Molecular cloning of a full-length cDNA for human alcohol dehydrogenase.

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Abstract
A full-length c[complementary]DNA coding for human alcohol dehydrogenase (ADH; alcohol:NAD+ oxidoreductase, EC 1.1.1.1) was cloned from a human liver cDNA library constructed in page .lambda.gt11. The library was screened by using a rabbit antibody against human ADH as a first probe, by the modified method of Young and Davis. Mixed 14-mer synthetic oligonucleotides encoding Asp-Asp-His-Val-Val and Gln-Cys-Gly-Lys-Cys were used as a 2nd probe. These amino acid sequences are considered to be common in all 3 subunits (.alpha., .beta. and .gamma.) controlled by the ADH1, ADh2, and ADH3 loci. Ten .lambda.gt11 recombinants of 35 positive plaques obtained by antibody secreening contained inserted cDNA of 1.5-2.4 kilobase pairs and were found to exhibited positive signals by hybridization with synthetic probes. One of them, with an inserted cDNA of 1631 base pairs, contained a sequence that encodes 374 amino acid residues of the human .beta.1 subunit, a chain initiation, codon, a chain termination codon, and additional 3'' and 5'' untranslated regions. A complete amino acid sequence of the human .beta.1 subunit was deduced from the cDNA.