Construction and Use of Flow Cytometry Optimized Plasmid-Sensor Strains: Genomes in Flux

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearch

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 relies 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
EditorsMaria Boekels Gogarten, Johann Peter Gogarten, Lorraine C. Olendzenski
PublisherHumana Press
Publication date2009
ISBN (Print)978-1-60327-852-2
ISBN (Electronic)978-1-60327-853-9
Publication statusPublished - 2009
SeriesMethods in Molecular Biology

Bibliographical note

Keywords Plasmid stability - green fluorescence protein - flow cytometry - plasmid-sensor strain - lac repressor

ID: 11663620