嗜热菌
赫拉
劈理(地质)
生物传感器
磷酸化
DNA
检出限
适体
组合化学
化学
细胞生物学
生物物理学
分子生物学
生物
生物化学
细胞
色谱法
基因
古生物学
大肠杆菌
断裂(地质)
作者
Yuqing Qin,Weikang Ke,Yanan Zhou,Dingliang Zhu,Yingjun Li,Yonggang Hu
标识
DOI:10.1016/j.snb.2023.133753
摘要
As a next-generation biosensing technology, the application of Argonaute for analytical science research has attracted considerable attention. A Thermus thermophilus Argonaute (TtAgo)-based sensor is presented for detecting T4 polynucleotide kinase (PNK) activity considering that TtAgo’s multiple-turnover catalytic activity can be activated by binding to a 5′-phosphate termini of single-stranded DNA (ssDNA) for the cleavage of complementary strand. PNK catalyzes the phosphorylation of 5′-hydroxyl termini of ssDNA, and the phosphorylation product termed guide can activate the TtAgo-mediated complementary ssDNA reporter cleavage to release fluorescence signal. For the PNK activity monitoring, the developed method shows good analytic performance with a linear range of 0.0010–2 U mL−1 and a detection limit of 0.0010 U mL−1 without the use of a preamplification reaction. This method is successfully applied for the analysis of HeLa cells at a single-cell level. The developed method shows several advantages, including high sensitivity, low cost, facile operation, and has remarkable potential application for early disease diagnosis, drug screening, and clinical treatment.
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