Determination of the impact of specific enzyme reactions and regulatory proteins using isogenic E. coli mutant strains
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Mutant strains which carry deletions of important metabolic enzymes, as well as mutant strains with altered regulation, need to adapt by changing fluxes or gene expression to compensate with the absence/differed concentration of these key enzymes. This may give new insights in the regulation of these pathways/enzymes.
SEEK ID: https://fairdomhub.org/studies/10
Steady state studies for different oxygen availability in Escherichia coli
Projects: SUMO
Study position:
Experimentalists: Sonja Steinsiek, Stefan Stagge
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Created: 17th Sep 2009 at 11:05
Last updated: 22nd Nov 2011 at 19:12
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Projects: SUMO
Institutions: Max Planck Institute for Dynamics of Complex Technical Systems
Expertise: Microbiology, Molecular Biology, E. coli, Biotechnology
Tools: qRT-PCR, Bioreactor experiments/Fermentation (batch, Chemostat), Enzyme assays, 2D-PAGE
SysMO is a European transnational funding and research initiative on "Systems Biology of Microorganisms".
The goal pursued by SysMO was to record and describe the dynamic molecular processes going on in unicellular microorganisms in a comprehensive way and to present these processes in the form of computerized mathematical models.
Systems biology will raise biomedical and biotechnological research to a new quality level and contribute markedly to progress in understanding. Pooling European research ...
Projects: BaCell-SysMO, COSMIC, SUMO, KOSMOBAC, SysMO-LAB, PSYSMO, SCaRAB, MOSES, TRANSLUCENT, STREAM, SulfoSys, SysMO DB, SysMO Funders, SilicoTryp, Noisy-Strep
Web page: http://sysmo.net/
"Systems Understanding of Microbial Oxygen responses" (SUMO) investigates how Escherichia coli senses oxygen, or the associated changes in oxidation/reduction balance, via the Fnr and ArcA proteins, how these systems interact with other regulatory systems, and how the redox response of an E. coli population is generated from the responses of single cells. There are five sub-projects to determine system properties and behaviour and three sub-projects to employ different and complementary modelling ...
Programme: SysMO
Public web page: http://www.sysmo.net/index.php?index=55
Organisms: Escherichia coli, Escherichia coli K-12
Changing the oxygen availability leads to an adaptation of Escherichia coli at different biological levels. After pertubation of oxygen in chemostat experiments the microorganism(s) will come back to another steady state. This investigation deals with these stationary responses of Escherichia coli within the aerobiosis scale. The change for different biological variables, in different areas of the organism like the electron transport chain, the TCA cycle or globally is investigated by wildtype ...
Submitter: Michael Ederer
Studies: Basic regulatory principles of Escherichia coli’s electron transport cha..., Determination of the impact of specific enzyme reactions and regulatory ..., Quantitative analysis of catabolic carbon and electron fluxes in E. coli..., The Escherichia coli steady state response to oxygen: from molecular int...
Assays: Analysis of by-product formation rates in MG1655, Analysis of gene expression rates at different aerobiosis levels via RT-PCR, ArcA phosphorylation at different aerobiosis levels (steady states), Characterization of E. coli MG1655 and ∆sdhC and ∆frdA isogenic mutant s..., Determination of intracellular metabolite concentrations, Determination of intracellular redox state by means of NAD/NADH ratio, Determination of intracellular redox state by means of ubiquinones (oxd/..., FNR activity at different aerobiosis levels (steady state), Kinetic modelling of Escherichia coli's electron transport chain, Kinetic modelling of Escherichia coli's electron transport chain coupled..., Literature Data from Alexeeva et al., J. Bacteriol., 2000, 2002, 2003, Measurement of cytochrome numbers, Physiological measurements from Sheffield chemostat, Steady State Oxygen Response of E. coli WT and two Electron Transport Ch..., Transcriptional profiling of steady states at different aerobiosis levels
Snapshots: No snapshots
This assay describes how to analyze gene expression rates via RT-PCR.
Submitter: Sonja Steinsiek
Assay type: Gene Expression Profiling
Technology type: qRT-PCR
Investigation: Steady state studies for different oxygen avail...
Organisms: Escherichia coli
SOPs: Evans Medium, SUMO chemostat conditions, SUMO mRNA isolation Epicentre
Data files: Gene expression rates at different aerobiosis l... and 3 hidden items
Snapshots: No snapshots
This document describes by-product formation rates measured in MG1655 at steady-state conditions in Infors-Multifors-Bioreactors.
Submitter: Sonja Steinsiek
Assay type: Metabolomics
Technology type: Technology Type
Investigation: Steady state studies for different oxygen avail...
Organisms: Escherichia coli
SOPs: DCW measurement, Evans Medium, Measurement of extracellular metabolites, SUMO chemostat conditions
Data files: 3 hidden items
Snapshots: No snapshots
The task of this assay is to determine the impact of oxygen availability on the concentrations of metabolites from different central metabolic pathways. The focus lies on metabolites connected to glycolysis, tri-carbon-acid-cycle and energy metabolism. All strains have been cultured and analysed according to the SOPs listed below
Submitter: Stefan Stagge
Assay type: Metabolomics
Technology type: Liquid Chromatography Mass Spectrometry
Investigation: Steady state studies for different oxygen avail...
