Alcohol Dehydrogenase in Drosophila: Isolation and Characterization of Messenger RNA and CDNA Clone

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1980 Dec 11

PMID 6780981

Citations 20

Authors

C BENYAJATI

N Wang

A Reddy

E Weinberg

W Sofer

Affiliations

Soon will be listed here.

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.

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A highly conserved sequence in the 3'-untranslated region of the drosophila Adh gene plays a functional role in Adh expression.

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