Flavin mononucleotide-based fluorescent reporter proteins outperform green fluorescent protein-like proteins as quantitative in vivo real-time reporters.
Fluorescent proteins of the green fluorescent protein (GFP) family are commonly used as reporter proteins for quantitative analysis of complex biological processes in living microorganisms. Here we demonstrate that the fluorescence signal intensity of GFP-like proteins is affected under oxygen limitation and therefore does not reflect the amount of reporter protein in Escherichia coli batch cultures. Instead, flavin mononucleotide (FMN)-binding fluorescent proteins (FbFPs) are suitable for quantitative real-time in vivo assays under these conditions.
Drepper
Thomas
T
Institute of Molecular Enzyme Technology, Heinrich Heine University Duesseldorf, Research Center Juelich, Stetternicher Forst, D-52426 Juelich, Germany. t.drepper@fz-juelich.de
Huber
Robert
R
Heck
Achim
A
Circolone
Franco
F
Hillmer
Anne-Kathrin
AK
Büchs
Jochen
J
Jaeger
Karl-Erich
KE
D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005486:Flavin Mononucleotide / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D013997:Time Factors
D029968:Escherichia coli Proteins; D049452:Green Fluorescent Proteins; D005486:Flavin Mononucleotide; D010100:Oxygen