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PMID: 2376568 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Isolation and characterization of Rhodobacter capsulatus mutants defective in oxygen regulation of the puf operon.

Journal of bacteriology ·Vol. 172 ·No. 8 ·1990-08-00 ·Pages 4549-54

Narro ML, Adams CW, Cohen SN

Abstract

cis-acting mutations that affect regulation of the Rhodobacter capsulatus puf operon by oxygen were isolated by placing the mutagenized puf regulatory region 5' to a promoterless Tn5 neo gene, which encodes resistance to kanamycin (Kmr). R. capsulatus mutants that failed to show wild-type repression of KMr by oxygen were selected and analyzed. Four independent clones contained point mutations, three of which were identical, in a region of dyad symmetry located between puf operon nucleotide positions 177 and 207, approximately 45 base pairs 5' to the site of initiation of puf transcripts. The phenotypic effects of the aerobically selected mutations were duplicated by single and double point mutations introduced site specifically into the region of dyad symmetry by oligonucleotide-directed mutagenesis. Determinations of the bacterial 50% lethal dose of kanamycin, of aminoglycoside phosphotransferase activity in cell sonicates, and of neo-specific mRNA confirmed the diminished responsiveness of the mutants to oxygen and consequently implicated the mutated region in O2-mediated transcriptional regulation.

MeSH Terms
Base Sequence Cloning, Molecular Gene Expression Regulation, Bacterial/drug effects Molecular Sequence Data Mutation Oligonucleotide Probes Operon/drug effects Oxygen/pharmacology Plasmids Promoter Regions, Genetic Restriction Mapping Rhodopseudomonas/drug effects,genetics
Chemicals
Oligonucleotide Probes Oxygen
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Narro M L
Department of Genetics, Stanford University School of Medicine, California 94305.
Adams C W
Cohen S N
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.8.4549-4554.1990
Published
1990-08-00
Pages
4549-54
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC213287
Subset
IM
Grants
NIGMS NIH HHS · GM 11225 · United States
NIGMS NIH HHS · GM 27241 · United States
NIGMS NIH HHS · T32 GM07790 · United States
External Links
PubMed source DOI: 10.1128/jb.172.8.4549-4554.1990 PMC Full Text View journal details
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