Construction of a Bioassay System to Identify Extracellular Agents Targeting Bacterial Cell Envelope.
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.
Department of Biological and Medical Sciences, Faculty of Health and Life Sciences, Oxford Brookes University, Headington, Oxford, OX3 0BP, UK. hee-jeon.hong@brookes.ac.uk.|Department of Biochemistry, University of Cambridge, Cambridge, UK. hee-jeon.hong@brookes.ac.uk.
Hee-Jeon
HJ
Hong
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001681:Biological Assay; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism*; D024881:Drug Resistance, Bacterial; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D017930:Genes, Reporter; D007612:Kanamycin / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012808:Sigma Factor / Q000235:genetics* / Q000378:metabolism; D013291:Streptococcus / Q000166:cytology / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D012808:Sigma Factor; D007612:Kanamycin
Bioassay system; Cell wall stress; Streptomyces coelicolor; Transcriptional fusion; sigE