Projects per year
Abstract
The ability to induce gene expression in a small molecule dependent manner has led to many applications in target discovery, functional elucidation and bio-production. To date these applications have relied on a limited set of protein-based control mechanisms operating at the level of transcription initiation. The discovery, design and reengineering of riboswitches offer an alternative means by which to control gene expression. Here we report the development and characterization of a novel tunable recombinant expression system, termed RiboTite, which operates at both the transcriptional and translational level. Using standard inducible promoters and orthogonal riboswitches, a multi-layered modular genetic control circuit was developed to control the expression of both bacteriophage T7 RNA polymerase and recombinant gene(s) of interest. The system was benchmarked against a number of commonly used E. coli expression systems, and shows tight basal control, precise analogue tunability of gene expression at the cellular level, dose-dependent regulation of protein production rates over extended growth periods and enhanced cell viability. This novel system expands the number of E. coli expression systems for use in recombinant protein production and represents a major performance enhancement over and above the most widely used expression systems.
Original language | English |
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Article number | e21 |
Journal | Nucleic acids research |
Volume | 44 |
Issue number | 3 |
Early online date | 23 Sept 2015 |
DOIs | |
Publication status | Published - 18 Feb 2016 |
Research Beacons, Institutes and Platforms
- Manchester Institute of Biotechnology
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Dive into the research topics of 'Dual transcriptional-translational cascade permits cellular level tuneable expression control.'. Together they form a unique fingerprint.Projects
- 3 Finished
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Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals
Scrutton, N., Azapagic, A., Balmer, A., Barran, P., Breitling, R., Delneri, D., Dixon, N., Faulon, J., Flitsch, S., Goble, C., Goodacre, R., Hay, S., Kell, D., Leys, D., Lloyd, J., Lockyer, N., Martin, P., Micklefield, J., Munro, A., Pedrosa Mendes, P., Randles, S., Salehi Yazdi, F., Shapira, P., Takano, E., Turner, N. & Winterburn, J.
14/11/14 → 13/05/20
Project: Research
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Development and Application of Next Generation Synthetic Biology Tools
1/11/13 → 31/10/19
Project: Research
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Linking experiment to theory: Quantum entanglement during enzyme catalysis - Dr S Hay fellowship
1/09/10 → 31/08/15
Project: Research