We use BSA115 strain which lacks RsbU and RsbW proteins. Therefore, there is limited post-transcriptional regulation of sigmaB activity.
SigmaB itself is placed downstream of Pspac, inducible by IPTG. The lacZ reporter gene is downstream of Pctc promoter.
IPTG concentrations of 0.1, 0.2 and 1 mM are added in mid-exponential phase at an OD of appr. 0.3. The whole experiment runs for about eight hours.
SEEK ID: https://fairdomhub.org/assays/21
Assay type: Proteomics
Technology type: Technology Type
Organisms: Bacillus subtilis (batch)
Created: 16th Nov 2009 at 16:10
Last updated: 8th Nov 2017 at 15:21
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Disciplines: Not specified
Roles: Not specified
Expertise: sensitivity analysis, Systems Biology, stress responses, Deterministic modelling of gene regulation networks, dynamics and control of biological networks, parameter estimation, Bacillus subtilis, Mathematical modelling
Modelling of the general stress response activation cascade of sigB in B. subtilis in response to starvation.
Modelling carbon core metabolism in Bacillus subtilis – Exploring the contribution of protein complexes in core carbon and nitrogen metabolism.
Bacillus subtilis is a prime model organism for systems biology approaches because it is one of the most advanced models for functional genomics. Furthermore, comprehensive information on cell and molecular biology, physiology and genetics is available and the European Bacillus community (BACELL) has a well-established reputation for applying
The objective of this project is an integrated understanding the metabolic, proteomic and genetic network that controls the transition from growth to glucose starvation. This transition is a fundamental ecophysiological response that serves as a scientific model for environmental signal integration and is pivotal for industrial fermentations of Bacillus that occur predominantly under nutrient starvation.
Glucose starvation, Transcriptomics, Proteomics, Metabolomics,Bacillus subtilis,
Snapshots: No snapshots
Studies: B. subtilis Transcription Factor Competition, Batchfermentation exp-starv01_090204, Biphase Batch Fermentation(2009/02/04), Controlled sigmaB induction in shake flask, Transition to starvation in shake flask
Assays: 2D-gelbased analysis of intracellular proteins, Absolute quantification of proteins by the AQUA-technology, B. subtilis Transcription Factor Competition - theoretical interpretation, B. subtilis Transcription Factor Competition - theoretical interpretation, Fermentation-BM5_SysMo, Gene expression(Transcriptome), IPTG induction of sigmaB in BSA115, IPTG induction of sigmaB in BSA115, Relative quantification of proteins by metabolic labeling, Stressosome activation dynamics, metabolome-LCMS
In this kind of studies sigmaB stress response is induced by the addition of artificial inducers of sigmaB. For example simgaB is downstream of a Pspac promoter and induced by the addition of IPTG.
A ctc::lacZ reporter gene is used to monitor sigmaB activity.
Person responsible: Ulf Liebal
Snapshots: No snapshots
Strain BSA115 is grown until appr. OD 0.25 then expression of sigB is induced by the addition of IPTG. The extend of stress response is measured by the expression of lacZ via beta-Gal assay. The experiment lasts for appr. 400 min.
SOP for ß-Galactosidase assay.
Investigations: The transition from growing to non-growing Baci...
SOP for shake flask cultivation of B.Subtilis in Bacell-Sysmo
Date Published: 2012
Journal: Mol. BioSyst.