Use of vitamin B2 for fluorescence detection of thymidine-related single-nucleotide polymorphisms

Seiichi Nishizawa, N. B. Sankaran, Takehiro Seino, Ying Yu Cui, Qing Dai, Chun Yan Xu, Keitaro Yoshimoto, Norio Teramae

Research output: Contribution to journalArticlepeer-review

25 Citations (Scopus)

Abstract

In combination with abasic site (AP site)-containing DNAs, potential use of a biotic fluorescence compound, Vitamin B2 (riboflavin), is demonstrated for the fluorescence detection of the thymine (T)-related single-nucleotide polymorphisms. Our method is based on construction of the AP site in DNA duplexes, which allows small ligands to bind to target nucleotides accompanied by fluorescence signaling: an AP site-containing probe DNA is hybridized with a target DNA so as to place the AP site toward a target nucleobase, by which hydrophobic microenvironments are provided for ligands to recognize target nucleotides through stacking and hydrogen-bonding interactions. In 10 mM sodium cacodylate buffer solutions (pH 7.0) containing 100 mM NaCl and 1.0 mM EDTA, Vitamin B2 is found to selectively bind to T (K 11 = 1.8 × 106 M-1 at 5°C) over other nucleobases, and this is accompanied by significant quenching of its fluorescence. While the sensing functions depend on the flanking sequences to the AP site, Vitamin B2 is applicable to the detection of T/C (cytosine), T/G (guanine) and T/A (adenine) mutation sequences of the CYP2A6 gene, where the flanking nucleobases are guanines in both positions (-GXG-, X = AP site).

Original languageEnglish
Pages (from-to)133-139
Number of pages7
JournalAnalytica Chimica Acta
Volume556
Issue number1
DOIs
Publication statusPublished - 2006 Jan 18

Keywords

  • Abasic site
  • Fluorescence detection
  • Hydrogen bond
  • Ligand
  • Single-nucleotide polymorphism
  • Stacking
  • Vitamin B

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

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