Half-life of GFPmut3* in Escherichia coli and Pseudomonas putida in vivo

Range >1 day
Organism bacteria
Reference Andersen JB, Sternberg C, Poulsen LK, Bjorn SP, Givskov M, Molin S. New Unstable Variants of Green Fluorescent Protein for Studies of Transient Gene Expression in Bacteria. Appl Environ Microbiol. 1998 Jun64(6):2240-6. P.2244 right column 2nd & 3rd paragraphsPubMed ID9603842
Method P.2244 right column 2nd paragraph: "In order to estimate the half-lives of the mature Gfp variants in vivo, [investigators] carried out quantitative fluorometer measurements of green fluorescence expressed by the strains derived from E. coli MV1190(λ-pir) or P. putida KT2442 (lacI q) encoding the variant Gfps (the same strains used for the colony investigations). The strains were grown in rich medium and shifted at mid-log phase to minimal medium without inducer. After the shift there was no increase in OD450 for 4 to 5 h, which means that protein synthesis (including that of Gfp) was negligible during this period. Fluorescence was measured at time intervals after the shift (Fig. 3). The stability of Gfpmut3* and the four tagged versions was estimated by using the relative green fluorescence value as a function of the time after the shift, represented as the slopes of the curves in Fig. 3."
Comments P.2244 right column 2nd & 3rd paragraphs: "Consequently, [investigators] conservatively estimated the in vivo half-life of mature Gfpmut3* to be more than 1 day. In P. putida KT2442 (lacI q) the green fluorescence derived from Gfpmut3* was similarly unaffected by the shift (Fig. 3B). Because of this, combined with the observation that colonies of this strain continue to be bright green fluorescent for more than a week (Fig. 2), [they] also consider the in vivo half-life of mature Gfpmut3* in P. putida to be more than 1 day."
Entered by Uri M
ID 105191