TY - JOUR
T1 - Enhancement in fluorescence response by a quencher for amiloride upon binding to thymine opposite an abasic site in a DNA duplex
AU - Sato, Yusuke
AU - Tian, Jing
AU - Ichihashi, Toshiki
AU - Chinda, Yuka
AU - Xu, Zhiai
AU - Pang, Yuanfeng
AU - Nishizawa, Seiichi
AU - Teramae, Norio
PY - 2010/8/1
Y1 - 2010/8/1
N2 - By using iodide (I-) as a quencher, we successfully improve the fluorescence response of amiloride when binding to thymine opposite an AP site in a 21-meric DNA duplex. From fluorescence measurements, as compared to the NaCl solutions, the addition of NaI as a quencher as well as salt to adjust the ionic strength effectively suppresses the background fluorescence from unbound amiloride in a solution. The Stern-Volmer analysis shows that the bound amiloride to the nucleobase at the AP site is unexposed to NaI quencher. Therefore the high signal-to-background fluorescence response of amiloride is obtained. Such enhancement in fluorescence response of amiloride by using the quencher can provide the significant improvement of the detection limit for DNA duplexes carrying T target base. The method presented in this study is simple and effective. The present method could be applicable to other detection system where microenvironment of fluorophores changes at a recognition event.
AB - By using iodide (I-) as a quencher, we successfully improve the fluorescence response of amiloride when binding to thymine opposite an AP site in a 21-meric DNA duplex. From fluorescence measurements, as compared to the NaCl solutions, the addition of NaI as a quencher as well as salt to adjust the ionic strength effectively suppresses the background fluorescence from unbound amiloride in a solution. The Stern-Volmer analysis shows that the bound amiloride to the nucleobase at the AP site is unexposed to NaI quencher. Therefore the high signal-to-background fluorescence response of amiloride is obtained. Such enhancement in fluorescence response of amiloride by using the quencher can provide the significant improvement of the detection limit for DNA duplexes carrying T target base. The method presented in this study is simple and effective. The present method could be applicable to other detection system where microenvironment of fluorophores changes at a recognition event.
KW - Abasic site
KW - DNA-binding ligand
KW - Fluorescence
KW - Quencher
KW - Single-nucleotide mutation
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U2 - 10.1016/j.aca.2010.06.042
DO - 10.1016/j.aca.2010.06.042
M3 - Article
C2 - 20708115
AN - SCOPUS:77955560434
VL - 675
SP - 49
EP - 52
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
IS - 1
ER -