Direct and convenient measurement of plasmid stability in lab and clinical isolates of E. coli

Abstract Plasmids are important mobile elements in bacteria, contributing to evolution, virulence, and antibiotic resistance. Natural plasmids are generally large and maintained at low copy number and thus prone to be lost. Therefore, dedicated plasmid maintenance systems have evolved, leading to pl...

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Autores principales:	Siyi Chen, Mårten Larsson, Robert C. Robinson, Swaine L. Chen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c6caa5b12d5d4d2ea0f7888c17d52530
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spelling	oai:doaj.org-article:c6caa5b12d5d4d2ea0f7888c17d525302021-12-02T16:06:37ZDirect and convenient measurement of plasmid stability in lab and clinical isolates of E. coli10.1038/s41598-017-05219-x2045-2322https://doaj.org/article/c6caa5b12d5d4d2ea0f7888c17d525302017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05219-xhttps://doaj.org/toc/2045-2322Abstract Plasmids are important mobile elements in bacteria, contributing to evolution, virulence, and antibiotic resistance. Natural plasmids are generally large and maintained at low copy number and thus prone to be lost. Therefore, dedicated plasmid maintenance systems have evolved, leading to plasmid loss rates as low as 1 per 107 divisions. These low rates complicate studies of plasmid loss, as traditional techniques for measuring plasmid loss are laborious and not quantitative. To overcome these limitations, we leveraged a stringent negative selection system to develop a method for performing direct, quantitative measurements of plasmid loss in E. coli. We applied our method to gain mechanistic insights into a heterologously reconstituted segregation system in lab strains and clinical isolates of E. coli. We also performed direct stability studies of a currently circulating resistance plasmid in a clinical isolate, strain EC958, which is a member of the rapidly expanding global ST131 E. coli clone. Our results establish the foundational assays required to screen for small molecules targeting plasmid stability, which could complement current strategies for reducing the spread of antibiotic resistance, complementing other strategies for treating antibiotic resistant bacteria.Siyi ChenMårten LarssonRobert C. RobinsonSwaine L. ChenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Siyi Chen Mårten Larsson Robert C. Robinson Swaine L. Chen Direct and convenient measurement of plasmid stability in lab and clinical isolates of E. coli
description	Abstract Plasmids are important mobile elements in bacteria, contributing to evolution, virulence, and antibiotic resistance. Natural plasmids are generally large and maintained at low copy number and thus prone to be lost. Therefore, dedicated plasmid maintenance systems have evolved, leading to plasmid loss rates as low as 1 per 107 divisions. These low rates complicate studies of plasmid loss, as traditional techniques for measuring plasmid loss are laborious and not quantitative. To overcome these limitations, we leveraged a stringent negative selection system to develop a method for performing direct, quantitative measurements of plasmid loss in E. coli. We applied our method to gain mechanistic insights into a heterologously reconstituted segregation system in lab strains and clinical isolates of E. coli. We also performed direct stability studies of a currently circulating resistance plasmid in a clinical isolate, strain EC958, which is a member of the rapidly expanding global ST131 E. coli clone. Our results establish the foundational assays required to screen for small molecules targeting plasmid stability, which could complement current strategies for reducing the spread of antibiotic resistance, complementing other strategies for treating antibiotic resistant bacteria.
format	article
author	Siyi Chen Mårten Larsson Robert C. Robinson Swaine L. Chen
author_facet	Siyi Chen Mårten Larsson Robert C. Robinson Swaine L. Chen
author_sort	Siyi Chen
title	Direct and convenient measurement of plasmid stability in lab and clinical isolates of E. coli
title_short	Direct and convenient measurement of plasmid stability in lab and clinical isolates of E. coli
title_full	Direct and convenient measurement of plasmid stability in lab and clinical isolates of E. coli
title_fullStr	Direct and convenient measurement of plasmid stability in lab and clinical isolates of E. coli
title_full_unstemmed	Direct and convenient measurement of plasmid stability in lab and clinical isolates of E. coli
title_sort	direct and convenient measurement of plasmid stability in lab and clinical isolates of e. coli
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/c6caa5b12d5d4d2ea0f7888c17d52530
work_keys_str_mv	AT siyichen directandconvenientmeasurementofplasmidstabilityinlabandclinicalisolatesofecoli AT martenlarsson directandconvenientmeasurementofplasmidstabilityinlabandclinicalisolatesofecoli AT robertcrobinson directandconvenientmeasurementofplasmidstabilityinlabandclinicalisolatesofecoli AT swainelchen directandconvenientmeasurementofplasmidstabilityinlabandclinicalisolatesofecoli
_version_	1718384988688220160

Direct and convenient measurement of plasmid stability in lab and clinical isolates of E. coli

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