Construction and Use of Flow Cytometry Optimized Plasmid-Sensor Strains: Genomes in Flux
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research
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 language | English |
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Title of host publication | Horizontal Gene Transfer |
Editors | Maria Boekels Gogarten, Johann Peter Gogarten, Lorraine C. Olendzenski |
Volume | 532 |
Publisher | Humana Press |
Publication date | 2009 |
Pages | 257-268 |
ISBN (Print) | 978-1-60327-852-2 |
ISBN (Electronic) | 978-1-60327-853-9 |
DOIs | |
Publication status | Published - 2009 |
Series | Methods in Molecular Biology |
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ISSN | 1064-3745 |
Bibliographical note
Keywords Plasmid stability - green fluorescence protein - flow cytometry - plasmid-sensor strain - lac repressor
ID: 11663620