| 摘要 |
Detecting an enzymatically active substance (I) that causes sequence-specific depurination or depyrimidination of a nucleic acid (NA) without strand breakage or has sequence-specific endonuclease activity. Detecting an enzymatically active substance (I) that causes sequence-specific depurination or depyrimidination of a nucleic acid (NA) without strand breakage or has sequence-specific endonuclease activity. The test sample is contacted with a substrate (II) that contains 3 NA segments: A and C that are complementary and hybridize under physiological conditions, forming a hairpin structure, and an intermediate segment B, and B (or A and C after hybridization) contains a recognition motif for the activity being determined. (II) also includes a fluorophore/quencher (F/Q) pair with one component linked to A and the other to C, so that when these segments are hybridized, F and Q are sufficiently close to each other for quenching to occur. Where (I) is being detected, the sample is also contacted with an agent (III) that cuts B specifically at the site where loss of a base has occurred. Any light emitted from F is then detected specifically. An independent claim is also included for a kit comprising (II) and (III). |
