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DTNS-mediated method for detecting activity of 8-OG DNA glycosylase

A DNA probe and tetrahedron technology, applied in the field of biological detection and molecular biology, can solve problems such as affecting detection accuracy and interference, and achieve the effect of preventing false positive signals

Active Publication Date: 2021-12-14
LIAOCHENG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Despite significant progress, these in situ fluorescence methods relying on the intensity of a single fluorescent signal are still subject to interference from false positive signals generated by intracellular nuclease degradation, affecting detection accuracy
Therefore, accurate and in situ imaging of intracellular 8-OG DNA glycosylase activity remains a challenge

Method used

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  • DTNS-mediated method for detecting activity of 8-OG DNA glycosylase
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  • DTNS-mediated method for detecting activity of 8-OG DNA glycosylase

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Embodiment Construction

[0025] It should be noted that the following detailed description is exemplary and intended to provide further explanation of the present invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0026] It should be noted that the terminology used here is only for describing specific embodiments, and is not intended to limit exemplary embodiments according to the present invention. As used herein, unless the context clearly dictates otherwise, the singular is intended to include the plural, and it should also be understood that when the terms "comprising" and / or "comprising" are used in this specification, they mean There are features, steps, operations, means, components and / or combinations thereof.

[0027] As introduced in the background technology, in the prior art, methods for detecting 8-OG DNA glycosylase activity usually only de...

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Abstract

The invention belongs to the technical field of biological detection and molecular biology, and particularly relates to a DTNS-mediated method for detecting the activity of 8-OGDNA glycosylase. Under the action of 8-OG DNA glycosylase, the structure of the DTNS is changed from an open state to a closed state, so that the distance between a Cy3 donor and a Cy5 receptor is close, and efficient FRET is initiated. By utilizing the method, the activity of extracellular 8-OG DNA glycosylase can be sensitively and selectively detected, and false positive signals generated by nuclease degradation can be prevented based on an FRET signal output mode, and accurate imaging in living cells is facilitated.

Description

technical field [0001] The invention belongs to the technical field of biological detection and molecular biology, and in particular relates to a method for detecting 8-OGDNA glycosylase activity mediated by DTNS. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] 8-Oxidative guanine (8-OG) DNA glycosylase is a DNA repair enzyme that repairs 8-OG, a common oxidative DNA damage. The enzyme recognizes 8-OG in double-stranded DNA and cleaves the glycosidic bond to generate an apurinic / apyrimidinic (AP) site, which is then excised by the enzyme's intrinsic AP-cleaving activity. However, the abnormal activity of 8-OG DNA glucoamylase in cells can lead to wrong base pairing betwe...

Claims

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Application Information

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IPC IPC(8): C12Q1/34C12Q1/6818C12Q1/6806C12N15/11C12N15/10G01N21/64G01N21/63
CPCC12Q1/34C12Q1/6818C12Q1/6806G01N21/6428G01N21/63G01N2333/924C12Q2521/531C12Q2565/1015C12Q2563/107C12Q2563/155Y02A50/30
Inventor 吴玉姝吴敏刘敏韩军
Owner LIAOCHENG UNIV
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