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Detection method for allele specific methylation

An allele-specific and detection method technology, applied in the field of allele-specific methylation detection, can solve the problems of gene transcription inactivation, uncontrolled cell proliferation of synthesized mature functional proteins, and hindering the combination of transcription factors and promoters, etc.

Inactive Publication Date: 2014-03-26
仲集华
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  • Application Information

AI Technical Summary

Problems solved by technology

Methylation of the promoter region of tumor suppressor genes can hinder the binding of transcription factors to the promoter, resulting in inactivation of gene transcription, blocking the synthesis of mature functional proteins, and leading to uncontrolled cell proliferation

Method used

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Embodiment Construction

[0009] Firstly, all CCGG sequences and SNP sites were found in the promoter region and the first exon region of TIMP3 gene, and then allele-specific PCR technology and methylation-sensitive restriction enzyme digestion-PCR method were combined to detect a specific expression of TIMP3. Specific detection of the methylation status of the CpG islands in the promoter regions of alleles. The SNP must be located in the transcription region; the SNP is in the promoter region; and the SNP is in or the first exon region.

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Abstract

The invention discloses a detection method for allele specific methylation. The method comprises the following steps: finding all CCGG sequences and SNP (Single Nucleotide Polymorphism) loci in a TIMP3 gene promoter region and a first exon region; performing specific detection on the methylation state of a pair of allele promoter region CpG islands of TIMP3 by combining an allele specific PCR (Polymerase Chain Reaction) technology and a methylation-sensitive restriction digestion-PCR method, wherein the SNP is located in a transcription region; the SNP is located in the promoter region; or the SNP is located in the first exon region. The method has the beneficial effects that the out-of-control situation of cell proliferation due to hindering of the synthesis of mature functional proteins under the situation of transcriptional inactivation caused by the combination of transcription factors and promoters can be hindered effectively.

Description

technical field [0001] The invention relates to a biochemical detection method, in particular to a detection method for allele-specific methylation. Background technique [0002] DNA methylation is one of the most important epigenetic modifications and is closely related to tumorigenesis. Methylation of the promoter region of tumor suppressor genes can hinder the combination of transcription factors and promoters, resulting in inactivation of gene transcription, blocking the synthesis of mature functional proteins and resulting in uncontrolled cell proliferation. The research on methylation inactivation of tumor suppressor gene has important guiding significance in the early diagnosis, treatment and prognosis of clinical tumors, and is a current research hotspot. We have previously detected the methylation of multiple sites in gastric cancer in the high-incidence area of ​​southern Liaoning, and found that the frequency of DNA methylation in the promoter regions of specific...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858
Inventor 仲集华
Owner 仲集华