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.
Project description:We measured the mRNA abundance in E.coli using RNAseq to calculate mRNA lifetimes. The data is used in support of a larger paper on the proteome and transcriptome of E.coli.
Project description:This transcription profiling time course experiment was conducted in order to analyze the cellular response of the T7 RNA polymerase based E. coli expression system HMS174(DE3)(pET30aNproGFP) to a limited induction with isopropyl-beta-D-galactoside (IPTG). The used cell material was produced under well controlled and defined conditions in an exponential carbon limited fed-batch cultivation at a growth rate of 0.1 per hour. One doubling past feed start induction was started by continuously feeding limiting amounts of inducer gaining in a constant IPTG to cell dry weight ratio of 1µmol/g. Due to the low induction level a physiologically tolerable recombinant gene expression rate was generated and production period was significantly elongated. Non induced cells sampled at the time point of induction were used as reference and compared to cells subsequently sampled during the production period (sampling frequency 2 hours).
Project description:We measured the mRNA abundance in E.coli using RNAseq to calculate mRNA lifetimes. The data is used in support of a larger paper on the proteome and transcriptome of E.coli. Comparison of mRNA abundance over time, after the addition of transcription inhibitor, rifampicin. Center: Harvard University
Project description:Escherichia coli release Extracellular Vesicles (EVs) which carry diverse molecular cargo. Pathogenic E.coli EVs contain virulence factors which assist during infection in the host in different mechanisms.The RNA cargo of E.coli EVs has not been assessed in their effect in the host. We used microarray data to asses and compare the global response of bladder cells to EV-RNA from pathogenic E.coli (Uropathogenic UPEC 536) and non-pathogenic E. coli (probiotic Nissle 1917)
Project description:This experiment studied the effect of FPP accumulation on E. coli. E. coli cells transformed with pMBIS (the S. cerevisiae mevalonate pathway enzymes converting mevalonate to FPP) and fed mevalonate produce large amounts of FPP, which causes toxicity when it accumulates. When coupled with an active amorphadiene synthase (pADS) the cells produce amorphadiene, a non-toxic isoprenoid. To accumulate FPP, but maintain similar protein burden, an amorphadiene synthase with 3 mutations to render it inactive was used (pADSmut) to accumulate FPP. E. coli was transformed with pMBIS and pADS or pMBIS and pADSMut and grown in M9+glucose with varying magnesium concentrations and fed 20 mM mevalonate and induced with 0.5 mM IPTG, then sampled at subsequent time points.
Project description:This experiment studied the effect of FPP accumulation on E. coli. E. coli cells transformed with pMBIS (the S. cerevisiae mevalonate pathway enzymes converting mevalonate to FPP) and fed mevalonate produce large amounts of FPP, which causes toxicity when it accumulates. When coupled with an active amorphadiene synthase (pADS) the cells produce amorphadiene, a non-toxic isoprenoid. To accumulate FPP, but maintain similar protein burden, an amorphadiene synthase with 3 mutations to render it inactive was used (pADSmut) to accumulate FPP. E. coli was transformed with pMBIS and pADS or pMBIS and pADSMut and grown in LB and fed 10 mM mevalonate and induced with 0.5 mM IPTG, then sampled at subsequent time points.
