In this section
@article{Mavridou:2016:10.1016/j.plasmid.2016.09.005,
author = {Mavridou, DAI and Gonzalez, D and Clements, A and Foster, KR},
doi = {10.1016/j.plasmid.2016.09.005},
journal = {Plasmid},
pages = {65--71},
title = {The pUltra plasmid series: a robust and flexible tool for fluorescent labeling of Enterobacteria},
url = {http://dx.doi.org/10.1016/j.plasmid.2016.09.005},
volume = {87-88},
year = {2016}
}
TY - JOUR
AB - Fluorescent labeling has been an invaluable tool for the study of living organisms andbacterial species are no exception to this. Here we present and characterize the pUltraplasmids which express constitutively a fluorescent protein gene (GFP, RFP, YFP or CFP)from a strong synthetic promoter and are suitable for the fluorescent labeling of a broad rangeof Enterobacteria. The amount of expressed fluorophore from these genetic constructs issuch, that the contours of the cells can be delineated on the basis of the fluorescent signalonly. In addition, labeling through the pUltra plasmids can be used successfully forfluorescence and confocal microscopy while unambiguous distinction of cells labeled withdifferent colors can be carried out efficiently by microscopy or flow cytometry. We comparethe labeling provided by the pUltra plasmids with that of another plasmid series encodingfluorescent proteins and we show that the pUltra constructs are vastly superior in signalintensity and discrimination power without having any detectable growth rate effects for thebacterial population. We also use the pUltra plasmids to produce mixtures of differentiallylabeled pathogenic Escherichia, Shigella and Salmonella species which we test duringinfection of mammalian cells. We find that even inside the host cell, different strains can bedistinguished effortlessly based on their fluorescence. We, therefore, conclude that the pUltraplasmids are a powerful labeling tool especially useful for complex biological experimentssuch as the visualization of ecosystems of different bacterial species or of enteric pathogensin contact with their hosts.
AU - Mavridou,DAI
AU - Gonzalez,D
AU - Clements,A
AU - Foster,KR
DO - 10.1016/j.plasmid.2016.09.005
EP - 71
PY - 2016///
SN - 1095-9890
SP - 65
TI - The pUltra plasmid series: a robust and flexible tool for fluorescent labeling of Enterobacteria
T2 - Plasmid
UR - http://dx.doi.org/10.1016/j.plasmid.2016.09.005
UR - http://hdl.handle.net/10044/1/41049
VL - 87-88
ER -
CBRB
Imperial College London
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