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Comparative transcriptional profiling of the bacterial stress response in temperature and chemically-induced recombinant E. coli processes

Background

Production of heterologous proteins results in a number of metabolic and physiological changes in the host cells during the course of a production process, namely the induction of stress responses and corresponding alterations in gene expression profiles [1].

Results

This study focuses on quantitative monitoring of the adaptation of E. coli to recombinant protein production on the transcriptome level by a bead-based RNA sandwich hybridisation assay, a rapid novel method based on the detection of hybridisation events between specific oligonucleotide probes and the target nucleic acids [2, 3].

The expression profiles of selected genes including the product gene, anabolic and stress responsive genes were quantitatively analyzed in cells producing the human basic fibroblast growth factor (hFGF-2), a protein that partially aggregates into inclusion bodies. Transcriptome profiles during temperature- and IPTG-induced synthesis of hFGF-2 using the K12 strain TG1 and BL21(DE3) as production hosts, respectively, were compared.

References

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    Hoffmann F, Rinas U: Stress induced by recombinant protein production in Escherichia coli. Adv Biochem Eng Biotechnol. 2004, 89: 73-92.

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    Gabig-Ciminska M, Holmgren A, Andresen H, Barken K, Wümpelmann M, Albers J, Hintsche R, Breitenstein A, Neubauer P, Los M, et al.: Electric chips for rapid detection and quantification of nucleic acids. Biosens Bioelectron. 2004, 19: 537-546. 10.1016/S0956-5663(03)00273-2.

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    Soini J, Falschlehner C, Mayer C, Böhm D, Panula J, Vasala A, Neubauer P: Transient increase of ATP as a response to temperature up-shift in Escherichia coli. Microb Cell Fact. 2005, 4: 9- 10.1186/1475-2859-4-9.

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Correspondence to Ursula Rinas.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution 2.0 International License (https://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Böhm, D., Rinas, U. Comparative transcriptional profiling of the bacterial stress response in temperature and chemically-induced recombinant E. coli processes. Microb Cell Fact 5, P1 (2006). https://doi.org/10.1186/1475-2859-5-S1-P1

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Keywords

  • Basic Fibroblast Growth Factor
  • Stress Responsive Gene
  • Recombinant Protein Production
  • Hybridisation Event
  • Coli Process