Maturation time of tdTomato fluorescent protein

Value 1 Hour Range: Table - link Hour
Organism Bacteria Escherichia coli
Reference Shaner NC, Steinbach PA, Tsien RY. A guide to choosing fluorescent proteins. Nat Methods. 2005 Dec2(12):905-9. Article link - link Supplementary table 1 PubMed ID16299475
Primary Source Shaner NC et al., Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol. 2004 Dec22(12):1567-72.PubMed ID15558047
Method (Primary source:) "RFPs were expressed from pBAD vectors in E. coli LMG194 by growing single colonies in 40 ml RM/ampicillin supplemented with 0.2% D-glucose for 8 h, adding 40 ml LB/ampicillin and L-arabinose to a final concentration of 0.2%, and incubating overnight at 37°C. For maturation experiments, flasks were sealed with parafilm upon induction to restrict oxygen availability. All proteins were purified by Ni-NTA chromatography (Qiagen) and dialyzed into PBS. Biochemical and fluorescence characterization experiments were done as described [ref 5]. For FRET measurements, purified Zn2+ sensor proteins were diluted in 10 mM MOPS, 100 mM KCl, pH 7.4 with either 1 mM EDTA or 1 mM ZnCl2, and fluorescence emission spectra were collected with excitation near the peak donor excitation wavelength."
Entered by Uri M
ID 106876