8-oxoguanine DNA glycosylase determination method based on background signal inhibition probe, kit and application thereof

A background signal, glycosylase technology, applied in the field of 8-oxoguanine DNA glycosylase assay, can solve the problem that the sensitivity of the detection system is difficult to meet the detection requirements, the preparation process of quantum dots is complicated, low cost and high sensitivity, etc. problem, to achieve the effect of simple design, short detection time and low cost
CN113186248AActive Publication Date: 2021-07-30HUAZHONG UNIV OF SCI & TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
HUAZHONG UNIV OF SCI & TECH
Publication Date
2021-07-30

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Abstract

The invention belongs to the technical field of biology, and particularly relates to an 8-oxoguanine DNA glycosylase determination method based on a background signal inhibition probe, a kit and an application thereof. According to the present invention, the selectivity of Lambda exonuclease on the substrate structure and the recognition cutting effect of 8-oxo guanine DNA glycosylase on 8-oxo guanine are utilized, and the inhibition effect of the background signal inhibition probe on the lambda exo side activity is combined so as to construct the new 8-oxo guanine DNA glycosylase fluorescence determination method. The method specifically comprises the following steps: mixing a lambda exo buffer solution, a reporter probe sequence containing a fluorophore and a quenching group, and a background signal inhibition probe sequence containing 8-oxo guanine; heating the mixed solution to 85 DEG C, then gradually cooling to 37 DEG C, and adding a sample to be detected; incubating at 37 DEG C; and after incubation is finished, adding lambda exo, carrying out enzyme digestion reaction, and then carrying out fluorescence detection.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a method for measuring 8-oxoguanine DNA glycosylase based on a background signal suppression probe, a kit and an application thereof. Background technique

[0002] 8-Oxoguanine is a common DNA oxidative damage caused by reactive oxygen species, which can easily cause gene mutation and seriously affect the health of living organisms. 8-oxoguanine DNA glycosylase (8-oxoguanine DNAglycosylase, OGG) is a key enzyme in the process of DNA repair. 8-oxoguanine DNA glycosylase has both N-terminal glycosylase activity and AP (apurinic / apyrimidinic)-lyase activity. N-terminal glycosylase activity can cleave damaged purine bases on double-stranded DNA, creating an AP site. AP-lyase activity cleaves either the 3' or 5' end of the AP site, creating a base gap with 3' and 5' phosphates. 8-oxoguanine DNA glycosylase can recognize and excise 8-oxoguanine, which is of great significanc...

Claims

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