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Issue 2, 2016
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Label-free simple fluorescence assay of DNA polymerase using the G-quadruplex structure

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Abstract

In this study, the G-quadruplex structure was used to detect the enzymatic activity and inhibition of the Klenow fragment exo (KF) in the presence of N-methyl mesoporphyrin IX (NMM) which binds specifically to the G-quadruplex. We found that KF synthesized complementary strands of the template DNA, leading to a decrease in fluorescence intensity in the presence of NMM due to the difficulty in G-quadruplex formation. Thus, KF activity was sensitively detected using NMM. In addition, daidzein, the major phytoestrogen in soy, inhibited DNA synthesis mediated by KF, and the half maximal inhibitory concentration (IC50) was determined to be 15.6 μM by nonlinear regression analysis.

Graphical abstract: Label-free simple fluorescence assay of DNA polymerase using the G-quadruplex structure

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Publication details

The article was received on 07 Sep 2015, accepted on 11 Oct 2015 and first published on 13 Oct 2015


Article type: Paper
DOI: 10.1039/C5AY02379C
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Anal. Methods, 2016,8, 275-279

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    Label-free simple fluorescence assay of DNA polymerase using the G-quadruplex structure

    S. Kim and J. Gang, Anal. Methods, 2016, 8, 275
    DOI: 10.1039/C5AY02379C

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