Fluorescence chemical sensor for detecting DNA adenine transmethylase, and detection method thereof

Abstract

The invention discloses a fluorescence chemical sensor for detecting Dam transmethylase. The fluorescence chemical sensor for detecting the Dam transmethylase comprise a Dam transmethylase detection probe and an auxiliary probe; the Dam transmethylase detection probe is a hairpin DNA probe with stem-loop structures; the two stem structures are complemented to form double-chain DNA; the double-chain DNA has an identification site of the Dam transmethylase; the 3' terminal of the hairpin DNA probe is modified by amino; the identification site of the Dam transmethylase is a palindromic sequence of 5'-G-A-T-C-3'; and the auxiliary probe is a T-enriched single-chain DNA sequence with the near 3' terminal with a apurinic pyridine site (AP site). The preparation and detection method of the fluorescence chemical sensor is simple, the detection sensitivity is greatly improved, nonspecific reaction is effectively avoided, and the Dam transmethylase can be detected accurately in a complex biological sample.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, in particular to a fluorescent chemical sensor based on a hyperbranched amplification zero-background detection of DNA adenine methyltransferase and a detection method thereof. Background technique [0002] DNA methylation is the most prominent form of epigenetic gene regulation and plays a key role in cell proliferation, gene transcription and aging. Covalent addition of methyl groups to targeted adenine / cytosine residues in the recognition sequence, catalyzed by DNA methyltransferase, using S-adenosyl-L-methionine (SAM) as a methyl donor . DNA methylation usually occurs at the C-5 / N-4 position of cytosine and the N-6 position of adenine. Hypermethylation and hypomethylation have been found in various types of cancers such as breast, ovarian and lung cancers. The abnormality of DNA methylation is related to the abnormal expression and activity of DNA methyltransferase. Therefore,...

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Problems solved by technology

However, they generally require tedious nanoparticle preparation, rely on external labeling with fluorophores and quenchers, have long analysis times, involve complex designs, and are expensive

The electrochemical method has fast response and low design cost, but the complex electrode surface modification limits its application

The introduction of nucleic acid amplification greatly improves the sensitivity of detection. However, the existing nucleic acid amplification usually requires DNA templates, specific recognition sequences for endonucleases, and fluorescent labels for nucleic acid substrates, which increase the difficulty of protocol design. and the complexity of the experimental operation

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