A "turn-on" and label-free fluorescent assay for the rapid detection of exonuclease III activity based on Tb3+-induced G-quadruplex conjugates

被引：15

作者：

Yang, WeiJuan ^{[1
]}

Ruan, YaJuan ^{[1
]}

Wu, WeiHua ^{[1
]}

Chen, PingPing ^{[1
]}

Xu, LiangJun ^{[1
]}

Fu, FengFu ^{[1
]}

机构：

[1] Fuzhou Univ, Dept Chem, Fujian Prov Key Lab Anal & Detect Food Safety, Minist Educ,Key Lab Anal & Detect Food Safety, Fuzhou 350108, Fujian, Peoples R China

来源：

ANALYTICAL AND BIOANALYTICAL CHEMISTRY | 2014年 / 406卷 / 18期

基金：

国家科技攻关计划;

关键词：

Nucleases; G-quadruplex; Exonuclease III; Fluorescent assay; Terbium; DNA; REGENERATION; DNAZYME;

D O I：

10.1007/s00216-014-7830-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A "turn-on" and label-free fluorescent assay for the specific, rapid, and sensitive detection of 3'aEuro parts per thousand a dagger'aEuro parts per thousand 5' exonuclease III activity is reported in this study. The assay is based on the Tb3+-promoted G-quadruplex, which lead to the enhancement of Tb3+ fluorescence due to the energy transfer from guanines. The proposed assay is highly simple, rapid, and cost-effective, and does not require sophisticated experimental techniques such as gel-based equipment or radioactive labels. It can be used for the rapid detection of exonuclease III activity with a detection limit of 0.8 U and a RSD (n = 6) < 5 %. Notably, no dye was covalently conjugated to the DNA strands, which offers the advantages of low-cost and being interference-free.

引用

页码：4535 / 4540

页数：6

共 23 条

[1] STRUCTURAL BASIS FOR THE 3'-5' EXONUCLEASE ACTIVITY OF ESCHERICHIA-COLI DNA-POLYMERASE-I - A 2 METAL-ION MECHANISM [J].