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Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene

Goldberg, David A. (1980) Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene. Proceedings of the National Academy of Sciences of the United States of America, 77 (10). pp. 5794-5798. ISSN 0027-8424. doi:10.1073/pnas.77.10.5794.

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The alcohol dehydrogenase (ADH; alcohol:NAD+ oxidoreductase, EC gene (Adh) of Drosophila melanogaster was isolated by utilizing a mutant strain in which the Adh locus is deleted. Adult RNA from wild-type flies was enriched in ADH sequences by gel electrophoresis and then used to prepare labeled cDNA for screening a bacteriophage library of genomic Drosophila DNA. Of the clones that hybridized in the initial screen, one clone was identified that hybridized with labeled cDNA prepared from a wild-type Drosophila strain but did not hybridize with cDNA prepared from an Adh deletion strain. This clone was shown to contain ADH structural gene sequences by three criteria: in situ hybridization, in vitro translation of mRNA selected by hybridization to the cloned DNA, and comparison of the ADH protein sequence with a nucleotide sequence derived from the cloned DNA. Comparison of the restriction site maps from clones of three different wild-type Drosophila strains revealed the presence of a 200-nucleotide sequence in one strain that was absent from the other two strains. The ADH mRNA sequences were located within the cloned DNA by hybridization mapping experiments. Two intervening sequences were identified within Adh by S1 nuclease mapping experiments.

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Additional Information:© 1980 by the National Academy of Sciences. Communicated by Norman Davidson, July 10, 1980. There are more people who helped in various aspects of this work than can be acknowledged in this space. Special thanks go to Brian Seed and Randy Smith for their trenchant criticism and humor, Mark Silver for his inimitable technical skills, Jennie Cortenbach for preparing media, and Tip Benyajati, Bill Sofer, Mike Ashburner, and Bob Karp for making Adh a pleasant system to work on. Tom Maniatis, in particular, helped in getting me started, getting me thinking, and getting me space and money (a National Institutes of Health Program Project Grant and a National Science Foundation Predoctoral Fellowship). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:molecular cloning; polymorphism; intervening sequence
Issue or Number:10
Record Number:CaltechAUTHORS:GOLpnas80
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:7681
Deposited By: Tony Diaz
Deposited On:22 Mar 2007
Last Modified:08 Nov 2021 20:44

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