A method for measuring 8-oxoguanine DNA glycosylase based on background signal suppression probe, kit and application thereof

A background signal and glycosylase technology, applied in the field of 8-oxoguanine DNA glycosylase determination, 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. problems, to achieve the effect of simple design, short detection time and low cost
CN113186248BActive Publication Date: 2022-04-29HUAZHONG UNIV OF SCI & TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
HUAZHONG UNIV OF SCI & TECH
Publication Date
2022-04-29

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Abstract

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. The present invention utilizes the selectivity of Lambda exonuclease to the substrate structure and the recognition and cleavage of 8-oxoguanine by 8-oxoguanine DNA glycosylase, combined with the background signal to suppress the effect of the probe on the side activity of λexo Inhibition, a new 8‑oxoguanine DNA glycosylase fluorescence assay method was constructed. Specifically include: mixing λexo buffer, reporter probe sequences containing fluorescent groups and quenching groups, and background signal suppression probe sequences containing 8-oxoguanine; heating the mixture to 85°C, and then cooling down gradually Add the sample to be tested at 37°C; incubate at 37°C; after the incubation, add λexo to carry out enzyme digestion reaction, and then perform 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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