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Yeast

Title:		An improved short-lived fluorescent protein transcriptional reporter for Saccharomyces cerevisiae
Author(s):		Houser JR; Ford E; Chatterjea SM; Maleri S; Elston TC; Errede B;
Address:		"Department of Physics, University of North Carolina, Chapel Hill, NC 27599-7260, USA"
Journal Title:		Yeast
Year:		2012
Volume:		20121121
Issue:		12
Page Number:		519 - 530
DOI:		10.1002/yea.2932
ISSN/ISBN:		1097-0061 (Electronic) 0749-503X (Print) 0749-503X (Linking)
Abstract:		"Ideal reporter genes for temporal transcription programmes have short half-lives that restrict their detection to the window in which their transcripts are present and translated. In an effort to meet this criterion for reporters of transcription in individual living cells, we adapted the ubiquitin fusion strategy for programmable N-end rule degradation to generate an N-degron version of green fluorescent protein (GFP) with a half-life of ~7 min. The GFP variant we used here (designated GFP*) has excellent fluorescence brightness and maturation properties, which make the destabilized reporter well suited for tracking the induction and attenuation kinetics of gene expression in living cells. These attributes are illustrated by its ability to track galactose- and pheromone-induced transcription in S. cerevisiae. We further show that the fluorescence measurements using the short-lived N-degron GFP* reporter gene accurately predict the transient mRNA profile of the prototypical pheromone-induced FUS1 gene"
Keywords:		"Galactose/metabolism Gene Expression *Genes, Reporter Green Fluorescent Proteins/*genetics/metabolism Half-Life Kinetics Membrane Proteins/genetics Pheromones/metabolism Plasmids Promoter Regions, Genetic RNA, Fungal/genetics RNA, Messenger/genetics Sacch;"
Notes:		"MedlineHouser, John R Ford, Eintou Chatterjea, Sudeshna M Maleri, Seth Elston, Timothy C Errede, Beverly eng R01 GM079271/GM/NIGMS NIH HHS/ R01 GM084071/GM/NIGMS NIH HHS/ GM 084071/GM/NIGMS NIH HHS/ GM 079271/GM/NIGMS NIH HHS/ Research Support, N.I.H., Extramural England 2012/11/23 Yeast. 2012 Dec; 29(12):519-30. doi: 10.1002/yea.2932. Epub 2012 Nov 21"