Guanine-based chemiluminescence resonance energy transfer biosensing platform for the specific assay of uracil-DNA glycosylase activityElectronic supplementary information (ESI) available. See DOI: 10.1039/c6ay02964g
Assay of uracil-DNA glycosylase (UDG) activity is a crucial step for understanding some fundamental biochemical processes. Herein, we developed a simple and fast chemiluminescence (CL) method for the assay of UDG activity. In this method, one dsDNA probe containing fluorescein amidite-labeled G-rich...
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Zusammenfassung: | Assay of uracil-DNA glycosylase (UDG) activity is a crucial step for understanding some fundamental biochemical processes. Herein, we developed a simple and fast chemiluminescence (CL) method for the assay of UDG activity. In this method, one dsDNA probe containing fluorescein amidite-labeled G-rich ssDNA was used as a substrate for UDG. The dsDNA probe reacted with 3,4,5-trimethoxylphenylglyoxal to emit strong CL due to guanine-based chemiluminescence resonance energy transfer (CRET). The UDG-catalyzed uracil removal liberated G-rich ssDNA from the dsDNA substrate; then, in the presence of K
+
, G-rich ssDNA was converted to a G-quadruplex, which gives low CL emission. Therefore, using the CL response of the DNA probe as a signal indicator, UDG activity was determined in a simple process with high specificity, and a detection limit of 0.3 U mL
−1
was achieved without any signal amplification. This simple and rapid methodology may have potential application in the UDG-related clinical diagnoses and functional studies.
Guanine-based chemiluminescence resonance energy transfer system was used to detect uracil-DNA glycosylase (UDG) activity. The proposed method is simple and rapid. |
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ISSN: | 1759-9660 1759-9679 |
DOI: | 10.1039/c6ay02964g |