Enhancement in fluorescence response by a quencher for amiloride upon binding to thymine opposite an abasic site in a DNA duplex.
Enhancement in fluorescence response by a quencher for amiloride upon binding to thymine opposite an abasic site in a DNA duplex.
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DOI：
10.1016/j.aca.2010.06.042
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发表时间：
2010-08
影响因子：
6.2
通讯作者：
Yusuke Sato;Jing Tian;Toshiki Ichihashi;Y. Chinda;Zhiai Xu;Yuanfeng Pang;S. Nishizawa;N. Teramae
中科院分区：
文献类型：
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作者：
Yusuke Sato;Jing Tian;Toshiki Ichihashi;Y. Chinda;Zhiai Xu;Yuanfeng Pang;S. Nishizawa;N. Teramae
通过使用碘化物 (I−) 作为淬灭剂，我们成功改善了阿米洛利与 21 聚体 DNA 双链体中 AP 位点对面的胸腺嘧啶结合时的荧光响应。从荧光测量来看，与 NaCl 溶液相比，添加 NaI 作为猝灭剂以及盐来调节离子强度可有效抑制溶液中未结合的阿米洛利的背景荧光。 Stern-Volmer 分析表明，AP 位点与核碱基结合的阿米洛利未暴露于 NaI 猝灭剂。因此，获得了阿米洛利的高信号背景荧光响应。通过使用猝灭剂增强阿米洛利的荧光响应可以显着提高携带T靶碱基的DNA双链体的检测限。本研究提出的方法简单有效。本方法可适用于荧光团微环境在识别事件时发生变化的其他检测系统。
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.