Rapid detection method for MEK1 gene mutation

Abstract

The invention provides a rapid detection method for mitogen activated protein kinase 1 (Mitogen Activated protein Kinase Kinase 1, MEK1 / MAPKK1) gene mutation. The rapid detection method is realized by the following steps: designing a pair of wild primers at the two ends of an MEK1 gene mutation site and designing a pair of mutation primers on the mutation site; respectively designing wild primers and mutation types of a No.2 exon (MEK1-E2) and a No.3 exon (MEK1-E3) of MEK1; respectively amplifying a wild type template and a mutation type template by the primers; utilizing the wild primers to carry out PCR (Polymerase Chain Reaction) amplification on the two templates; respectively amplifying corresponding wild target segment and a mutant type target segment; mixing the two segments according to different ratios and distinguishing by using an HRM method. Compared with the other methods, the rapid detection method has the advantages of high specificity, high sensitivity, convenience and portability; the flux is high and the cost of each time is low.

Description

Technical field: [0001] The invention relates to the fields of biotechnology and medicine, especially molecular biology, molecular diagnosis and real-time quantitative PCR technology. Background technique: [0002] The MEK1 gene is located on human chromosome 15q22.1-22.33. The full length of the gene is about 104kb, and the mRNA length after transcription is 2603bp. It encodes a mitogen-activated protein kinase kinase consisting of 393 amino acid residues, which is a kind of serine / threonine protein kinase. [0003] MEK1 protein participates in MAPK cascade signal transduction, it is activated by MAPKKK, and activated MEK1 activates MAPK through covalent modification, that is, two-site phosphorylation. After MAPK is activated, it phosphorylates downstream substrates (transcription factors, protein kinases, enzymes, structural proteins, etc.) to regulate cell growth, proliferation, differentiation, apoptosis, adhesion, migration and other processes, thereby affecting tumori...

AI Technical Summary

Problems solved by technology

[0015] First: the sensitivity is not high, and a low proportion of mutations (<20%) cannot be detected;

[0016] Second: it takes a long time, usually 2-3 days;

[0017] Third: high cost

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