Title of article :
Use of vitamin B2 for fluorescence detection of thymidine-related single-nucleotide polymorphisms Original Research Article
Author/Authors :
Seiichi Nishizawa، نويسنده , , N.B. Sankaran، نويسنده , , Takehiro Seino، نويسنده , , Ying-Yu Cui، نويسنده , , Qing Dai، نويسنده , , Chunyan Xu، نويسنده , , Keitaro Yoshimoto، نويسنده , , Norio Teramae، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2006
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 (K11 = 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).
Keywords :
Single-nucleotide polymorphism , Vitamin B2 , Abasic site , Ligand , Hydrogen bond , Stacking , Fluorescence detection
Journal title :
Analytica Chimica Acta
Journal title :
Analytica Chimica Acta