PCR-RFLP method for detecting single-nucleotide polymorphism of type II diabetes susceptibility gene CREB1 and application thereof

Abstract

The invention discloses a PCR-RFLP method for detecting the single-nucleotide polymorphism of the type II diabetes susceptibility gene CREB1 and application thereof. According to the invention, the detected gene polymorphism comprises the single-nucleotide polymorphism of T or G located in the promoter region of the CREB1 gene and -1354 bp away from a transcription start site and the single-nucleotide polymorphism of T or A located in the promoter region of the CREB1 gene and -1343 bp away from the transcription start site according to the results of sequencing with DNA pools; the genome DNAs of a to-be-detected sample are used as a template, a primer pair P is used as a primers, and PCR amplification is carried out on a CREB1 gene sequence containing polymorphic sites in the presence of Taq DNA polymerase, Buffer (a buffer environment), Mg<++> and dNTPs; then a PCR product is divided into two parts which are respectively digested by using Hha I and Xsp I restriction enzymes; and enzyme-digested products are subjected to typing via agarose gel electrophoresis. Moreover, the above two polymorphic sites and clinical indicators of type II diabetes are subjected to association analysis, and results show that different genotypes are substantially related to the levels of fasting blood glucose and glycosylated hemoglobin.

Description

technical field [0001] The invention relates to the field of molecular biology detection of human diseases, in particular to a method for detecting type II diabetes susceptibility genes CREB1 Single nucleotide polymorphism PCR-RFLP method and its application. Background technique [0002] Diabetes is considered to be the fifth most serious disease that endangers human health in the world, and 90% of them are non-insulin-dependent diabetes, also known as type II diabetes. The disease is a complex metabolic disorder, and its pathogenesis is complex, which is the result of the interaction between genetics and the environment, among which genetic polymorphism may be the main factor affecting the susceptibility of the disease among individuals. Since the early clinical symptoms of type II diabetes are not obvious, patients often delay the best time for treatment. Therefore, finding and identifying genetic markers related to type 2 diabetes in the human genome will be beneficial...

AI Technical Summary

Problems solved by technology

[0007] Currently, domestic and foreign CREB1 The study of genes mainly focuses on their functions and regulatory mechanisms. CREB1 Genetic variants and their association with type 2 diabetes have not been reported

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