Half-life measurements of chemical inducers for recombinant gene expression

Nicolo' Politi, Lorenzo Pasotti, Susanna Zucca, Michela Casanova, Giuseppina Micoli, Maria G. Cusella De Angelis, Paolo Magni

Research output: Contribution to journalArticle

Abstract

Background: Inducible promoters are widely spread genetic tools for triggering, tuning and optimizing the expression of recombinant genes in engineered biological systems. Most of them are controlled by the addition of a specific exogenous chemical inducer that indirectly regulates the promoter transcription rate in a concentration-dependent fashion. In order to have a robust and predictable degree of control on promoter activity, the degradation rate of such chemicals should be considered in many applications like recombinant protein production.Results: In this work, we use whole-cell biosensors to assess the half-life of three commonly used chemical inducers for recombinant Escherichia coli: Isopropyl β-D-1-thiogalactopyranoside (IPTG), anhydrotetracycline (ATc) and N-(3-oxohexanoyl)-L-homoserine lactone (HSL). A factorial study was conducted to investigate the conditions that significantly contribute to the decay rate of these inducers. Temperature has been found to be the major factor affecting ATc, while medium and pH have been found to highly affect HSL. Finally, no significant degradation was observed for IPTG among the tested conditions.Conclusions: We have quantified the decay rate of IPTG, ATc and HSL in many conditions, some of which were not previously tested in the literature, and the main effects affecting their degradation were identified via a statistics-based framework. Whole-cell biosensors were successfully used to conduct this study, yielding reproducible measurements via simple multiwell-compatible assays. The knowledge of inducer degradation rate in several contexts has to be considered in the rational design of synthetic biological systems for improving the predictability of induction effects, especially for prolonged experiments.

Original languageEnglish
Article number5
JournalJournal of Biological Engineering
Volume8
Issue number1
DOIs
Publication statusPublished - Feb 1 2014

Keywords

  • ATc
  • BioBrick™
  • Chemical inducers
  • Degradation rate
  • HSL
  • IPTG
  • Synthetic biology
  • Whole-cell biosensors

ASJC Scopus subject areas

  • Biomedical Engineering
  • Environmental Engineering
  • Cell Biology
  • Molecular Biology

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  • Cite this

    Politi, N., Pasotti, L., Zucca, S., Casanova, M., Micoli, G., Cusella De Angelis, M. G., & Magni, P. (2014). Half-life measurements of chemical inducers for recombinant gene expression. Journal of Biological Engineering, 8(1), [5]. https://doi.org/10.1186/1754-1611-8-5