Label-free fluorescence turn-on detection of uracil DNA glycosylase activity based on G-quadruplex formation

被引：17
作者：
Ma, Changbei [1 ,2 ,3 ]
Wu, Kefeng [1 ,2 ]
Liu, Haisheng [1 ,2 ]
Xia, Kun [1 ,2 ]
Wang, Kemin [3 ]
Wang, Jun [1 ,2 ]
机构：
[1] Cent S Univ, State Key Lab Med Genet, Changsha 410013, Hunan, Peoples R China
[2] Cent S Univ, Sch Life Sci, Changsha 410013, Hunan, Peoples R China
[3] Hunan Univ, State Key Lab Chemo Biosensing & Chemometr, Changsha 410081, Hunan, Peoples R China
来源：
TALANTA | 2016年 / 160卷
基金：
中国国家自然科学基金;
关键词：
Label-free; Uracil DNA glycosylase; Thioflavin T; G-quadruplex; REPAIR ENZYME-ACTIVITY; EXCISION-REPAIR; THIOFLAVIN T; SIGNAL AMPLIFICATION; PROBE; ASSAY; INDUCER; CANCER;
D O I：
10.1016/j.talanta.2016.07.048
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
We have developed a new methodology for fluorescence turn-on detection of uracil DNA glycosylase (UDG) activity based on G-quadruplex formation using a thioflavin T probe. In the presence of UDG, it catalyzed the hydrolysis of the uracil bases in the duplex DNA, resulting in the dissociation of the duplex DNA owing to their low melting temperature. Then, the probe DNA can be recognized quickly by the ThT dye and resulting in an increase in fluorescence. This approach is highly selective and sensitive with a detection limit of 0.01 U/mL. It is simple and cost effective without requirement of labeling with a fluorophore-quencher pair. This new method could be used to evaluate the inhibition effect of 5-fluorouracil on UDG activity, and become a useful tool in biomedical research. (C) 2016 Elsevier B.V. All rights reserved.
引用
收藏
页码：449 / 453
页数：5
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