| Value |
12
Min
Range: Table - link Min
|
| Organism |
Generic |
| Reference |
Iizuka R, Yamagishi-Shirasaki M, Funatsu T. Kinetic study of de novo chromophore maturation of fluorescent proteins. Anal Biochem. 2011 Jul 15 414(2):173-8. p.176 table 1PubMed ID21459075
|
| Method |
(P.175 left column bottom paragraph:) "The PURE system is a coupled cell-free transcription/ translation system reconstituted from the purified components necessary for translation in E. coli [refs 14,15]." (P.175 right column top paragraph:) "The maturation kinetics was slow enough to monitor in real time with a spectrofluorometer and fitted well to a single exponential curve, suggesting that the oxidation step is the rate-limiting step in the overall chromophore maturation process (Figs. 1 and 2 see also supplementary material). The observed rate constant was defined as the rate constant of de novo chromophore maturation." |
| Comments |
"During earlier years, the maturation rate of GFP was determined as the rate of fluorescence development after admission of air to anaerobically expressed GFP [ref 5]. Currently, conventional methods to study the maturation kinetics are based on triggering protein folding of urea-solubilized inclusion bodies [ref 9] and reoxidation of the chromophore after chemical reduction of the mature chromophore [refs 9–13]. However, inconsistent data have been reported (Table 1)." |
| Entered by |
Uri M |
| ID |
106893 |