Using a deoxyribozyme ligase and rolling circle amplification to detect a non-nucleic acid analyte, ATP
Using a deoxyribozyme ligase and rolling circle amplification to detect a non-nucleic acid analyte, ATP
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DOI：
10.1021/ja043490u
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发表时间：
2005-02-23
影响因子：
15
通讯作者：
Ellington, AD
中科院分区：
文献类型：
--
作者：
Cho, EJ;Yang, LT;Ellington, AD
变构核酶（适体酶）已用于将小有机分析物（ATP）的结合转导至用于滚环扩增（RCA）的圆形模板的连接中。将 ATP 激活的脱氧核酶连接酶固定在载玻片上，并在添加 ATP 后催化圆形挂锁探针的连接。连接产物可以通过 RCA 直接扩增和可视化。所展示的偶联反应可以检测低至 1 μM 的 ATP，并且可以区分结构相似的核苷酸，例如 GTP、CTP 和 UTP。 RCA 忠实地模拟了脱氧核酶的 ATP 协同激活，产生了对 ATP 浓度做出响应的放大“开关”。
An allosteric ribozyme (aptazyme) has been used to transduce the binding of a small organic analyte (ATP) into the ligation of a circular template for rolling circle amplification (RCA). An ATP-activated deoxyribozyme ligase was immobilized on a glass slide and, upon addition of ATP, catalyzed the ligation of a circular padlock probe. The ligated products could be directly amplified and visualized via RCA. The coupled reaction exhibited could detect as little as 1 μM of ATP and could discriminate against structurally similar nucleotides such as GTP, CTP, and UTP. Cooperative ATP activation of the deoxyribozyme was faithfully mimicked by RCA, yielding an amplified “switch” that was responsive to ATP concentration.