Ultrasensitive reporter protein detection in genetically engineered bacteria.
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.
Wells
Mona
M
Laboratory of Soil Science, Swiss Federal Institute of Technology (EPFL), CH-1015, Lausanne, Switzerland. mcwells@tntech.edu
Gösch
Michael
M
Rigler
Rudolf
R
Harms
Hauke
H
Lasser
Theo
T
van der Meer
Jan Roelof
JR
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*
D018053:Arsenites; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C015001:arsenite