ECFP maturation time

Value 49 min Range: ±9 min
Organism Budding yeast Saccharomyces cerevisiae
Reference Gordon A, Colman-Lerner A, Chin TE, Benjamin KR, Yu RC, Brent R. Single-cell quantification of molecules and rates using open-source microscope-based cytometry. Nat Methods. 2007 Feb4(2):175-81. p.178 right column 3rd paragraphPubMed ID17237792
Method P.178 right column 2nd paragraph: "[Researchers] quantified maturation time [refs 22,23] for YFP (wild-type GFP with mutations S65G,V68L,S72A,T203Y) and a cyan fluorescent protein derivative (CFP wild-type GFP with mutations F64,S65T,Y66W,N146I,M153T,V163A), in individual S. cerevisiae at 25°C (ref. 3 Supplementary Note). [They] induced fluorescent protein synthesis by addition of pheromone to cells with integrated constructs in which the fluorescent protein replaced the PRM1 ORF, allowed transcription and translation to proceed for 30 min, then added cycloheximide to block translation. Cells reached maximum fluorescence by 3 h after translation stop (Fig. 4a) and photobleaching-corrected fluorescence (Supplementary Note) remained stable for up to 24 h (data not shown), indicating that under these conditions protein degradation was negligible."
Comments P.178 right column 3rd paragraph: "Although individual cells varied in the total amount of YFP by a factor of four (Fig. 4a), there was little cell-to-cell variation in the maturation rates (Fig. 4b coefficient of variation o 0.1 Supplementary Note). [Researchers] performed an identical analysis for CFP (data not shown). In yeast under these conditions, YFP and CFP form mature fluorophores at similar rates, with average halftimes for maturation of 39 ± 7 and 49 ± 9 min, respectively (Supplementary Note)."
Entered by Uri M
ID 106883