Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Role of chaperones and a cytoplasmatic protease in recombinant protein production in E. coli at suboptimal growth temperature

ABSTRACT: Recent studies have revealed that at lower cultivation temperatures (25°C) much higher percentage of correctly folded recombinant proteins can be extracted from inclusion bodies. The goal of our research was to investigate mechanisms determining characteristics of non-classical inclusion bodies production using gene expression profiling. Two strains of recombinant E. coli [BL21 (DE3)] grown at three different temperatures (25°C, 37°C and 42°C) were included in experiment.

ORGANISM(S): Escherichia coli

PROVIDER: GSE25561 | GEO | 2010/11/24

SECONDARY ACCESSION(S): PRJNA134285

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Similar Datasets

Role of chaperones and a cytoplasmatic protease in recombinant protein production in E. coli at suboptimal growth temperature

Project description:Recent studies have revealed that at lower cultivation temperatures (25°C) much higher percentage of correctly folded recombinant proteins can be extracted from inclusion bodies. The goal of our research was to investigate mechanisms determining characteristics of non-classical inclusion bodies production using gene expression profiling. Two strains of recombinant E. coli [BL21 (DE3)] grown at three different temperatures (25°C, 37°C and 42°C) were included in experiment. Gene expression was studied in two recombinant strains (production and control strain), grown at three different temperatures (25, 37 and 42 oC) in three biological replicates. Cells were harvesed at OD=4, except for the cells grown at 25 oC, here they were harvested at OD=4 and OD=10.

2010-11-24 | E-GEOD-25561 | biostudies-arrayexpress

Metabolomic analysis of riboswitch containing E. coli recombinant expression system

Project description:In this study we have employed metabolomics approaches to understand the metabolic effects of producing enhanced green fluorescent protein (eGFP) as a recombinant protein in Escherichia coli cells. This metabolic burden analysis was performed against a number of recombinant expression systems and control strains and included: (i) standard transcriptional recombinant expression control system BL21(DE3) with the expression plasmid pET-eGFP, (ii) the recently developed dual transcriptional–translational recombinant expression control strain BL21(IL3), with pET-eGFP, (iii) BL21(DE3) with an empty expression plasmid pET, (iv) BL21(IL3) with an empty expression plasmid, and (v) BL21(DE3) without an expression plasmid; all strains were cultured under various induction conditions. The growth profiles of all strains together with the results gathered by the analysis of the Fourier transform infrared (FT-IR) spectroscopy data, identified IPTG-dependent induction as the dominant factor hampering cellular growth and metabolism, which was in general agreement with the findings of GC-MS analysis of cell extracts and media samples. In addition, the exposure of host cells to the synthetic inducer ligand, pyrimido[4,5-d] pyrimidine-2,4-diamine (PPDA), of the orthogonal riboswitch containing expression system (BL21(IL3)) did not display any detrimental effects, and its detected levels in all the samples were at similar levels, emphasising the inability of the cells to metabolise PPDA. The overall results obtained in this study suggested that although the BL21(DE3)-EGFP and BL21(IL3)-EGFP strains produced comparable levels of recombinant eGFP, the presence of the orthogonal riboswitch seemed to be moderating the metabolic burden of eGFP production in the cells enabling higher biomass yield, whilst providing a greater level of control over protein expression.

2017-11-16 | MTBLS393 | MetaboLights

Time series of Escherichia coli BL21(DE3) after temperature upshift from 37°C to 42°C, tiling arrays

Project description:Heat-responsive and time-resolved changes in transcriptome of E. coli BL21(DE3) Experimentally mapped transcriptome structure of Escherichia coli BL21(DE3) by hybridizing total RNA (including RNA species <200 nt) to genome-wide high-density tiling arrays (60 mer probes tiled every 10 nt).

2020-04-12 | GSE148034 | GEO

Microaerobic fermentation alters lactose metabolism in Escherichia coli

Project description:We report the effect of oxygenation state in lactose grown escherichia coli producing recombinant proteins. To shed more light on the mechanistic correlation between the uptake of lactose and dissolved oxygen, a comprehensive study has been undertaken with the E. coli BL21 (DE3) strain. Differences in consumption pattern of lactose, metabolites, biomass and product formation due to aerobiosis have been investigated. Transcriptomic profiling of metabolic changes due to aerobic process and microaerobic process during protein formation phase has been studied and the results provide a deeper understanding of protein production in E. coli BL21 (DE3) strains with lactose based promoter expression systems.This study also provides a scientific understanding of escherichia coli metabolism upon oxygen fluctuations.

2020-05-18 | GSE135698 | GEO

Deciphering the transcriptional changes in Escherichia coli strains C41(DE3) and C43(DE3) that makes them a superior choice for membrane protein production.

Project description:This study assessed the differential gene expression of C41(DE3) and C43(DE3) strains in comparison to their parental strain BL21(DE3), in the presence and absence of a prototypical protein expression vector with or without the inducer IPTG.

2022-06-23 | GSE153028 | GEO

DNA-seq of E. coli BL21 (DE3) Evolved in High pH (pH = 9) Terrific Broth Media

Project description:Whole genome sequencing was performed on E. coli BL21 (DE3) evolved at 25°C in pH 9 terrific broth media buffered with Tris-HCl (pH 9). The evolved E. coli was characterized and compared to the parent strain.

2023-10-31 | E-MTAB-12334 | biostudies-arrayexpress

RNA-seq of E. coli BL21 (DE3) Evolved in High pH (pH = 9) Terrific Broth Media

Project description:Bulk RNA-sequencing was performed on E. coli BL21 (DE3) evolved at 25°C in pH 9 terrific broth media buffered with Tris-HCl (pH 9). The evolved E. coli was characterised and compared to the parent strain during protein expression; the strains were actively grown and compared for gene expression at pH 7 and pH 9 in terrific broth media.

2023-10-31 | E-MTAB-12337 | biostudies-arrayexpress

Confirmation of recombinant Calcium-activated potassium channel from Leishmania donovani

Project description:Calcium-activated potassium channels are important membrane proteins that help in maintaining the cellular physiology and cell signalling. Till date no Calcium-activated potassium channels have been identified in Leishmania donovani. It’s gene was cloned into pET vector and expressed in E. coli BL21 (DE3). The recombinant protein was purified using nickel affinity chromatography.

2021-05-11 | PXD019577 | Pride

Time series of Escherichia coli BL21(DE3) growth curve, tiling arrays

Project description:Experimentally mapped transcriptome structure of Escherichia coli BL21(DE3) by hybridizing total RNA (including RNA species <200 nt) to genome-wide high-density tiling arrays (60 mer probes tiled every 10 nt).

2017-01-13 | GSE93565 | GEO

Transcription profiling by array of E. coli BL21(DE3)::TN7T7 -SOD to analyze the cellular response of this plasmid-free expression system to high level expression of recombinant human super-oxide-dismutase

Project description:This transcription profiling time course experiment was conducted in order to analyze the cellular response of the plasmid-free expression system E.coli BL21(DE3)::TN7<T7 -SOD> to high level expression of recombinant human super-oxide-dismutase (SOD).Three biological replicates were generated by using a carbon limited exponential fed-batch cultivation similar to industrial setups for large scale production. For induction of the system a single pulse of isopropyl-beta-D-galactoside (IPTG) yielding in a fully induced system is applied one doubling past feed start.

2013-05-31 | E-MARS-24 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data