article · 15/10/2018
The inducible chemical-genetic fluorescent marker FAST outperforms classical fluorescent proteins in the quantitative reporting of bacterial biofilm dynamics
Résumé
To increase our understanding of bacterial biofilm complexity, real- time quantitative analyses of the living community functions are required. To reach this goal, accurate fluorescent reporters are needed. In this paper, we used the classical fluorescent genetic reporters of the GFP family and demonstrated their limits in the context of a living biofilm. We showed that fluorescence signal saturated after only a few hours of growth and related this saturation to the reduction of oxygen concentration induced by bacterial consumption. This behaviour prevents the use of GFP-like fluorescent proteins for quantitative measurement in living biofilms. To overcome this limitation, we propose the use of a recently introduced small protein tag, FAST, which is fluorescent in the presence of an exogenously applied fluorogenic dye, enabling to avoid the oxygen sensitivity issue. We compared the ability of FAST to report on biofilm growth with that of GFP and mCherry, and demonstrated the superiority of the FAST:fluorogen probes for investigating dynamics in the complex environment of a living biofilm.
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Monmeyran, A., Thomen, P., Jonquiere, H., Sureau, F., Li, C., Plamont, M.-A., Douarche, C., Casella, J.-F., Gautier, A., & Henry, N. (2018). The inducible chemical-genetic fluorescent marker FAST outperforms classical fluorescent proteins in the quantitative reporting of bacterial biofilm dynamics. Scientific Reports, 8(1), 10336. https://doi.org/10.1038/s41598-018-28643-z@article{Monmeyran2018_290,
author = {Monmeyran, Amaury and Thomen, Philippe and Jonquiere, Hugo and Sureau, Franck and Li, Chenge and Plamont, Marie-Aude and Douarche, Carine and Casella, Jean-François and Gautier, Arnaud and Henry, Nelly},
year = {2018},
month = {10},
title = {The inducible chemical-genetic fluorescent marker FAST outperforms classical fluorescent proteins in the quantitative reporting of bacterial biofilm dynamics},
journal = {Scientific Reports},
volume = {8},
number = {1},
pages = {10336},
abstract = {To increase our understanding of bacterial biofilm complexity, real- time quantitative analyses of the living community functions are required. To reach this goal, accurate fluorescent reporters are needed. In this paper, we used the classical fluorescent genetic reporters of the GFP family and demonstrated their limits in the context of a living biofilm. We showed that fluorescence signal saturated after only a few hours of growth and related this saturation to the reduction of oxygen concentration induced by bacterial consumption. This behaviour prevents the use of GFP-like fluorescent proteins for quantitative measurement in living biofilms. To overcome this limitation, we propose the use of a recently introduced small protein tag, FAST, which is fluorescent in the presence of an exogenously applied fluorogenic dye, enabling to avoid the oxygen sensitivity issue. We compared the ability of FAST to report on biofilm growth with that of GFP and mCherry, and demonstrated the superiority of the FAST:fluorogen probes for investigating dynamics in the complex environment of a living biofilm.},
url = {https://www.ncbi.nlm.nih.gov/pubmed/29985417},
doi = {10.1038/s41598-018-28643-z},
issn = {2045-2322},
}TY - JOUR
AU - Monmeyran, Amaury
AU - Thomen, Philippe
AU - Jonquiere, Hugo
AU - Sureau, Franck
AU - Li, Chenge
AU - Plamont, Marie-Aude
AU - Douarche, Carine
AU - Casella, Jean-François
AU - Gautier, Arnaud
AU - Henry, Nelly
PY - 2018
DA - 2018/10/15
TI - The inducible chemical-genetic fluorescent marker FAST outperforms classical fluorescent proteins in the quantitative reporting of bacterial biofilm dynamics
JO - Scientific Reports
VL - 8
IS - 1
SN - 2045-2322
AB - To increase our understanding of bacterial biofilm complexity, real- time quantitative analyses of the living community functions are required. To reach this goal, accurate fluorescent reporters are needed. In this paper, we used the classical fluorescent genetic reporters of the GFP family and demonstrated their limits in the context of a living biofilm. We showed that fluorescence signal saturated after only a few hours of growth and related this saturation to the reduction of oxygen concentration induced by bacterial consumption. This behaviour prevents the use of GFP-like fluorescent proteins for quantitative measurement in living biofilms. To overcome this limitation, we propose the use of a recently introduced small protein tag, FAST, which is fluorescent in the presence of an exogenously applied fluorogenic dye, enabling to avoid the oxygen sensitivity issue. We compared the ability of FAST to report on biofilm growth with that of GFP and mCherry, and demonstrated the superiority of the FAST:fluorogen probes for investigating dynamics in the complex environment of a living biofilm.
SP - 10336
DO - 10.1038/s41598-018-28643-z
UR - https://www.ncbi.nlm.nih.gov/pubmed/29985417
ER -