Abstract
sigE in Streptomyces coelicolor encodes an extracytoplasmic function (ECF) sigma factor, σ E, which is part of a signal transduction system that senses and responds to general cell wall stress in S. coelicolor. Expression of sigE is induced by a wide variety of agents that stress the cell wall under the control of two-component signal transduction system, CseB/CseC encoded in the same operon where sigE was identified from. Here we describe a method developing a bioassay system in S. coelicolor via a transcriptional fusion in which the promoter of sigE operon and a reporter gene (neo) conferring resistance to kanamycin were used. The effectiveness of the resulting bioassay system was determined by monitoring various agents that cause bacterial cell wall stress such as lysozyme or some antibiotics that target cell wall. In consequence, the result confirms that the bioassay system has a potential to be a simple but effective screening tool for identifying novel extracellular agents targeting bacterial cell wall.
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Acknowledgments
This work was supported by funding from the Medical Research council, UK (G0700141) and the Royal Society, UK (516002.K5877/ROG). The author would like to thank Andy Hesketh (University of Cambridge, Systems Biology Centre) for comments on the manuscript.
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Hong, HJ. (2016). Construction of a Bioassay System to Identify Extracellular Agents Targeting Bacterial Cell Envelope. In: Hong, HJ. (eds) Bacterial Cell Wall Homeostasis. Methods in Molecular Biology, vol 1440. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3676-2_10
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DOI: https://doi.org/10.1007/978-1-4939-3676-2_10
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