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Streptomyces venezuelae TX-TL - a next generation cell-free synthetic biology tool

Moore, Simon J., Lai, Hung-En, Needham, Hannah, Polizzi, Karen M., Freemont, Paul S. (2017) Streptomyces venezuelae TX-TL - a next generation cell-free synthetic biology tool. Biotechnology Journal, 12 (4). ISSN 1860-6768. (doi:10.1002/biot.201600678) (KAR id:69452)

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Abstract

Streptomyces venezuelae is a promising chassis in synthetic biology for fine chemical and secondary metabolite pathway engineering. The potential of S. venezuelae could be further realized by expanding its capability with the introduction of its own in vitro transcription-translation (TX-TL) system. TX-TL is a fast and expanding technology for bottom-up design of complex gene expression tools, biosensors and protein manufacturing. Herein, we introduce a S. venezuelae TX-TL platform by reporting a streamlined protocol for cell-extract preparation, demonstrating high-yield synthesis of a codon-optimized sfGFP reporter and the prototyping of a synthetic tetracycline-inducible promoter in S. venezuelae TX-TL based on the TetO-TetR repressor system. The aim of this system is to provide a host for the homologous production of exotic enzymes from Actinobacteria secondary metabolism in vitro. As an example, we demonstrate the soluble synthesis of a selection of enzymes (12-70 kDa) from the Streptomyces rimosus oxytetracycline pathway.

Item Type: Article
DOI/Identification number: 10.1002/biot.201600678
Uncontrolled keywords: Cell-free, gene expression, Streptomyces, synthetic biology, tetracycline
Subjects: Q Science
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Simon Moore
Date Deposited: 09 Oct 2018 09:55 UTC
Last Modified: 16 Jan 2020 14:28 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/69452 (The current URI for this page, for reference purposes)
Moore, Simon J.: https://orcid.org/0000-0002-1968-206X
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