title: Label-free fluorescence turn-on detection of uracil DNA glycosylase activity based on G-quadruplex formation.
abstract: 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 0.01U/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.
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China; State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410081, China. Electronic address: macb2012@csu.edu.cn.
authors: Changbei
authors: C
authors: Ma
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors: Kefeng
authors: K
authors: Wu
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors: Haisheng
authors: H
authors: Liu
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors: Kun
authors: K
authors: Xia
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410081, China.
authors: Kemin
authors: K
authors: Wang
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China. Electronic address: junwang@csu.edu.cn.
authors: Jun
authors: J
authors: Wang
mesh_terms: D052160:Benzothiazoles; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D005472:Fluorouracil / Q000494:pharmacology; D054856:G-Quadruplexes; D006367:HeLa Cells; D006801:Humans; D013844:Thiazoles / Q000737:chemistry; D051981:Uracil-DNA Glycosidase / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D052160:Benzothiazoles; D005456:Fluorescent Dyes; D013844:Thiazoles; C009462:thioflavin T; D051981:Uracil-DNA Glycosidase; D005472:Fluorouracil
keywords: G-quadruplex; Label-free; Thioflavin T; Uracil DNA glycosylase