| Autor: |
Greco FV; School of Biological Sciences, University of Bristol, Bristol, UK., Pandi A; Department of Biochemistry and Synthetic Metabolism, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany., Erb TJ; Department of Biochemistry and Synthetic Metabolism, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.; SYNMIKRO Center of Synthetic Microbiology, Marburg, Germany., Grierson CS; School of Biological Sciences, University of Bristol, Bristol, UK.; BrisSynBio, University of Bristol, Bristol, UK., Gorochowski TE; School of Biological Sciences, University of Bristol, Bristol, UK. thomas.gorochowski@bristol.ac.uk.; BrisSynBio, University of Bristol, Bristol, UK. thomas.gorochowski@bristol.ac.uk. |
| Abstrakt: |
Strictly controlled inducible gene expression is crucial when engineering biological systems where even tiny amounts of a protein have a large impact on function or host cell viability. In these cases, leaky protein production must be avoided, but without affecting the achievable range of expression. Here, we demonstrate how the central dogma offers a simple solution to this challenge. By simultaneously regulating transcription and translation, we show how basal expression of an inducible system can be reduced, with little impact on the maximum expression rate. Using this approach, we create several stringent expression systems displaying >1000-fold change in their output after induction and show how multi-level regulation can suppress transcriptional noise and create digital-like switches between 'on' and 'off' states. These tools will aid those working with toxic genes or requiring precise regulation and propagation of cellular signals, plus illustrate the value of more diverse regulatory designs for synthetic biology. |
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