The arcABC operon required for fermentative growth of Pseudomonas aeruginosa on arginine: Tn5-751-assisted cloning and localization of structural genes.

Details

Serval ID
serval:BIB_3538057168DC
Type
Article: article from journal or magazin.
Collection
Publications
Title
The arcABC operon required for fermentative growth of Pseudomonas aeruginosa on arginine: Tn5-751-assisted cloning and localization of structural genes.
Journal
Journal of General Microbiology
Author(s)
Lüthi E., Mercenier A., Haas D.
ISSN
0022-1287 (Print)
ISSN-L
0022-1287
Publication state
Published
Issued date
1986
Volume
132
Number
10
Pages
2667-2675
Language
english
Abstract
Pseudomonas aeruginosa is able to utilize L-arginine as the energy source for growth under anaerobic, nitrate-free conditions. Mutations in the chromosomal arcABC gene cluster specifying the inducible arginine deiminase pathway enzymes abolish fermentative growth on arginine. From two different arc::Tn5-751 insertion mutants of P. aeruginosa recombinant plasmids have been derived which carry a resistance marker of transposon Tn5-751 plus flanking parts of the arc region. These recombinant plasmids served to reconstruct in vitro the functional arcABC cluster on a 5.6 kb fragment, which was inserted into the broad-host-range vector pKT240. In P. aeruginosa this 5.6 kb segment complemented arcABC mutations in trans and contained the control region necessary in cis for arc enzyme induction by oxygen limitation and arginine. The results of subcloning experiments and transcriptional lacZ fusions, the polarity of transposon insertions and the effect of external promoters led to the conclusion that the structural genes arcA (for arginine deiminase), arcB (for catabolic ornithine carbamoyltransferase) and arcC (for carbamate kinase) are contiguous and transcribed in the same direction. Thus, the arcABC cluster appears to have the characteristics of an operon. In Escherichia coli the cloned arcABC genes were expressed at low, non-inducible levels; strong vector promoters enhanced arc expression up to 100-fold. This indicates that transcriptional initiation at the arc promoter(s) is poor in E. coli.
Keywords
Arginine/metabolism, Cloning, Molecular, DNA Transposable Elements, Genes, Genes, Bacterial, Hydrolases/metabolism, Operon, Ornithine Carbamoyltransferase/metabolism, Phosphotransferases/metabolism, Phosphotransferases (Carboxyl Group Acceptor), Pseudomonas aeruginosa/enzymology, Pseudomonas aeruginosa/genetics
Pubmed
Web of science
Create date
25/01/2008 17:01
Last modification date
20/08/2019 13:22
Usage data