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PMID: 6298188 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

chlC (nar) operon of Escherichia coli includes structural genes for alpha and beta subunits of nitrate reductase.

Journal of bacteriology ·Vol. 153 ·No. 3 ·1983-03-00 ·Pages 1513-20

Edwards ES, Rondeau SS, DeMoss JA

Abstract

The synthesis of the alpha and beta subunits of nitrate reductase by 20 chlC::Tn5 insertion mutants of Escherichia coli was determined by immune precipitation of the subunits from fractions of cell extracts. Only two of the mutants produced either subunit in detectable amounts; these two accumulated the alpha subunit, but no beta subunit. In both cases the alpha subunit was present in the cytosolic fraction, in contrast to wild-type cells, in which both subunits are present mainly in the membrane fraction. EcoRI restriction fragments containing the Tn5 inserts from five of the mutants were cloned into pBR322. The insertions were localized on two contiguous EcoRI fragments spanning a 5.6-kilobase region that overlapped the contiguous ends of the two fragments. An insertion that permitted alpha subunit formation defined one end of the 5.6-kilobase region. The results indicated that the genes encoding the alpha and beta subunits of nitrate reductase were part of a chlC (nar) operon that is transcribed in the direction alpha leads to beta.

MeSH Terms
DNA Transposable Elements DNA, Bacterial/analysis Electrophoresis, Agar Gel Escherichia coli/genetics Genes Macromolecular Substances Mutation Nitrate Reductases/genetics Nucleic Acid Hybridization Operon
Chemicals
DNA Transposable Elements DNA, Bacterial Macromolecular Substances Nitrate Reductases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Edwards E S
Rondeau S S
DeMoss J A
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33 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.153.3.1513-1520.1983
Published
1983-03-00
Pages
1513-20
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC221803
Subset
IM
Grants
NIGMS NIH HHS · GM 07542 · United States
NIGMS NIH HHS · GM 19511 · United States
External Links
PubMed source DOI: 10.1128/jb.153.3.1513-1520.1983 PMC Full Text View journal details
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