Competitive binding of abasic site-binding ligands and masking ligands to DNA duplexes for the analysis of single-base Mutation

Yusuke Sato, Tomoe Kageyama, Seiichi Nishizawa, Norio Teramae

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

We describe a competitive binding assay for the analysis of single-base mutation by using abasic site (AP site)-specific binding fluorescent ligands and masking ligands bound to the AP site in DNA duplexes. 2-Amino-5,6,7-trimethyl-1,8-naphthyridine (ATMND) can strongly bind to not only cytosine (C), but also thymine (T) opposite an AP site in the DNA duplexes (K 11 /10 7 M -1 : C, 15; T, 7.0). By contrast, in the presence of lumichrome (Lch) as a masking ligand, ATMND shows a clear binding selectivity for C over T (K 11 /10 7 M -1 : C, 7.0; T, 0.57), which is ascribed to the effective masking effect of Lch to the ATMND-T interaction in the AP site-containing DNA duplexes. It was shown that the competitive binding of ATMND and Lch to the AP site allowed the highly selective detection of C-related single-base mutation in DNAs amplified by polymerase chain reaction.

Original languageEnglish
Pages (from-to)15-19
Number of pages5
Journalanalytical sciences
Volume29
Issue number1
DOIs
Publication statusPublished - 2013

ASJC Scopus subject areas

  • Analytical Chemistry

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