Open AccessQuantification of plasmid loss in Escherichia coli cells by use of flow cytometry
Author(s) -
Bahl Martin Iain,
S?rensen S?ren Johannes,
Hestbjerg Hansen Lars
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(04)00015-1
Subject(s) - plasmid , flow cytometry , escherichia coli , biology , microbiology and biotechnology , population , phenotype , enterobacteriaceae , gene , genetics , demography , sociology
A method was developed to study plasmid stability in Escherichia coli cells, which utilised the high speed analysis properties of flow cytometry. To discriminate between plasmid‐harbouring cells and plasmid‐free cells a plasmid‐encoded Lac repressor protein was used to regulate the expression of a chromosomally inserted green fluorescent protein gene in the host cells. Flow cytometric analysis enabled detection and quantification of plasmid‐free cells due to their green fluorescent phenotype. The reported system offers real‐time analysis in combination with a very low detection level of plasmid loss in bacterial populations. This could be useful in future investigations of plasmid stability and population selection in bacterial communities.