Temporal quantification of MAPK induced expression in single yeast cells.

Détails

ID Serval
serval:BIB_C5D261D6B9AB
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Temporal quantification of MAPK induced expression in single yeast cells.
Périodique
Journal of Visualized Experiments
Auteur⸱e⸱s
Pelet S., Aymoz D., Durandau E.
ISSN
1940-087X (Electronic)
ISSN-L
1940-087X
Statut éditorial
Publié
Date de publication
2013
Peer-reviewed
Oui
Numéro
80
Pages
1-8
Langue
anglais
Résumé
The quantification of gene expression at the single cell level uncovers novel regulatory mechanisms obscured in measurements performed at the population level. Two methods based on microscopy and flow cytometry are presented to demonstrate how such data can be acquired. The expression of a fluorescent reporter induced upon activation of the high osmolarity glycerol MAPK pathway in yeast is used as an example. The specific advantages of each method are highlighted. Flow cytometry measures a large number of cells (10,000) and provides a direct measure of the dynamics of protein expression independent of the slow maturation kinetics of the fluorescent protein. Imaging of living cells by microscopy is by contrast limited to the measurement of the matured form of the reporter in fewer cells. However, the data sets generated by this technique can be extremely rich thanks to the combinations of multiple reporters and to the spatial and temporal information obtained from individual cells. The combination of these two measurement methods can deliver new insights on the regulation of protein expression by signaling pathways.
Pubmed
Création de la notice
08/01/2014 17:39
Dernière modification de la notice
20/08/2019 15:41
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