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 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.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Bahl, M.I., Oregaard, G., Sørensen, S.J., Hansen, L.H. (2009). Construction and Use of Flow Cytometry Optimized Plasmid-Sensor Strains. In: Gogarten, M.B., Gogarten, J.P., Olendzenski, L.C. (eds) Horizontal Gene Transfer. Methods in Molecular Biology, vol 532. Humana Press. https://doi.org/10.1007/978-1-60327-853-9_15
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DOI: https://doi.org/10.1007/978-1-60327-853-9_15
Publisher Name: Humana Press
Print ISBN: 978-1-60327-852-2
Online ISBN: 978-1-60327-853-9
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