title: Ultrasensitive reporter protein detection in genetically engineered bacteria.
abstract: We demonstrate the use of laser-induced fluorescence confocal spectroscopy to measure analyte-stimulated enhanced green fluorescent protein (egfp) synthesis by genetically modified Escherichia coli bioreporter cells. Induction is measured in cell lysates and, since the spectroscopic focal volume is approximately the size of one bioreporter cell, also in individual live bacteria. This is, to our knowledge, the first ever proof-of-concept work utilizing instrumentation with single-molecule detection capability to monitor bioreporter response. Although we use arsenic inducible bioreporters here, the method is extensible to gfp/egfp bioreporters that are responsive to other substances.
authors.affiliation: Laboratory of Soil Science, Swiss Federal Institute of Technology (EPFL), CH-1015, Lausanne, Switzerland. mcwells@tntech.edu
authors.forename: Mona
authors.initials: M
authors.lastname: Wells
authors.forename: Michael
authors.initials: M
authors.lastname: Gösch
authors.forename: Rudolf
authors.initials: R
authors.lastname: Rigler
authors.forename: Hauke
authors.initials: H
authors.lastname: Harms
authors.forename: Theo
authors.initials: T
authors.lastname: Lasser
authors.forename: Jan Roelof
authors.initials: JR
authors.lastname: van der Meer
mesh_terms: D018053:Arsenites / Q000032:analysis*; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D030781:Organisms, Genetically Modified / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D018053:Arsenites; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C015001:arsenite