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Vol. 180, Issue 13, 3421-3431, July 1, 1998
Department of Microbiology, The Technical
University of Denmark, DK-2800 Lyngby, Denmark
A gene homologous to the rpoS gene of Escherichia
coli was cloned from a Pseudomonas putida KT2440 gene
bank by complementation of the rpoS-deficient strain
E. coli ZK918. The rpoS gene of P. putida complemented the acid sensitivity and catalase deficiency of the rpoS mutant of E. coli and stimulated
expression of the RpoS-controlled promoter,
bolAp1. The gene was sequenced and found to be
highly similar to the rpoS genes of other gram-negative bacteria. Like in other gram-negative bacteria, a homolog of the nlpD gene was found upstream to the rpoS gene.
A transcriptional fusion of the promoter of the P. putida
rpoS gene to the luxAB genes from Vibrio
harveyi was constructed and used as an inactivated allele of
rpoS for gene replacement of the wild-type copy in the chromosome of P. putida. The resultant
rpoS mutant of P. putida, C1R1, showed
reduced survival of carbon starvation and reduced cross-protection
against other types of stress in cells starved for carbon, in
particular after a challenge with ethanol. Survival in soil
amended with m-methylbenzoate was also reduced in the mutant strain P. putida C1R1. The RpoS protein of
P. putida controls the expression of more than 50 peptides, which are normally expressed in cells after a short period of
carbon starvation.
Cloning, Sequencing, and Phenotypic
Characterization of the rpoS Gene from Pseudomonas
putida KT2440
Copyright © 1998 by American Society for Microbiology
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