Highly Effective Colorimetric and Visual Detection of ATP by a DNAzyme-Aptamer Sensor
Highly Effective Colorimetric and Visual Detection of ATP by a DNAzyme-Aptamer Sensor
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DOI：
10.1002/cbdv.201000130
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发表时间：
2011-03-01
影响因子：
2.9
通讯作者：
Deng, Le
中科院分区：
文献类型：
--
作者：
Liu, Fang;Zhang, Juan;Deng, Le
建立了一种利用脱氧核酶-核酸适体传感器检测三磷酸腺苷(ATP)的新方法。使用的脱氧核酶是一种单链DNA，可以与氯化血红素结合。该适配子是一种抗ATP适配子，是一种单一的短核酸序列，可以与许多靶点特异结合。设计了两条DNA序列：1)由抗ATP适配子(识别部分)和DNAzyme(信号转导部分)两部分组成的功能链(A链)；2)能与A链部分杂交的阻滞链(B链)。杂交链A和B是通过加入ATP和氯化血红素而展开的，阻滞链和功能链与ATP和氯化血红素的复合体在溶液中。功能链上的脱氧核酶与氯化高铁血红素形成G-四链体，然后催化H_2O_2氧化2，2‘-azinobis(3-ethylbenzthiazoline-6-sulfonic酸(ABTS(2-))生成有色ABTS(中心点-)自由基。肉眼可以清楚地观察到该反应引起的颜色变化，并在414 nm处记录了吸光度。检测下限为1×10(-6)m。
A new and simple method was developed to detect adenosine triphosphate (ATP) by using a DNAzyme-aptamer sensor. The DNAzyme used was a single-stranded DNA that could combine with hemin. The aptamer, a single, short nucleic acid sequence that can specifically bind with many targets, was an anti-ATP aptamer. Two DNA sequences were designed: i) a functional chain (Chain A) consisting of two parts, i. e., the anti-ATP aptamer (recognition part) and the DNAzyme (signal transduction part) and ii) a blocker chain (Chain B), which could partially hybridize with Chain A. The hybridized chains A and B were unfolded by the addition of ATP and hemin, and the blocker chain and the complex of the functional chain with ATP and hemin were in solution. The DNAzyme in the functional chain formed a G-quadruplex with hemin and then catalyzed the oxidation by H2O2 of 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS(2-)) to the colored ABTS(center dot-) radical. The color change caused by this reaction could be clearly observed by naked eye, and the absorbance was recorded at 414 nm. The detection limit was 1 x 10(-6) m.