Xu Y L, Carr L G, Bosron W F, Li T K, Edenberg H J
Department of Biochemistry, Indiana University School of Medicine, Indianapolis 46223.
Genomics. 1988 Apr;2(3):209-14. doi: 10.1016/0888-7543(88)90004-3.
Humans are polymorphic at two of the alcohol dehydrogenase (ADH) loci important in ethanol metabolism, ADH2 and ADH3. Although the coding regions of these genes are 94% identical, they produce subunits that differ greatly in kinetic properties in vitro. These differences are likely to be reflected in the pharmacokinetics of alcohol metabolism, but studies have been hampered by the need to use liver biopsy specimens to determine the ADH phenotype. This problem has now been overcome by determining the genotype at these loci using DNA that has been amplified in vitro by the polymerase chain reaction. We report here the identification of all three of the ADH2 alleles and both of the ADH3 alleles. Any pair of ADH2 or ADH3 alleles can be distinguished using allele-specific oligonucleotide probes directed at their single base pair difference. In addition, ADH2(2) can be distinguished from ADH2(1) and ADH2(3) by detecting a new MaeIII site created in the third exon by the single base pair alteration in ADH2(2).
人类在乙醇代谢中起重要作用的两个乙醇脱氢酶(ADH)基因座,即ADH2和ADH3上存在多态性。尽管这些基因的编码区有94%的同一性,但它们产生的亚基在体外动力学特性上有很大差异。这些差异可能反映在酒精代谢的药代动力学中,但由于需要使用肝活检标本确定ADH表型,相关研究受到了阻碍。现在,通过使用经聚合酶链反应体外扩增的DNA来确定这些基因座的基因型,这个问题已得到解决。我们在此报告ADH2的所有三个等位基因以及ADH3的两个等位基因的鉴定情况。针对ADH2或ADH3等位基因之间单碱基对差异设计的等位基因特异性寡核苷酸探针,可区分任意一对ADH2或ADH3等位基因。此外,通过检测ADH2(2)中单个碱基对改变在第三个外显子中产生的一个新的MaeIII位点,可将ADH2(2)与ADH2(1)和ADH2(3)区分开来。