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用于三磷酸腺苷检测的银纳米簇分子信标的设计与合成

Design and Synthesis of Ag Nanocluster Molecular Beacon for Adenosine Triphosphate Detection.

作者信息

Li Xiaoshuang, Zhang Hao, Zhao Ying, Lian Lili, Wang Xiyue, Gao Wenxiu, Zhu Bo, Lou Dawei

机构信息

Department of Analytical Chemistry, Jilin Institute of Chemical Technology, 45 Chengde Street, Jilin 132022, China.

出版信息

J Anal Methods Chem. 2019 Oct 13;2019:2786156. doi: 10.1155/2019/2786156. eCollection 2019.

Abstract

This study presents a fluorescence method for detecting adenosine triphosphate (ATP) based on a label-free Ag nanocluster molecular beacon (MB) with high sensitivity. The sensor contains a hairpin-shaped MB, two short single-stranded DNA strands, and T4 DNA ligase. The MB consists of three parts, which are the template DNA sequence for synthesizing Ag nanoclusters at the 5' end, the middle DNA with a hairpin-shaped structure, and the guanine base-rich DNA sequence at the 3' end. The sensor exhibits high fluorescence intensity in the absence of ATP. However, when the probe is used for ATP detection, the two short DNA sequences in the sensor would form a long sequence by enzymatic ligation reaction; this long sequence opens the hairpin-shaped structure of the MB and decreases the fluorescence of the system. Under optimal analytical conditions, a clear linear relationship is observed between ATP concentration and fluorescence intensity in the range of 0.1-10 M. The interference presented by other small molecules during ATP detection is evaluated, and results confirm the good selectivity of the proposed sensor. Compared with traditional methods, the sensor is label free, easy to operate, inexpensive, and highly sensitive.

摘要

本研究提出了一种基于无标记银纳米簇分子信标(MB)的高灵敏度荧光法检测三磷酸腺苷(ATP)。该传感器包含一个发夹状的MB、两条短单链DNA和T4 DNA连接酶。MB由三部分组成,分别是5'端用于合成银纳米簇的模板DNA序列、具有发夹状结构的中间DNA以及3'端富含鸟嘌呤碱基的DNA序列。在没有ATP的情况下,该传感器表现出高荧光强度。然而,当该探针用于ATP检测时,传感器中的两条短DNA序列会通过酶促连接反应形成一个长序列;这个长序列会打开MB的发夹状结构并降低系统的荧光。在最佳分析条件下,在0.1 - 10 M范围内,ATP浓度与荧光强度之间呈现出明显的线性关系。评估了ATP检测过程中其他小分子的干扰情况,结果证实了所提出传感器具有良好的选择性。与传统方法相比,该传感器无标记、操作简便、成本低廉且灵敏度高。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ab9/6815610/d55860cfac6d/JAMC2019-2786156.001.jpg

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