Buckley, A. M., Jukes, C., Candlish, D., Irvine, J. J., Spencer, J., Fagan, R. P., Roe, A. J. , Christie, J. , Fairweather, N. F. and Douce, G. R. (2016) Lighting up Clostridium Difficile: reporting gene expression using fluorescent Lov domains. Scientific Reports, 6, 23463. (doi: 10.1038/srep23463) (PMID:26996606) (PMCID:PMC4800718)
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Abstract
The uses of fluorescent reporters derived from green fluorescent protein have proved invaluable for the visualisation of biological processes in bacteria grown under aerobic conditions. However, their requirement for oxygen has limited their application in obligate anaerobes such as Clostridium difficile. Fluorescent proteins derived from Light, Oxygen or Voltage sensing (LOV) domains have been shown to bridge this limitation, but their utility as translational fusions to monitor protein expression and localisation in a strict anaerobic bacterium has not been reported. Here we demonstrate the utility of phiLOV in three species of Clostridium and its application as a marker of real-time protein translation and dynamics through genetic fusion with the cell division protein, FtsZ. Time lapse microscopy of dividing cells suggests that Z ring assembly arises through the extension of the FtsZ arc starting from one point on the circumference. Furthermore, through incorporation of phiLOV into the flagella subunit, FliC, we show the potential of bacterial LOV-based fusion proteins to be successfully exported to the extracellular environment.
Item Type: | Articles |
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Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Roe, Professor Andrew and Spencer, Dr Janice and Douce, Dr Gillian and Buckley, Dr Anthony and Christie, Professor John and Candlish, Mrs Denise |
Authors: | Buckley, A. M., Jukes, C., Candlish, D., Irvine, J. J., Spencer, J., Fagan, R. P., Roe, A. J., Christie, J., Fairweather, N. F., and Douce, G. R. |
College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity |
Journal Name: | Scientific Reports |
Publisher: | Nature Publishing Group |
ISSN: | 2045-2322 |
ISSN (Online): | 2045-2322 |
Published Online: | 21 March 2016 |
Copyright Holders: | Copyright © 2016 The Authors |
First Published: | First published in Scientific Reports 6:23463 |
Publisher Policy: | Reproduced under a creative commons license |
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