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Linked bridge hybridizing-induced split G-quadruplex DNA machine and its application to uracil-DNA glycosylase detection.

Authors :
Zhang, Xiao Fang
Li, Na
Ling, Yu
Tang, Li
Li, Nian Bing
Luo, Hong Qun
Source :
Sensors & Actuators B: Chemical. Feb2018 Part 3, Vol. 255, p2589-2594. 6p.
Publication Year :
2018

Abstract

As one of the DNA damage repair enzymes, uracil-DNA glycosylase (UDG) plays an important role in maintaining genomic integrity. By linked bridge hybridizing-induced split G-quadruplex (SQ), we demonstrate here the construction of a simple and sensitive DNA machine for the detection of UDG activity. The satisfactory split G-quadruplex sequences (SQS) were successfully selected to be the ingredients of SQ formation. In this work, UDG recognized and removed the uracil bases from the stem of hairpin DNA (HP). And then, HP with a low melting temperature hybridized with designed SQS, forming a three-way DNA structure with an SQ. With the addition of Thioflavin T, a dramatical enhancement of fluorescence intensity was presented due to the G-quadruplex/Thioflavin T complex formation. The detection limit was as low as 7.8 × 10 −3 U/mL. And we also successfully investigated the performance of UDG activity in the HeLa cell lysate. This optical DNA machine with the merits of being simple, rapid, and economical was flexibly suitable for not only active UDG assay but also diverse target detection by adjusting the recognition region of the HP. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
09254005
Volume :
255
Database :
Academic Search Index
Journal :
Sensors & Actuators B: Chemical
Publication Type :
Academic Journal
Accession number :
126120166
Full Text :
https://doi.org/10.1016/j.snb.2017.09.065