用于3'→5'核酸外切酶活性检测的基于G-四链体的简单便捷的开启型荧光分析法。
Simple and convenient G-quadruplex-based turn-on fluorescence assay for 3' → 5' exonuclease activity.
作者信息
Leung Chung-Hang, Chan Daniel Shiu-Hin, Man Bradley Yat-Wah, Wang Chuan-Jen, Lam Wing, Cheng Yung-Chi, Fong Wang-Fun, Hsiao Wen-Luan Wendy, Ma Dik-Lung
出版信息
Anal Chem. 2011 Jan 15;83(2):463-6. doi: 10.1021/ac1025896. Epub 2010 Nov 29.
Abstract
A selective, oligonucleotide-based, label-free, turn-on fluorescence detection method for 3' → 5' exonuclease activity has been developed using crystal violet as a G-quadruplex-binding probe. The assay is highly simple and rapid, does not require the use of gel-based equipment or radioisotopic labeling, and is amenable to high-throughput and real-time detection. A proof-of-concept of this assay has been demonstrated for prokaryotic Exonuclease III (ExoIII) and human TREX1.
摘要
已开发出一种基于寡核苷酸的选择性、无标记、开启式荧光检测方法，用于检测3'→5'核酸外切酶活性，该方法使用结晶紫作为G-四链体结合探针。该检测方法非常简单快速，不需要使用基于凝胶的设备或放射性同位素标记，并且适用于高通量和实时检测。已针对原核核酸外切酶III（ExoIII）和人类TREX1证明了该检测方法的概念验证。
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