Specific primer and liquid phase chip for detecting MAP3K1 gene mutation

Abstract

The invention discloses a liquid phase chip and a specific primer for detecting MAP3K1 gene mutation. The liquid phase chip mainly comprises: ASPE primers, microspheres coated by different anti-tag sequences, and an amplification primer, wherein each ASPE primer is composed of a tag sequence at a 5' end and specific primer sequences at a 3' end and aiming at target gene mutation sites, and the specific primer sequences are as follows: SEQ ID NO.7 and SEQ ID NO.8 aiming at a T148G site, SEQ ID NO.9 and SEQ ID NO.10 aiming at an C87A site, and / or SEQ ID NO.11 and SEQ ID NO.12 aiming at a G260A site. The coincidence rate of the detection result of the detection liquid phase chip disclosed by the invention with that of a sequencing method reaches as high as 100%, and parallel detection of a wild type and a mutant of a plurality of mutation sites is achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for MAP3K1 gene mutation detection and a liquid phase chip. Background technique [0002] Mitogen-activated protein kinase kinase kinase 1 (mitogen activated protein kinase kinase kinase 1, MAP3K1), also known as MAPKKK1, MEKK or MEKK1, is located on the long arm of chromosome 5 5q11.2, specifically located at base pairs 56110899 to 56191978 on chromosome 5 between. The protein encoded by the MAP3K1 gene is a serine / threonine kinase that is part of several signal transduction cascades that include, in addition to the NF-κB signaling pathway, extracellular regulated protein kinase (ERK) and amino-terminal kinase ( JNK) pathway. The encoded protein is activated by autophosphorylation and requires the cofactor magnesium to phosphorylate other proteins. The study found that the MAP3K1 gene plays a key role in sex development...

AI Technical Summary

Problems solved by technology

[0003] At present, the methods for MAP3K1 gene mutation detection mainly include: Illumina fiber optic bead chip technology, Affymetrix SNP6. detection system, but the degree of automation is low, and there are many manual operations, which are difficult to meet the needs of practical applications.

Although the high-throughput makes the Affymetrix SNP6.0 chip technology relatively mature, it is not suitable for low- and medium-density clinical diagnostic chips, and it is difficult to expand the detection of SNPs or tags related to many biological traits in the same reaction system SNP, in addition, the Affymetrix SNP6.0 chip is mainly strong on the expression profile chip, with many species, relatively weak on the SNP chip, and the detection price is expensive, which cannot meet the actual needs

Fluorescent quantitative PCR technology has the characteristics of high sensitivity, strong specificity, and high degree of automation, but it also has the disadvantages of easy sample contamination and high false positive rate, and can only detect one mutation type at a time. The PCR-RFLP method is based on gene Changes in the recognition sites of restriction endonucleases caused by mutations, such as loss of sites or generation of new sites, amplify a specific fragment by PCR, then digest the amplified product with restriction endonuclease, and observe the fragments by electrophoresis Size, this method is used to detect gene mutations with altered restriction sites, and can directly determine the genotype, but this method cannot be used for the detection of gene mutations without new restriction sites

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