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PMID: 2986130 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

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

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 9 ·1985-05-00 ·Pages 2703-7

Ikuta T, Fujiyoshi T, Kurachi K, Yoshida A

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 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 [Young, R. A. & Davis, R. W. (1983) Proc. Natl. Acad. Sci. USA 80, 1194-1198]. 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 (alpha, beta, and gamma) controlled by the ADH1, ADH2, and ADH3 loci. Ten lambda gt11 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 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.

MeSH Terms
Alcohol Dehydrogenase Alcohol Oxidoreductases/genetics,immunology Amino Acid Sequence Antibodies Bacteriophage lambda/genetics Base Sequence Chromosome Mapping Cloning, Molecular DNA/genetics DNA Restriction Enzymes Humans Liver/metabolism
Chemicals
Antibodies DNA Alcohol Oxidoreductases Alcohol Dehydrogenase DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ikuta T
Fujiyoshi T
Kurachi K
Yoshida A
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31 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.82.9.2703
Published
1985-05-00
Pages
2703-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC397633
Subset
IM
Grants
NIAAA NIH HHS · AA05763 · United States
NHLBI NIH HHS · HL-29515 · United States
Databases
GENBANK
K01883, M11831, M11832, M11833, M11834, M11835, M11836, M11837, M11838, M11839, M24317
External Links
PubMed source DOI: 10.1073/pnas.82.9.2703 PMC Full Text View journal details
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