Journal
ANALYTICAL BIOCHEMISTRY
Volume 517, Issue -, Pages 18-21Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2016.10.022
Keywords
Colorimetric assay; T4 polynucleotide kinase/phosphatase; G-quadruplex/hemin DNAzyme
Funding
- National Natural Science Foundation of China [21205142]
- Research Innovation Program for Graduates of Central South University [2016zzts580]
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Here we report a new approach for label-free colorimetric assay of T4 polynucleotide kinase/phosphatase (PNKP) activity based on G-quadruplex/hemin DNAzyme. In the presence of T4 PNKP, the DNA primer with a 3'-phosphate can be dephosphorylated into a 3'-hydroxyl and initiate a primer elongation reaction to open the hairpin probe, and leading to releasing the G-quartets. Under optimal conditions, the proposed method exhibited a considerable performance with a detection limit of 0.01 U/mL. Furthermore, the present assay can be used to study the potential T4 PNKP inhibitor screening, making it promise to be applied in the fields of drug discovery. (C) 2016 Elsevier Inc. All rights reserved.
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