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PMID: 3290902 Published · ppublish English 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.

The copper, zinc-superoxide dismutase gene of Saccharomyces cerevisiae: cloning, sequencing, and biological activity.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 13 ·1988-07-00 ·Pages 4789-93

Bermingham-McDonogh O, Gralla EB, Valentine JS

Abstract

The gene for copper, zinc-superoxide dismutase (Cu,Zn-SOD; EC 1.15.1.1) from the yeast Saccharomyces cerevisiae has been cloned, sequenced, and shown to have physiological activity. The gene was isolated from a lambda gt11 library by using a long, unique deoxyoligonucleotide probe. The probe sequence was deduced from the known amino acid sequence by using a computer-generated yeast codon preference table. The sequence of the coding and flanking regions is reported. The cloned gene was expressed and shown to be active in vivo. A 3.2-kilobase fragment containing the coding region and 160 upstream bases, subcloned in a yeast/Escherichia coli shuttle vector, was used to transform a yeast strain lacking Cu,Zn-SOD activity. The presence of the Cu,Zn-SOD gene-containing plasmid corrected the characteristic dioxygen sensitivity of this strain. Electrophoretic transfer blots with antibody to yeast Cu,Zn-SOD showed the presence of the protein in transformants and wild-type yeast but not in the mutant. The role of Cu,Zn-SOD in defense against dioxygen toxicity is discussed in the light of these findings.

MeSH Terms
Amino Acid Sequence Base Sequence Fungal Proteins/genetics,metabolism Molecular Sequence Data Recombinant Proteins/metabolism Saccharomyces cerevisiae/enzymology,genetics Superoxide Dismutase/genetics,metabolism
Chemicals
Fungal Proteins Recombinant Proteins Superoxide Dismutase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Bermingham-McDonogh O
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.
Gralla E B
Valentine J S
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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.85.13.4789
Published
1988-07-00
Pages
4789-93
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC280521
Subset
IM
Grants
NIGMS NIH HHS · GM28222 · United States
Databases
GENBANK
J03279
External Links
PubMed source DOI: 10.1073/pnas.85.13.4789 PMC Full Text View journal details
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