Manganese dioxide thin sheet mimic enzyme sensor and its preparation method and detection method of t4pnk

Abstract

The invention discloses a manganese dioxide sheet mimic enzyme sensor and a preparation method thereof as well as a T4PNK detection method. The preparation method comprises the following steps: (1) mixing a MnO2 nanosheet solution with a TMB (3,3',5,5'-tetramethyl benzidine) solution to obtain a MnO2-TMB mixed solution; (2) dissolving hairpin DNA in a Tris-HCl buffer solution, and adding ATP (adenosine triphosphate) and lambda EXO (lambda exonuclease) for mixing to obtain a DNA solution; and (3) mixing the MnO2-TMB mixed solution and the DNA solution to obtain the manganese dioxide sheet mimic enzyme sensor. The preparation method is simple in operation, and the sensor has the characteristics of high sensitivity, low detection limit and simple operation when used for detecting T4PNK.

Description

technical field

[0001] The invention relates to a simulated enzyme sensor, in particular to a manganese dioxide thin slice simulated enzyme sensor, a preparation method and a T4PNK detection method. Background technique

[0002] T4 polynucleotide kinase (PNK), a 5' kinase, has been the most commonly used enzyme in molecular biology since it was first discovered in 1965 in E. coli protein extracts infected with even-numbered phages. It can catalyze the 5'-hydroxyl bearing of nucleotides or nucleotides at the 5' end of the phosphate residue of γ-adenosine triphosphate (ATP). This is very important for cellular nucleic acid metabolism, especially in the cellular response to DNA damage, which is closely related to many human diseases, such as Werner syndrome, Bloom syndrome and Rothmund-Thomson–Thomson syndrome. T4PNK is also widely used to detect DNA adducts or oligonucleotides and DNA damage repair. Therefore, the development of a sensitive and accurate assay for detecting T...

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