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PMID: 6780981 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Alcohol dehydrogenase in Drosophila: isolation and characterization of messenger RNA and cDNA clone.

Nucleic acids research ·Vol. 8 ·No. 23 ·1980-12-11 ·Pages 5649-67

Benyajati C, Wang N, Reddy A, Weinberg E, Sofer W

Abstract

The mRNA for alcohol dehydrogenase (ADH) in D. melanogaster has been identified by translation in a cell-free system. The in vitro synthesized polypeptide, specifically precipitated by anti-ADH antibody, has identical subunit molecular weight (25,000 daltons) and tryptic peptide profile to the in vivo synthesized ADH. The poly A containing ADH-mRNA has been purified by specific precipitation of ADH-polysomes using anti-ADH antibody and S. aureus. Transformation of E. coli with the dA-tailed ADH-mRNA-complementary DNA hybrid annealed to the dT-tailed pBR322 yielded one plasmid which has been identified as the ADH-cDNA clone. The identification involved hybridization selection of ADH-mRNA and in vitro translation, in situ hybridization to the Adh locus on salivary gland polytene chromosomes and DNA sequencing. This ADH-cDNA plasmid contains 349 bases of the C-terminal protein coding and 180 bases of the 3' untranslated region.

MeSH Terms
Alcohol Oxidoreductases/genetics Animals Base Sequence Cloning, Molecular DNA/genetics Drosophila melanogaster/enzymology Mutation Nucleic Acid Hybridization Poly A RNA, Messenger/genetics
Chemicals
RNA, Messenger Poly A DNA Alcohol Oxidoreductases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Benyajati C
Wang N
Reddy A
Weinberg E
Sofer W
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35 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/8.23.5649
Published
1980-12-11
Pages
5649-67
Language
English
Region
England
NLM ID
0411011
PMCID
PMC324332
Subset
IM
Grants
NIEHS NIH HHS · ES-01527 · United States
NIGMS NIH HHS · GM-18257 · United States
NIGMS NIH HHS · GM21322-01 · United States
Databases
GENBANK
J01066, M11290
External Links
PubMed source DOI: 10.1093/nar/8.23.5649 PMC Full Text View journal details
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