DNA Detection through Signal Amplification by Using NADH:Flavin Oxidoreductase and Oligonucleotide-Flavin Conjugates as Cofactors

• Published in: Angewandte Chemie (International ed.) Vol. 44; no. 18; pp. 2764 - 2767
• Main Authors: Simon, Philippe, Dueymes, Cécile, Fontecave, Marc, Décout, Jean-Luc
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 29.04.2005; WILEY‐VCH Verlag
• Subjects: DNA - analysis; DNA - chemistry; DNA - genetics; DNA detection; DNA Probes - genetics; DNA Probes - metabolism; Escherichia coli; flavins; Flavins - chemistry; Flavins - metabolism; fluorescence; FMN Reductase - antagonists & inhibitors; FMN Reductase - metabolism; Molecular Sequence Data; Molecular Structure; Multienzyme Complexes - metabolism; NAD - metabolism; NADH, NADPH Oxidoreductases - metabolism; oligonucleotides; Oligonucleotides - chemistry; Oxidation-Reduction; oxidoreductases
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.200461145

Cofactors on the case: DNA detection by using an enzyme and its cofactor attached to an oligonucleotide is described. If the enzyme is functional only when the cofactor–oligonucleotide conjugate is not hybridized, substrate conversion monitored by fluorescence spectroscopy can be used as a reaction probe for hybridization (see scheme). Furthermore, as the enzyme turns over many copies of the substrate, the signal is greatly amplified.