title: A new approach for the rapid detection of common and atypical aldehyde dehydrogenase alleles.
abstract: 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.
authors.affiliation: Addiction Research Foundation of Ontario, Toronto, Canada.
authors.forename: G C
authors.initials: GC
authors.lastname: Tu
authors.forename: Y
authors.initials: Y
authors.lastname: Israel
mesh_terms: 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
chemical_list: D015345:Oligonucleotide Probes; D000444:Aldehyde Dehydrogenase; D004996:Ethidium