Organisms: Escherichia coli
SOPs: DCW measurement, Quenching and Extraction of intracellular metab..., SUMO chemostat conditions
Data files: 3 hidden items
Snapshots: No snapshots
E. coli MG1655 and ∆sdhC and ∆frdA isogenic mutant strains were characterized in batch growth curves aerobic and anaerobically. Optical density, glucose consumption and by-product accumulation were measured during growth.
Submitter: Sonja Steinsiek
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Steady state studies for different oxygen avail...
Organisms: Escherichia coli : K-12/MG1655 (wild-type / wild-type)
SOPs: Evans Medium, Measurement of extracellular metabolites
Data files: 2 hidden items
Snapshots: No snapshots
This data file shows results from the different chemostat experiments.Gene expression rates are presented.
Creators: Katja Bettenbrock, Sonja Steinsiek
Submitter: Katja Bettenbrock
This method describes how one can quench metabolism of Escherichia coli and extract metabolites from many kinds of metabolite classes like: nucleotides, sugar-phosphates, organic acids ....
Creator: Stefan Stagge
Submitter: Stefan Stagge
Creator: Sonja Steinsiek
Submitter: The JERM Harvester
Investigations: Analysis of Escherichia coli with linear electr..., Analysis of the glucose transport in Escherichi..., Steady state studies for different oxygen avail...
Studies: Analysis of Escherichia coli strains with linea..., Characterization of mutant strains with defects..., Determination of the impact of specific enzyme ...
Assays: Aerobic and anaerobic characterization of MG165..., Analysis of by-product formation rates in MG1655, Characterization of E. coli MG1655 and ∆sdhC an..., Determination of by-product formation and gluco...
Creator: Katja Bettenbrock
Submitter: The JERM Harvester
Investigations: Analysis of Escherichia coli with linear electr..., Dynamical studies for different oxygen availabi..., Steady state studies for different oxygen avail...
Studies: Analysis of Escherichia coli strains with linea..., Determination of the impact of specific enzyme ..., The Escherichia coli dynamical response to oxyg..., The Escherichia coli steady state response to o...
Assays: Analysis of by-product formation rates in MG1655, Analysis of gene expression rates at different ..., ArcA phosphorylation at different aerobiosis le..., Characterization of E. coli MG1655 and ∆sdhC an..., FNR activity at different aerobiosis levels (st..., Gene expression analysis of mutants with linear..., Physiological measurements from Sheffield chemo..., Transcriptional profiling of E. coli during ana...
Creator: Martijn Bekker
Submitter: The JERM Harvester
Investigations: Steady state studies for different oxygen avail...
Studies: Determination of the impact of specific enzyme ..., The Escherichia coli steady state response to o...
Assays: Analysis of by-product formation rates in MG1655, Determination of intracellular metabolite conce..., Determination of intracellular redox state by m..., FNR activity at different aerobiosis levels (st..., Physiological measurements from Sheffield chemo...
This file describes how to isolate mRNA from E. coli using the kit from Epicentre, for gene expression analysis via RT-PCR
Creator: Sonja Steinsiek
Submitter: Sonja Steinsiek
Investigations: Analysis of Escherichia coli with linear electr..., Analysis of the glucose transport in Escherichi..., Steady state studies for different oxygen avail...
Studies: Analysis of Escherichia coli strains with linea..., Characterization of mutant strains with defects..., Determination of the impact of specific enzyme ...
Assays: Aerobic and anaerobic characterization of MG165..., Analysis of gene expression rates at different ..., Characterization of MG1655 and mutant strains u..., Gene expression analysis of mutants with linear...
This document describes the chemostat conditions to run comparable experiments in different bioreactors in different labs.
Creator: Sonja Steinsiek
Submitter: Sonja Steinsiek
Investigations: Analysis of Escherichia coli with linear electr..., Analysis of the glucose transport in Escherichi..., Dynamical studies for different oxygen availabi..., Steady state studies for different oxygen avail...
Studies: Analysis of Escherichia coli strains with linea..., Characterization of mutant strains with defects..., Determination of the impact of specific enzyme ..., The Escherichia coli dynamical response to oxyg..., The Escherichia coli steady state response to o...
Assays: Analysis of by-product formation rates in MG1655, Analysis of gene expression rates at different ..., ArcA phosphorylation at different aerobiosis le..., Characterization of MG1655 and mutant strains u..., Determination of by-product formation and gluco..., Determination of intracellular metabolite conce..., Determination of intracellular redox state by m..., Gene expression analysis of mutants with linear..., Physiological measurements from Sheffield chemo..., Transcriptional profiling of E. coli during ana...
Abstract (Expand)
Editor:
Date Published: 27th Mar 2014
Publication Type: Not specified
PubMed ID: 24723921
Citation:
Abstract
Date Published: 1st Jun 2011
Publication Type: Not specified
DOI: 10.1016/j.jbiotec.2011.03.015
Citation: