Document Detail


Isolation and characterization of Rhodobacter capsulatus mutants defective in oxygen regulation of the puf operon.
MedLine Citation:
PMID:  2376568     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
M L Narro; C W Adams; S N Cohen
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of bacteriology     Volume:  172     ISSN:  0021-9193     ISO Abbreviation:  J. Bacteriol.     Publication Date:  1990 Aug 
Date Detail:
Created Date:  1990-08-31     Completed Date:  1990-08-31     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2985120R     Medline TA:  J Bacteriol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  4549-54     Citation Subset:  IM    
Affiliation:
Department of Genetics, Stanford University School of Medicine, California 94305.
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MeSH Terms
Descriptor/Qualifier:
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*
Grant Support
ID/Acronym/Agency:
GM 11225/GM/NIGMS NIH HHS; GM 27241/GM/NIGMS NIH HHS; T32 GM07790/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Oligonucleotide Probes; 7782-44-7/Oxygen
Comments/Corrections

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