分子信标
荧光
荧光团
核酸外切酶 III
DNA
化学
小分子
链霉亲和素
生物素
生物物理学
分子探针
核酸外切酶
生物传感器
杂交探针
生物素化
生物化学
寡核苷酸
生物
聚合酶
基因
物理
大肠杆菌
量子力学
作者
Qiang Liu,Xiaoqing Sun,Mei Liu,Yan Jin,Baoxin Li
标识
DOI:10.1016/j.snb.2020.127804
摘要
Small molecule-protein interaction plays a significant role in disease diagnosis and biomedical research. Most of the reported fluorescence strategies for small molecule-protein interaction still involve the use of costly fluorophore-labelled probes. In this work, a label-free fluorescent method for sensitive detection of small molecule-protein interaction is proposed. It is based on G-triplex‒based molecular beacon (G3-MB) and exonuclease III (Exo III)‒aided DNA recycling amplification. Biotin-streptavidin interaction is selected as one model for proof-of-concept. The biotin-linked double-strand DNA (dsDNA) is used as the binding probe. In the presence of streptavidin, Exo III hydrolyzes the binding probe to release the trigger DNA, and the released trigger DNA can hybridize G3-MB to trigger the Exo III cleavage process, accompanied by releasing the trigger DNA and G3 sequences. The trigger DNA can recycle, generating a large number of G3 sequences. The generated G3 sequences bind thioflavin T (ThT) to produce strong fluorescence emission. The interaction between small molecule–protein can be detected by the fluorescence reader. This method exhibits low detection limit (5.6 pg/mL), and owns the merits of the simplicity, high specificity and satisfactory applicability in serum sample.
科研通智能强力驱动
Strongly Powered by AbleSci AI