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PMID: 6296769 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Deletions at intervening sequence splice sites in the alcohol dehydrogenase gene of Drosophila.

Nucleic acids research ·Vol. 10 ·No. 22 ·1982-11-25 ·Pages 7261-72

Benyajati C, Place AR, Wang N, Pentz E, Sofer W

Abstract

Two formaldehyde-induced, homozygous viable ADH-negative mutants, Adhfn4 and Adhfn6, possess no material that cross-reacts with antibody directed against ADH, no mature mRNA of wild-type size, and greatly reduced amounts of RNA that hybridizes with an Adh probe. We have cloned the genomic DNA sequences from these mutants in bacteriophage lambda Charon 4 and subcloned the Adh region into plasmid vector pBR327. Restriction analyses revealed one small deletion in each of these mutants and DNA sequencing showed that the splice junctions of the 65-base pair (bp) intervening sequence (IVS) were altered. Both cloned mutant Adh genes, as well as the wild-type gene, are capable of promoting correct specific transcription initiation in HeLa cell nuclear extracts in vitro. We conclude that Adhfn4 and Adhfn6 are defective in RNA processing. Our results provide evidence for the importance of the splice junction sequences in normal ADH RNA processing and stabilization in Drosophila. We also speculate that splicing of ADH RNA proceeds in a nonrandom manner: mutations in one of the intervening sequences appear to cause accumulation of a large ADH RNA containing at least one other IVS.

MeSH Terms
Alcohol Dehydrogenase Alcohol Oxidoreductases/genetics Animals Base Sequence Chromosome Deletion Cloning, Molecular DNA/metabolism DNA Restriction Enzymes Drosophila melanogaster/enzymology,genetics Formaldehyde/pharmacology Genes Mutation Plasmids Transcription, Genetic
Chemicals
Formaldehyde DNA Alcohol Oxidoreductases Alcohol Dehydrogenase DNA Restriction Enzymes
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Benyajati C
Place A R
Wang N
Pentz E
Sofer W
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37 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/10.22.7261
Published
1982-11-25
Pages
7261-72
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327002
Subset
IM
Grants
NIEHS NIH HHS · ES 02920 · United States
NIGMS NIH HHS · GM 28791 · United States
NCI NIH HHS · N01-CO-75380 · United States
Databases
GENBANK
J01066, M11290
External Links
PubMed source DOI: 10.1093/nar/10.22.7261 PMC Full Text View journal details
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