Bioluminescence detection system of mutagen using firefly luciferase genes introduced in Escherichia coli lysogenic strain.
A rapid and convenient microbial sensing system for mutagens was developed based upon the induction of prophage from Escherichia coli lysogenic strain and bioluminescence. The system consisted of lysogenic E. coli encoding firefly luciferase genes and a photodetection system. Measurement of mutagen mitomycin C was achieved by measuring the luminescence intensity emitted from E. coli lysogenic strain for the recombinant phage in the presence of luminescence substrates. Approximately 1 h after addition of mitomycin C, the luminescence began to be observed, and 3 h after, it attained a level of 2 times greater than that of 1 h. Irradiation with ultraviolet light also produced light based on induction of phage from the E. coli lysogenic strain for the recombinant phage. On the other hand, when nonmutagenic toxic compounds like sodium azide were added to the reaction medium, luminescence was not observed. Mitomycin C could be detected within 1 h with this sensing system, at concentrations down to 10(2) ng/assay.
Lee
S M
SM
Research Center for Advanced Science and Technology, University of Tokyo, Japan.
Suzuki
M
M
Kumagai
M
M
Ikeda
H
H
Tamiya
E
E
Karube
I
I
D010582:Bacteriophage lambda / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D008163:Luminescent Measurements; D008242:Lysogeny; D016685:Mitomycin / Q000032:analysis* / Q000494:pharmacology; D009152:Mutagenicity Tests; D009153:Mutagens / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet; D014775:Virus Activation
D009153:Mutagens; D016685:Mitomycin; D008156:Luciferases