IOL-207 could also be transformed with pRSGFPCAT-Cpn. In previous studies, it was shown that Chlamydia spp. cannot be transformed when the plasmid shuttle vector is constructed from a different plasmid backbone to the homologous species. Accordingly, we confirmed that pRSGFPCAT-Cpn could not cross the species barrier in plasmidbearing and plasmid-free C. trachomatis, C. muridarum, C. caviae, C. pecorum, and C. abortus. However, contrary to our expectation, pRSGFPCAT-Cpn did transform C. felis. Furthermore, pRSGFPCAT-Cpn did not recombine with the wild-type plasmid of C. felis. Taken together, we provide for the first time an easy-to-handle transformation protocol for C. pneumoniae that results in stable transformants. In addition, the vector can cross the species barrier to C. felis, indicating the potential of horizontal pathogenic gene transfer via a plasmid.
Details
- Title
- The Genetic Transformation of Chlamydia pneumoniae
- Authors
- Kensuke Shima (Author) - University of Luebeck, GermanyMaximilian Wanker (Author) - University of Luebeck, GermanyRachel J Skilton (Author) - University of Southampton, United KingdomLesley T Cutcliffe (Author) - University of Southampton, United KingdomChristiane Schnee (Author) - Friedrich-Loeffler-lnstitute, GermanyThomas A Khol (Author) - Research Center Borstel, GermanyStefan Niemann (Author) - Research Center Borstel, GermanyJavier Geijo (Author) - University Vienna, AustriaMatthias Klinger (Author) - University of Luebeck, GermanyPeter Timms (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and EngineeringThomas Rattei (Author) - University Vienna, AustriaKonrad Sachse (Author) - FriedrichSchiller-Universität Jena, GermanyIan N Clarke (Author) - University of Southampton, United KingdomJan Rupp (Author) - University of Luebeck, Germany
- Publication details
- mSphere, Vol.3(5), e00412-18; 16
- Publisher
- American Society for Microbiology
- Date published
- 2018
- DOI
- 10.1128/mSphere.00412-18
- ISSN
- 2379-5042
- Copyright note
- Copyright © 2018 Shima et al. This is an openaccess article distributed under the terms of the Creative Commons Attribution 4.0 International license.
- Organisation Unit
- University of the Sunshine Coast, Queensland; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 99450674002621
- Output Type
- Journal article
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