Parameters affecting gene expression from the Pm promoter in gram-negative bacteria.
The Pm promoter inserted chromosomally or in broad-host-range replicons based on plasmid RSF1010 or RK2 are useful systems for both high- and low-level expression of cloned genes in several gram-negative bacterial species. The positive Pm regulator XylS is activated by certain substituted benzoic acid derivatives, and here we show that these effectors induce expression of Pm at similar relative ranking levels in both Escherichia coli and Pseudomonas aeruginosa However, the kinetics of expression was not the same in the two organisms. Different carbon sources and dissolved oxygen levels displayed limited effects on expression, but surprisingly the pH of the growth medium was found to be of major importance. By combining the effects of genetic and environmental parameters, expression from Pm could be varied over a ten-thousand- to a hundred-thousand-fold continuous range, and as an example of its applications we showed that Pm can be used to control the xanthan biosynthesis in Xanthomonas campestris.
Winther-Larsen
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UNIGEN Center for Molecular Biology and Department of Biotechnology, Norwegian University of Science and Technology, Trondheim.
Josefsen
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Brautaset
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Valla
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D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015534:Trans-Activators / Q000235:genetics*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D015534:Trans-Activators; C078720:XylS protein, Pseudomonas putida