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Direct and convenient measurement of plasmid stability in lab and clinical isolates of E-coli
Natl Univ Singapore, Yong Loo Lin Sch Med, Dept Med, 1E Kent Ridge Rd,NUHS Tower Block,Level 10, Singapore 119228, Singapore..
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology. ASTAR, Inst Mol & Cell Biol, 61 Biopolis Dr, Singapore 138673, Singapore.. (Genomik; Leif Andersson)
ASTAR, Inst Mol & Cell Biol, 61 Biopolis Dr, Singapore 138673, Singapore.;Natl Univ Singapore, Dept Biochem, Singapore 117597, Singapore.;Nanyang Technol Univ, NTU Inst Struct Biol, 59 Nanyang Dr, Singapore 636921, Singapore..
Natl Univ Singapore, Yong Loo Lin Sch Med, Dept Med, 1E Kent Ridge Rd,NUHS Tower Block,Level 10, Singapore 119228, Singapore.;Genome Inst Singapore, GERMS, 60 Biopolis St,Genome 02-01, Singapore 138672, Singapore.;Genome Inst Singapore, Infect Dis Grp, 60 Biopolis St,Genome 02-01, Singapore 138672, Singapore..
2017 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 7, 4788Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2017. Vol. 7, 4788
National Category
Basic Medicine
Identifiers
URN: urn:nbn:se:uu:diva-329997DOI: 10.1038/s41598-017-05219-xISI: 000404841100044PubMedID: 28684862OAI: oai:DiVA.org:uu-329997DiVA: diva2:1149048
Available from: 2017-10-13 Created: 2017-10-13 Last updated: 2017-10-13Bibliographically approved

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