Inducible fluorescence assay

In one aspect, the invention provides fluorescent probes and assays. The probes include a fluorophore-quencher pair that undergoes a switch from dark to fluorescent in response to a reaction of the quencher. The switch of the probe from dark to fluorescent is typically mediated by an enzyme that act...

Ausführliche Beschreibung

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Bibliographische Detailangaben
Hauptverfasser:	COOK RONALD M, ARMSTRONG ELIANA S, JOHANSSON HANS E
Format:	Patent
Sprache:	eng
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES DERIVATIVES THEREOF ENZYMOLOGY FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES MEASURING MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MUTATION OR GENETIC ENGINEERING NUCLEIC ACIDS NUCLEOSIDES NUCLEOTIDES ORGANIC CHEMISTRY PHYSICS PROCESSES OF PREPARING SUCH COMPOSITIONS SPIRITS SUGARS TESTING VINEGAR WINE
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Beschreibung
Zusammenfassung:	In one aspect, the invention provides fluorescent probes and assays. The probes include a fluorophore-quencher pair that undergoes a switch from dark to fluorescent in response to a reaction of the quencher. The switch of the probe from dark to fluorescent is typically mediated by an enzyme that acts directly or indirectly on the quencher, interfering with its ability to quench fluorescence emission from the fluorophore. In another aspect, the invention provides a reporter gene assay system and methods of using this system. The assay system includes a fluorophore-quencher probe and an enzyme that acts directly or indirectly on the quencher, increasing the fluorescent emission of the fluorophore. In still other aspects, the invention provides nucleic acid constructs and cells expressing the peptide products of these constructs. In assays of the invention, the presence of a target substance is detected by the switching of fluorescence mediated by the change in oxidation state of the quencher. The assay systems are of use in numerous assay formats, e.g., confirming the expression of the enzyme that acts on the quencher, and/or detecting a species conjugated to the enzyme.