Novel tetracycline resistance gene tet(65) located on a multi-resistance Corynebacterium plasmid.

Kittl, Sonja; Brodard, Isabelle; Tresch, Milena; Perreten, Vincent (2024). Novel tetracycline resistance gene tet(65) located on a multi-resistance Corynebacterium plasmid. The journal of antimicrobial chemotherapy, 79(5), pp. 1023-1029. Oxford University Press 10.1093/jac/dkae066

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BACKGROUND

Corynebacterium (C.) sp. 22KM0430 related to C. oculi and isolated from a dog exhibited resistance to tetracycline, and its WGS analysis revealed a putative resistance gene on a 35 562-bp plasmid also harbouring the MLSB resistance gene erm(X).

OBJECTIVES

To characterize the novel tetracycline resistance gene tet(65) and demonstrate its functionality by expression in C. glutamicum and Escherichia coli and plasmid curing of the host strain.

METHODS

tet(65) was cloned with and without its repressor tetR(65) and expressed in C. glutamicum DSM20300 and E. coli DH5α. Plasmid was cured by non-selective passages. Minimal inhibitory concentrations (MICs) of tetracyclines were determined according to CLSI guidelines. Association of tet(65) with efflux was shown by the addition of reserpine to MIC assays. Phylogenetic position and transmembrane structure of Tet(65) were analysed using MEGA11 and DeepTMHMM.

RESULTS

Tet(65) shows 73% amino acid identity with the closest related Tet(Z), contains 12 transmembrane domains and is structurally related to the Major Facilitator Superfamily. The tetracycline MICs decreased in the plasmid-cured strain and increased when tet(65) was expressed in C. glutamicum and in E. coli. The MICs of tetracycline decreased in the presence of reserpine indicating that tet(65) functions as an efflux pump. A GenBank search also identified tet(65) in C. diphtheriae and Brevibacterium (B.) casei and B. luteolum.

CONCLUSIONS

A novel tetracycline efflux gene tet(65) was identified in a C. oculi related species and was also present in the human pathogen C. diphtheriae and in Brevibacterium species indicating broader potential for dissemination.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology > Molecular Bacterial Epidemiology and Infectiology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology

UniBE Contributor:

Kittl, Sonja Cornelia, Brodard, Isabelle, Tresch, Milena Laura, Perreten, Vincent

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 630 Agriculture

ISSN:

1460-2091

Publisher:

Oxford University Press

Language:

English

Submitter:

Pubmed Import

Date Deposited:

19 Mar 2024 07:58

Last Modified:

03 May 2024 09:41

Publisher DOI:

10.1093/jac/dkae066

PubMed ID:

38497972

BORIS DOI:

10.48350/194468

URI:

https://boris.unibe.ch/id/eprint/194468

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