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T Schweder

Publications and source records attributed to T Schweder.

24 records · Page 2Linked to original sources

Regulation of Escherichia coli starvation sigma factor (sigma s) by ClpXP protease.

In Escherichia coli, starvation (stationary-phase)-mediated differentiation involves 50 or more genes and is triggered by an increase in cellular sigma s levels. Western immunoblot analysis showed that in mutants lacking the protease ClpP or its cognate ATPase-containing subunit ClpX, sigma s levels of exponential-phase cells increased to those of stationary-phase wild-type cells. Lack of other potential partners of ClpP, i.e., ClpA or ClpB, or of Lon protease had no effect. In ClpXP-proficient cells, the stability of sigma s increased markedly in stationary-phase compared with exponential-phase cells, but in ClpP-deficient cells, sigma s became virtually completely stable in both phases. There was no decrease in ClpXP levels in stationary-phase wild-type cells. Thus, sigma s probably becomes more resistant to this protease in stationary phase. The reported sigma s-stabilizing effect of the hns mutation also was not due to decreased protease levels. Studies with translational fusions containing different lengths of sigma s coding region suggest that amino acid residues 173 to 188 of this sigma factor may directly or indirectly serve as at least part of the target for ClpXP protease.

ATP-Dependent Proteases↗

Escherichia coli K12 relA strains as safe hosts for expression of recombinant DNA.

Most Escherichia coli K12 strains survive for a relatively long time outside the laboratory. Under the same conditions the isoallelic E. coli K12 relA mutants die faster because they lack the stringent response. The killing rate is increased by using a plasmid-encoded suicide system consisting of the phage T7 lysozyme gene driven by the E. coli alkaline phosphatase gene promoter (phoA). Cells containing this system were rapidly and effectively killed as soon as phosphate was made limiting. The combination of the chromosomal relA mutation and a conditional suicide system of this type provides an effective means of biological containment for recombinant E. coli strains.

Alkaline Phosphatase↗

An expression vector system providing plasmid stability and conditional suicide of plasmid-containing cells.

A cloning vector system was constructed on the basis of the pBR322 derivative pEG1 by introducing the whole parB locus of plasmid R1 cloned behind the promoter of the alkaline phosphatase gene (phoA) of Escherichia coli. The parB locus in combination with the phoA promoter ensures both (i) plasmid stabilization due to the post-segregational killing of plasmid-free cells during growth and (ii) killing of the cells induced by the potential environmental signal phosphate limitation. This vector, therefore, appears to be a model system for increasing the stability of recombinant plasmids and for decreasing the potential risks in the application of recombinant bacteria in industrial fermentations.

Alkaline Phosphatase↗

Clustering of adenomas in the large intestine.

In a necropsy study of 68 men and 36 women with single or multiple adenomas of the large intestine the distribution of all adenomas within the bowel in individual patients was examined. Each adenoma was recorded both by its distance in centimetres from the caecal pole and by the intestinal segment in which it was found. Adenomas in individual patients were shown to occur significantly closer together than expected from the general distribution of adenomas within each age group. The statistically significant, but not very strong effect of clustering may be caused by the existence of one or more fields of the large intestine specific to individuals who have a higher than average propensity for neoplastic development.

Adenoma↗

Analysis of the expression and function of the sigmaB-dependent general stress regulon of Bacillus subtilis during slow growth.

Glucose-limited continuous cultures were used to analyze sigmaB activity at decreasing growth rates. Expression of the sigmaB-dependent genes gsiB and ctc started to increase at a growth rate of 0.2 h-1, and both genes were induced approximately fivefold at a growth rate of 0.1 h-1 as compared to expression at the maximal growth rate. However, maximal sigmaB activity was only reached when the growth stopped as a result of the exhaustion of the carbon and energy source glucose. During glucose-limited growth, increased expression of the general stress regulon at growth rates below 0.2 h-1 did not provide wild-type cells with a growth advantage over sigB mutants. Instead, expression of the stress regulon seems to constitute a significant burden during glucose-limited growth, resulting in a selective growth advantage of the sigB mutant as compared to the wild-type at a growth rate of 0.08 h-1.

Bacillus subtilis↗