Specific primers and liquid phase chip for detecting polymorphism of cyckin-dependent kinase 5 regulatorysubunit-associated protein 1-like 1(CDKAL1) gene

Abstract

The invention discloses specific primers and a liquid phase chip for detecting the polymorphism of a CDKAL1 gene. The liquid phase chip mainly comprises specific primers consisting of a tag sequence at a 5' end and specific primer sequences of polymorphic sites of a target gene, microspheres and amplification primers, wherein the specific primer sequences are one or more pairs from SEQ ID No.9 and SEQ ID No.10 for A107C, SEQ ID No.11 and SEQ ID No.12 for G323C, SEQ ID No.13 and SEQ ID No.14 for A210G and SEQ ID No.15 and SEQ ID No.16 for A75G; the tag sequence may be a sequence from SEQ ID No.1 to SEQ ID No.8. The prepared liquid phase chip for detecting the polymorphism of the CDKAL1 gene has a very high signal-to-noise ratio, and the cross reaction of a designed probe and an anti-tae sequence is prevented basically.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a CDKAL1 gene polymorphism detection specific primer and a liquid phase chip. Background technique [0002] CDK5 regulatory subunit-associated protein 1 analog 1 (cyclin-dependent kinase 5 regulatory subunit-associated protein 1-like 1, CDKAL1) gene is the coding gene of CDK5 (cyclin-dependent kinase 5) regulatory subunit-associated protein 1, located in the Region 22.3 of the short arm of chromosome 6, whose signal region is located on the fifth intron, is highly expressed in human pancreas and skeletal muscle cells. CDK5 plays an important role in protecting β-cell function and lowering blood sugar. CDK5 can cause β-cell degeneration under the action of high glucose toxicity, and the protein encoded by CDKAL1 gene can inhibit the activation of CDK5. If the CDKAL1 gene locus is mutated, the inhibitory effect on CDK5 will be weakened, resul...

AI Technical Summary

Problems solved by technology

Fluorescent quantitative PCR has the advantages of simple operation, quick results, and quantification. However, this technology has the disadvantages of easy sample contamination, cross-reaction, and high false positive rate; while PCR-RFLP method is based on restriction endonucleation caused by gene mutations. Changes in the enzyme recognition site, such as site loss or new site generation, a specific fragment is amplified by PCR, and then the amplified product is digested with a restriction endonuclease, and the size of the fragment is observed by electrophoresis. This method is used for Detection of gene mutations with altered restriction sites can directly determine the genotype, but this method cannot be used for the detection of gene mutations without new restriction sites

Again, the above methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

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