Label-free colorimetric assay for T4 polynucleotide kinase/phosphatase activity and its inhibitors based on G-quadruplex/hemin DNAzyme

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• 作者：Haisheng Liu^a ; Changbei Ma^a ; ^{macb2012@csu.edu.cn} ; Jun Wang^a ; Hanchun Chen^a ; Kemin Wang^b
• 关键词：Colorimetric assay ; T4 polynucleotide kinase/phosphatase ; G-quadruplex/hemin DNAzyme
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：15 January 2017
• 年：2017
• 卷：517
• 期：Complete
• 页码：18-21
• 全文大小：579 K
• 卷排序：517

文摘

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.