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人类摄入酒精主要通过Ⅰ型酒精脱氢酶和乙醛脱氢酶共同催化。为建立乙醛脱氢酶基因２（ＡＬＤＨ２）的基因型检测方法，本研究应用限制性片段长度多态性分析法（ＲＦＬＰ－ＰＣＲ）测定ＡＬＤＨ２的基因型。根据ＡＬＤＨ２的基因序列资料，设计一含一个碱基替代的限制性内切酶ＭｂｏⅡ的可识别位点。随后ＤＮＡ模板在下列条件扩增：变性；９４℃，１ｍｉｎ。退火；５７℃，３ｍｉｎ。延伸；７２℃，１ｍｉｎ。３５个循环。聚合酶链反应产物经１ＵＮＩＴ的ＭｂｏⅡ于３７℃消化过夜。消化产物经电泳分析后得到三种清晰的ＡＬＤＨ２基因型图谱。结果表明该法简单、快速、准确，有助于ＡＬＤＨ２基因型的检测
Ingestion of alcohol by humans is mainly catalyzed by type I alcohol dehydrogenase and aldehyde dehydrogenase. In order to establish a genotypic assay for aldehyde dehydrogenase gene 2 (ALDH2), the genotype of ALDH2 was determined by restriction fragment length polymorphism (RFLP-PCR). According to the ALDH2 gene sequence data, design a recognition site containing one base substitution MboII restriction endonuclease. The DNA template was then amplified under the following conditions: denaturation; 94 ° C, 1 min. Annealing; 57 ° C, 3 min. Extension; 72 ° C, 1 min. 35 cycles. Polymerase chain reaction products were digested overnight at 37 ° C with 1 UNIT Mbo II. Digestion products electrophoretic analysis of the three clear ALDH2 genotype map. The results show that the method is simple, rapid and accurate, which is helpful for the detection of ALDH2 genotype