Bacterial host and reporter gene optimization for genetically encoded whole cell biosensors - UMR CNRS 5600 EVS / GeoSciences et Environnement / Ecole des Mines de Saint-Etienne - F42023
Article Dans Une Revue Environmental Science and Pollution Research Année : 2017

Bacterial host and reporter gene optimization for genetically encoded whole cell biosensors

Résumé

Whole-cell biosensors based on reporter genes allow detection of toxic metals in water with high selectivity and sensitivity under laboratory conditions; nevertheless, their transfer to a commercial inline water analyzer requires specific adaptation and optimization to field conditions as well as economical considerations. We focused here on both the influence of the bacterial host and the choice of the reporter gene by following the responses of global toxicity biosensors based on constitutive bacterial promoters as well as arsenite biosensors based on the arsenite-inducible P-ars promoter. We observed important variations of the bioluminescence emission levels in five different Escherichia coli strains harboring two different lux-based biosensors, suggesting that the best host strain has to be empirically selected for each new biosensor under construction. We also investigated the bioluminescence reporter gene system transferred into Deinococcus deserti, an environmental, desiccation- and radiation-tolerant bacterium that would reduce the manufacturing costs of bacterial biosensors for commercial water analyzers and open the field of biodetection in radioactive environments. We thus successfully obtained a cell survival biosensor and a metal biosensor able to detect a concentration as low as 100 nM of arsenite in D. deserti. We demonstrated that the arsenite biosensor resisted desiccation and remained functional after 7 days stored in air-dried D. deserti cells. We also report here the use of a new near-infrared (NIR) fluorescent reporter candidate, a bacteriophytochrome from the magnetotactic bacterium Magnetospirillum magneticum AMB-1, which showed a NIR fluorescent signal that remained optimal despite increasing sample turbidity, while in similar conditions, a drastic loss of the lux-based biosensors signal was observed.
Fichier principal
Vignette du fichier
Brutesco et al depot.pdf (634.91 Ko) Télécharger le fichier
Origine Fichiers produits par l'(les) auteur(s)
Loading...

Dates et versions

hal-01640038 , version 1 (23-04-2018)

Identifiants

Citer

Catherine Brutesco, Sandra Preveral, Camille Escoffier, Elodie C. T. Descamps, Elsa Prudent, et al.. Bacterial host and reporter gene optimization for genetically encoded whole cell biosensors. Environmental Science and Pollution Research, 2017, 24 (1), pp.52--65. ⟨10.1007/s11356-016-6952-2⟩. ⟨hal-01640038⟩
434 Consultations
390 Téléchargements

Altmetric

Partager

More