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

Tohru Ikuta, T. Fujiyoshi, K. Kurachi, A. Yoshida

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

We have cloned a full-length cDNA coding for human alcohol dehydrogenase (ADH; alcohol:NAD+ oxidoreductase, EC 1.1.1.1) from a human liver cDNA library constructed in phage λgt11. 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 second probe. These amino acid sequences are considered to be common in all three subunits (α, β, and λ) controlled by the ADH1, ADH2, and ADH3 loci. Ten λgt11 recombinants of 35 positive plaques obtained by antibody screening contained inserted cDNAs of 1.5-2.4 kilobase pairs and were found to exhibit 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 β1 subunit, a chain initiation codon, a chain termination codon, and additional 3' and 5' untranslated regions. A complete amino acid sequence of the human β1 subunit was deduced from the cDNA.

Original languageEnglish (US)
Pages (from-to)2703-2707
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume82
Issue number9
DOIs
StatePublished - Jan 1 1985
Externally publishedYes

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Alcohol Dehydrogenase
Molecular Cloning
Complementary DNA
Amino Acid Sequence
Initiator Codon
Antibodies
Terminator Codon
5' Untranslated Regions
3' Untranslated Regions
Gene Library
Oligonucleotides
Base Pairing
Bacteriophages
Libraries
Rabbits
Amino Acids
Liver

ASJC Scopus subject areas

  • General

Cite this

Molecular cloning of a full-length cDNA for human alcohol dehydrogenase. / Ikuta, Tohru; Fujiyoshi, T.; Kurachi, K.; Yoshida, A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 82, No. 9, 01.01.1985, p. 2703-2707.

Research output: Contribution to journalArticle

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