Probing chemotaxis activity in Escherichia coli using fluorescent protein fusions.

Détails

Ressource 1Télécharger: s41598-019-40655-x.pdf (6890.56 [Ko])
Etat: Public
Version: Final published version
Licence: CC BY 4.0
ID Serval
serval:BIB_73F38E4B18B0
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Probing chemotaxis activity in Escherichia coli using fluorescent protein fusions.
Périodique
Scientific Reports
Auteur(s)
Roggo C., Carraro N., van der Meer J.R.
ISSN
2045-2322 (Electronic)
ISSN-L
2045-2322
Statut éditorial
Publié
Date de publication
2019
Peer-reviewed
Oui
Volume
9
Numéro
1
Pages
3845
Langue
anglais
Résumé
Bacterial chemotaxis signaling may be interesting for the development of rapid biosensor assays, but is difficult to quantify. Here we explore two potential fluorescent readouts of chemotactically active Escherichia coli cells. In the first, we probed interactions between the chemotaxis signaling proteins CheY and CheZ by fusing them individually with non-fluorescent parts of stable or unstable 'split'-Green Fluorescent Protein. Wild-type chemotactic cells but not mutants lacking the CheA kinase produced distinguishable fluorescence foci, two-thirds of which localize at the cell poles with the chemoreceptors and one-third at motor complexes. Fluorescent foci based on stable split-eGFP displayed small fluctuations in cells exposed to attractant or repellent, but those based on an unstable ASV-tagged eGFP showed a higher dynamic behaviour both in the foci intensity changes and the number of foci per cell. For the second readout, we expressed the pH-sensitive fluorophore pHluorin in the cyto- and periplasm of chemotactically active E. coli. Calibrations of pHluorin fluorescence as a function of pH demonstrated that cells accumulating near a chemo-attractant temporally increase cytoplasmic pH while decreasing periplasmic pH. Both readouts thus show promise for biosensor assays based on bacterial chemotaxis activity.
Pubmed
Web of science
Open Access
Oui
Création de la notice
25/03/2019 18:23
Dernière modification de la notice
20/08/2019 15:31
Données d'usage