Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase
Reexamination Certificate
2007-01-24
2008-10-21
Gitomer, Ralph (Department: 1657)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving hydrolase
C435S006120, C435S199000, C436S024000
Reexamination Certificate
active
07439035
ABSTRACT:
The invention relates to a substrate and method for the assay of ribonuclease H activity. The substrate comprises a DNA/RNA heteroduplex molecule having fluorophore and quencher molecules in close proximity, which emit a background level of fluorescence in the absence of ribonuclease H activity due to quenching of the fluorophore as a result of fluorescence resonance energy transfer. Ribonuclease H cleavage of the heteroduplex results in an increase in fluorescence emission due to the destabilization and separation of the DNA and RNA molecules and their associated fluorophore and quencher molecules. The assay method of the present invention is suitable for use in screening (e.g. high throughput screening) for modulators, e.g. inhibitors, of the ribonuclease H activity associated with, for example, reverse transcriptase and ribonuclease H enzymes.
REFERENCES:
patent: 6268146 (2001-07-01), Shultz et al.
patent: 6287765 (2001-09-01), Cubicciotti
patent: 6573045 (2003-06-01), Karn et al.
patent: 6773885 (2004-08-01), Walder et al.
Min Kyung-Lyum
Parniak Michael A.
Gitomer Ralph
McGill University
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