PCR-RFLP法鉴定人ALDH2基因型

Identification of Human ALDH2 Genotype by PCR-RFLP Method

摘要: ALDH2编码乙醛脱氢酶，是人体乙醇代谢途径的重要组分。人类ALDH2基因存在SNP (单核苷酸多态性)现象，其gDNA在37,030位发生单碱基替换后导致乙醛脱氢酶失活，且该突变基因在东亚人群中有较高频率。根据此单碱基的差别，本文采用PCR-RFLP方法，通过提取、纯化、酶切受试者口腔上皮细胞基因组DNA，实现了对于三种ALDH2基因型的鉴定，其创新点在于简化实验步骤、提升产物回收效率的同时使判断更加准确。结论：PCR-RFLP法鉴定人ALDH2基因型可作为分子遗传学实验教学的案例。

Abstract: ALDH2 encodes aldehyde dehydrogenase, which is an essential component of human ethanol metabolism. The SNP (Single Nucleotide Polymorphism) of human ALDH2 gene inactivates aldehyde dehydrogenase due to the replacement of base 37,030 in the genomic sequence, and the frequency of the mutant ALDH2 gene is particularly high in East Asian population. Aimed at the difference of this single base, this paper applies PCR-RFLP to distinguish the three ALDH2 genotypes after extraction, purification and enzymatic digestion of the genomic DNA from human oral epithelium. The innovation of this work is mainly reflected in simplifying the experiment procedures as well as improving the efficiency of DNA purification and the accuracy of genotyping. We propose that human ALDH2 genotyping by PCR-RFLP can be used as a typical sample of experimental teaching of molecular genetics.