Bright stable luminescent yeast using bacterial luciferase as a sensor.
Bright luminescent yeast cells with light intensities similar to bacteria containing luciferase (LuxAB) were generated by providing saturating nontoxic levels of the substrates for the bioluminescence reaction (FMNH(2)+O(2) and fatty aldehyde-->light). Z-9-Tetradecenal added to yeast (+luxAB) gave a luminescent signal close to that with decanal with the signal remaining strong for >24h while luminescence of yeast with decanal decayed to less than 0.01% of that with Z-9-tetradecenal after 2min. Moreover, yeast survived in 0.5% (v/v) Z-9-tetradecenal while 0.005% (v/v) decanal was lethal. Luminescence of yeast (+luxAB) was also stimulated 100-fold by transformation with the NADPH-specific FMN reductase (FRP) from Vibrio harveyi. The recognition of the nontoxicity and high luminescence generated by Z-9-tetradecenal and the generation of high levels of FMNH(2) in yeast by transformation with a flavin reductase provide evidence for the strong potential use of bacterial luciferase as the light-emitting sensor of choice in eukaryotic organisms.
Szittner
Rose
R
Department of Biochemistry, McGill University, Montreal, PQ, Canada.
Jansen
Gregor
G
Thomas
David Y
DY
Meighen
Edward
E
D000447:Aldehydes / Q000494:pharmacology; D002455:Cell Division; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D005415:Flavins / Q000378:metabolism; D008027:Light; D008156:Luciferases / Q000737:chemistry* / Q000378:metabolism; D008163:Luminescent Measurements; D010088:Oxidoreductases / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013997:Time Factors
D000447:Aldehydes; D005415:Flavins; D010088:Oxidoreductases; D008156:Luciferases