Tet(C) gene transfer between Chlamydia suis strains occurs by homologous recombination after co-infection: Implications for spread of tetracycline-resistance among Chlamydiaceae

Children's Hospital Oakland Research Institute, UCSF Benioff Children's Hospital Oakland, Oakland, CA, 94609
Department of Infectious Diseases and Genetics, Emory University, Atlanta, GA, United States
DOI
10.7287/peerj.preprints.2489v1
Subject Areas
Cell Biology, Genomics, Microbiology, Molecular Biology, Infectious Diseases
Keywords
Chlamydia, homologous recombination, genomics, tetracycline resistance, tetC
Copyright
© 2016 Marti et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
Cite this article
Marti H, Kim H, Joseph S, Dojiri SS, Read T, Dean D. 2016. Tet(C) gene transfer between Chlamydia suis strains occurs by homologous recombination after co-infection: Implications for spread of tetracycline-resistance among Chlamydiaceae. PeerJ Preprints 4:e2489v1

Abstract

Chlamydia suis is a swine pathogen that also causes zoonotic infections. Many strains contain a tet(C) cassette conferring tetracycline resistance originally acquired by horizontal gene transfer from a Gram-negative donor. As we and others have described the capacity for Chlamydiaceae to exchange DNA by homologous recombination, tet(C)-containing C. suis strains represent a potential source for antibiotic-resistance spread within and between Chlamydia species. Here, we examine the genetics of tet(C)-transfer among C. suis strains. Tetracycline-sensitive C. suis strain S45 was simultaneously or sequentially co-infected with tetracycline-resistant C. suis strains in McCoy cells. Potential recombinants were clonally purified by plaque assay. C. suis strain Rogers132, which lacks IS200/IS605 transposases, was the most efficient donor, producing three confirmed recombinants of 56 (5.4%) clones with a minimal inhibitory concentration (MIC) of ~8µg/mL. Resistance was stable when recombinants were grown initially in tetracycline at twice the MIC of S45 (0.032 µg/mL). Genomic analysis revealed that tet(C) had integrated into the S45 genome by homologous recombination at two sites in different recombinants: a 55kb exchange between nrqF and pckG, and a 175kb exchange between kdsA and cysQ. Neither site was associated with repeats or motifs associated with recombination hotspots. Our findings show that cassette transfer into S45 has low frequency, does not require IS200/IS605 transposases, is stable if initially grown in tetracycline, and results in multiple genomic configurations. We provide a model for stable cassette transfer to better understand the capability for cassette acquisition by Chlamydia species that infect humans, a matter of public health importance.

Author Comment

This preprint submission to PeerJ Preprints was presented in part at the Eighth Meeting of the European Society for Chlamydia Research held in Oxford, England, September, 2016.

Supplemental Information

Supplementary Figures S1-S3

DOI: 10.7287/peerj.preprints.2489v1/supp-1

Supplementary Table S1: Primers used to identify putative recombinants

DOI: 10.7287/peerj.preprints.2489v1/supp-2

Supplementary Table S2. Database of 4,864 SNPs identified between the Rogers132 donor and S45 recipient

DOI: 10.7287/peerj.preprints.2489v1/supp-3

Pseudo contains of recombinant genome Rec3

DOI: 10.7287/peerj.preprints.2489v1/supp-4

Pseudo contains of recombinant genome Rec4

DOI: 10.7287/peerj.preprints.2489v1/supp-5

Pseudo contains of recombinant genome Rec5

DOI: 10.7287/peerj.preprints.2489v1/supp-6