Spectrofluorometry of Ions and Small Molecules Based on Conformational Changes of Specific Oligonucleotides with o-Phthalaldehyde-beta-mercaptoethanol
文献类型： 外文期刊
作者： Liao Qie-gen 1 ; Wang Jian 1 ; Long Yun-fei 1 ; Li Yuan-fang 1 ;
作者机构： 1.Southwest Univ, Coll Chem & Chem Engn, Key Lab Luminescence & Real Time Anal, Minist Educ, Chongqing 400715, Peoples R China
2.Jiangxi Acad Agr Sci, Key Lab Qual & Safety Agr Prod, Nanchang 330200, Peoples R China
关键词： Aptamer;o-Phthalaldehyde;beta-Mercaptoethanol;K+;denosine-5 '-triphosphoric acid
期刊名称：CHEMICAL RESEARCH IN CHINESE UNIVERSITIES （ 影响因子：1.307； 五年影响因子：0.979 ）
ISSN： 1005-9040
年卷期： 2010 年 26 卷 3 期
页码：
收录情况： SCI
摘要： Specific oligonucleotides such as telomere DNA and aptamer often undergo conformational changes upon ligand binding. Composite reagent composed of o-phthalaldehyde and beta-mercaptoethanol(OPAME) has been extensively applied to fluorescent detection of amino compounds based on the reaction of primary amino-group, herein we proposed a general spectrofluorometry for ions and small molecules due to conformational changes upon ligand binding taking K+ and ATP as examples. In a borate controlled buffer medium, telomere DNA could react with OPAME, giving a thio-subtituted isoindole compound with strong fluorescence emission at 455 nm when excited at 340 nm. It was found that however, the fluorescence emission was greatly reduced in the presence of K+ since the formation of the quadruplex structure inhibits the reaction activity of amino-groups of telomere DNA. In order to testify the general application of OPAME reagent based on the conformational change of oligonucleotides, we further proposed a sensitive method of ATP based on its highly selective interaction with ATP-aptamer. The above mentioned applications show that the spectrofluorometry with the aid of OPAME reagent is simple, label free that is expected to be potentially general for DNA conformational change-based target detection.
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