Intracellular Transposition, and Capture of Mobile Genetic Elements, Following Intercellular Conjugation of Multidrug Resistance Conjugative Plasmids from Clinical Enterobacteriaceae Isolates.

Tansirichaiya, Supathep, Goodman, Richard, Guo, Xinyu, Bulgasim, Issra, Samuelsen, Ørjan, Al-Haroni, Mohammed and Roberts, Adam ORCID: https://orcid.org/0000-0002-0760-3088 (2022) 'Intracellular Transposition, and Capture of Mobile Genetic Elements, Following Intercellular Conjugation of Multidrug Resistance Conjugative Plasmids from Clinical Enterobacteriaceae Isolates.'. Microbiology Spectrum, Vol 10, Issue 1, e02140-21.

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Abstract

Mobile genetic elements (MGEs) are often associated with antimicrobial resistance genes (ARGs). They are responsible for intracellular transposition between different replicons and intercellular conjugation, and are therefore important agents of ARG dissemination. Detection and characterisation of functional MGEs, especially in clinical isolates, would increase our understanding of the underlying pathways of transposition and recombination, and allow us to determine interventional strategies to interrupt this process.
Entrapment vectors can be used to capture active MGEs as they contain a positive selection genetic system conferring a selectable phenotype upon insertion of an MGE within certain regions of that system. Previously, we developed the pBACpAK entrapment vector that results in a tetracycline-resistant phenotype when MGEs translocate and disrupt the cI repressor gene. We have previously used pBACpAK to capture MGEs in clinical Escherichia coli isolates following transformation with pBACpAK.
In this study, we aimed to extend the utilisation of pBACpAK to other bacterial species. We utilised an MGE free recipient E. coli strain containing pBACpAK to capture MGEs on conjugative, ARG containing plasmids following conjugation from clinical Enterobacteriaceae donors.
Following the conjugative transfer of multiple conjugative plasmids, and screening for tetracycline resistance in these transconjugants, we captured several IS elements and novel transposons (Tn7350 and Tn7351), we detected the de novo formation of novel putative composite transposons where the pBACpAK located tet(A) is flanked by ISKpn25 from the transferred conjugative plasmid and we also detected the ISKpn14 mediated integration of an entire 119kb, blaNDM-1 containing conjugative plasmid from Klebsiella pneumoniae.
Importance
By analysing transposition activity within our MGE free recipient, we can gain insights into the interaction and evolution of multidrug resistance conferring MGEs following conjugation, including the movement of multiple ISs, the formation of composite transposons and the cointegration and, or recombination between different replicons in the same cell. This combination of recipient and entrapment vector will allow for fine-scale experimental studies into factors affecting intracellular transposition and MGE formation in, and from, ARG encoding MGEs from multiple species of clinically relevant Enterobacteriaceae.

Item Type:	Article
Subjects:	QU Biochemistry > Cells and Genetics > QU 300 General works QU Biochemistry > Genetics > QU 470 Genetic structures QU Biochemistry > Genetics > QU 475 Genetic processes QW Microbiology and Immunology > QW 45 Microbial drug resistance. General or not elsewhere classified.
Faculty: Department:	Biological Sciences > Department of Tropical Disease Biology
Digital Object Identifer (DOI):	https://doi.org/10.1128/spectrum.02140-21
Depositing User:	Cathy Waldron
Date Deposited:	08 Feb 2022 12:16
Last Modified:	08 Feb 2022 12:16
URI:	https://archive.lstmed.ac.uk/id/eprint/19750

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