title: Construction and use of flow cytometry optimized plasmid-sensor strains.
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.
authors.affiliation: Department of Microbiology, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Martin Iain
authors.initials: MI
authors.lastname: Bahl
authors.forename: Gunnar
authors.initials: G
authors.lastname: Oregaard
authors.forename: Søren J
authors.initials: SJ
authors.lastname: Sørensen
authors.forename: Lars Hestbjerg
authors.initials: LH
authors.lastname: Hansen
mesh_terms: D001419:Bacteria / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D020563:Gammaproteobacteria / Q000235:genetics; D042822:Genomic Instability; D049452:Green Fluorescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins