Label-free simple fluorescence assay of DNA polymerase using the G-quadruplex structure. Issue 2 (17th December 2015)

• Record Type: Journal Article
• Authors: Kim, Seyeon; Gang, Jongback
• Is Part Of: Analytical methods. Volume 8:Issue 2(2016)
• Page Start: 275
• Page End: 279
• Publication Date: 2015-12-17
• Subjects: Chemistry, Analytic -- Periodicals; Analytical biochemistry -- Periodicals; Chemical laboratories -- Standards -- Periodicals; 543.1905
• DOI: 10.1039/c5ay02379c
• Languages: English
• ISSNs: 1759-9660

Abstract

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 meso porphyrin IX (NMM) which binds specifically to the G-quadruplex. 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 meso porphyrin 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.