A method for specific regulation of nuclease activity

A technology with nuclease activity and specificity, applied in the field of DNA sequence detection, can solve the problems of normal operation of the interferase-nucleic acid probe system, unclear distribution mechanism of nuclease, and reduced detection efficiency, so that the detection efficiency can be kept stable and preserved. The main activity and the effect of improving the detection efficiency
CN111172252BActive Publication Date: 2022-03-08HUAZHONG UNIV OF SCI & TECH
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Patent Information

Authority / Receiving Office
CN Β· China
Patent Type
Patents(China)
Current Assignee / Owner
HUAZHONG UNIV OF SCI & TECH
Publication Date
2022-03-08

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Abstract

The invention belongs to the technical field of DNA sequence detection, and discloses a method for specifically regulating nuclease activity, designing corresponding Guiding chains for gene sequences at different mutation sites; synthesizing and preparing the required Guiding chains; Mix with the probe detection system so that the ratio of Guiding strand to substrate strand is 2:1; add Endo IV to the above mixed system to convert the substrate strand into a longer DNA double strand, and perform real-time fluorescence measurement. The regulation of endonuclease IV / APE-1 / lambda exonuclease activity involved in the present invention reduces the background signal due to nuclease side activity, and at the same time retains the main activity of nuclease cutting the target sequence to the greatest extent, thereby improving detection efficacy.
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Description

technical field

[0001] The invention belongs to the technical field of DNA sequence detection, and in particular relates to a method for specifically regulating nuclease activity. Background technique

[0002] Nucleases are currently widely used tools in biological, chemical and medical research. They are widely used in the development of biosensors, gene editing technology and new drugs. Different nucleases have different cleavage properties and corresponding substrates are also different. At the level of practical application, the current application of nuclease is based on its predictable working mechanism. For example, we usually want Endo IV to be able to stably and specifically cut DNA duplexes with gaps. However, nucleases are easily interfered by other nucleic acid strands in the system, and their cleavage properties are not stable. Especially for nucleases that target double-stranded DNA, they often cleave single-stranded nucleic acids non-specifically. This ph...

Claims

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