A new approach for the rapid detection of common and atypical aldehyde dehydrogenase alleles.
A strong protection against the development of alcoholism is exerted by a point mutation in the gene coding for low Km aldehyde dehydrogenase (ALDH), i.e. ALDH2. We report a non-radioactive method for determining the common and atypical human mitochondrial aldehyde dehydrogenase (ALDH2) genotypes. This method is based on the fact that the base change (G-->A) in Exon 12 of the ALDH2 gene abolishes an Eco57 I restriction site (CTGAAG-->CTAAAG). A GC-clamp attached oligonucleotide was designed to yield a 176 base pair product by the polymerase chain reaction. After amplification, the resulting fragment containing the normal nucleotide sequence is cut by Eco57 I into two segments (131 base pairs + 45 base pairs) while the fragment containing the mutated sequence remains intact (176 base pairs). These are visualized by staining with ethidium bromide on agarose gels without blotting, hybridization or autoradiography.
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Addiction Research Foundation of Ontario, Toronto, Canada.
Israel
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D000437:Alcoholism / Q000235:genetics; D000444:Aldehyde Dehydrogenase / Q000235:genetics*; D000483:Alleles; D001483:Base Sequence; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004996:Ethidium; D005091:Exons; D005838:Genotype; D006801:Humans; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D017354:Point Mutation; D016133:Polymerase Chain Reaction
D015345:Oligonucleotide Probes; D000444:Aldehyde Dehydrogenase; D004996:Ethidium