Construction and Use of Flow Cytometry Optimized Plasmid-Sensor Strains

Martin Iain Bahl, Gunnar Oregaard, Søren J. Sørensen, Lars Hestbjerg Hansen

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

Abstract

Determining the stability of plasmids in bacterial populations is traditionally performed by isolating a large number of clones followed by screening for the presence of plasmids by replica transfer to plasmid-selective agar plates. This is often a laborious task, especially when the intrinsic stability of the plasmid is high. The method presented here relics on a phenotypic (green fluorescence protein) marker, which is switched on if the host bacteria loses the residing plasmid. The incorporation of flow cytometry for single-cell detection and discrimination between plasmid-free and plasmid-harboring cells in a bacterial population facilitates a very high throughput of cells and thus provides excellent sensitivity and statistics toward detecting even very low levels of plasmid instability.
Original languageEnglish
Title of host publicationHorizontal Gene Transfer
Volume532 / 3
PublisherHumana Press
Publication date2009
Pages257-268
ISBN (Print)9781603278522
Publication statusPublished - 2009
Externally publishedYes
SeriesMethods in Molecular Biology
ISSN1064-3745

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