Fluorescent biosensor for detecting DNA methylated transferase as well as preparation and application thereof

Abstract

The invention discloses a fluorescent biosensor for detecting DNA methylated transferase and preparation and application thereof. The fluorescent biosensor comprises a symmetric double-ring dumbbell and a 3D tetrahedral fluorescent scaffold, the symmetric double-ring dumbbell contains DNA methylation sites, methylation double strands can be formed under catalysis of DNA methylated transferase, enzyme digestion reaction is performed to form two single-chain dumbbell rings with consistent structures; the top ends of the 3D tetrahedral fluorescent scaffold contain the same hairpin and are modified with a fluorophore and a quenching group, the fluorophore and the quenching group are close to each other to cause fluorescence quenching, the sheared dumbbell ring can be combined with the hairpin to generate fluorescence, and DNA methylated transferase with different concentrations can generate fluorescence signals with different intensities through catalytic reaction. The DNA methylated transferase detection method established on the basis of the sensor is good in stability and specificity and high in sensitivity, can be used for screening MTase inhibitors, and has important significance on early detection and diagnosis of clinical cancers, drug research and the like.

Description

technical field [0001] The invention relates to the technical field of fluorescent biosensor and biomarker detection, in particular to a fluorescent biosensor for detecting DNA methyltransferase and its preparation and application. Background technique [0002] DNA methylation refers to DNA methyltransferase using S-adenosylmethionine (SAM) as a methyl donor to carry out the 5' end carbon atom of CpG dinucleotide cytosine in the genome and on the DNA nucleic acid sequence. Catalyzes the epigenetic modification of a methyl group by covalently bonding it. DNA methylation plays an important role in genetic regulation such as X chromosome inactivation, genome imprinting and gene expression changes. DNA methylation is a common base covalent modification method in eukaryotes, and it is also the main epigenetic form in mammals. DNA methylation plays an important role in important genetic regulation such as X chromosome inactivation, genome imprinting and gene DNA methyltransferas...

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Problems solved by technology

[0004] In view of the shortcomings of the prior art described above, the object of the present invention is to provide a fluorescent biosensor for detecting DNA methyltransferase and its preparation and application, which is used to solve the problem of detection of DNA methyltransferase in the prior art. Method stability and specificity are not good

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