GFP (S65T) is a basic (constitutively fluorescent) green fluorescent protein published in 1995, derived from Aequorea victoria.

We chose to work on GFP/S65T for two reasons. First, GFP/S65T is primarily excited in the blue region of the visible spectrum, rather than in the UV as with wild-type and is therefore more useful for imaging applications.

An engineered sGFP(S65T) sequence containing optimized codons of highly expressed eukaryotic proteins has provided up to 100-fold brighter fluorescence signals than the original jellyfish GFP sequence in plant and mammalian cells.

The green fluorescent protein (GFP) was used as a noninvasive probe to quantify the rheological properties of cell cytoplasm. GFP mutant S65T was purified from recombinant bacteria for solution studies, and expressed in CHO cell cytoplasm.

In 1995, Roger Y. Tsien described an S65T point mutation that increased the fluorescence intensity and photostability of GFP. This also shifted its major excitation peak from 395 nm to 488 nm, effectively ameliorating the deficiencies found in the wildtype protein and facilitating its widespread use in research.

The double mutant GFP S65T/H148D is unusual with respect to a number of its photo- and biophysical properties. The introduction of the S65T mutation into wt-GFP leads to loss of fluorescence upon excitation of the A band, which is restored by the second site mutation H148D.

Plasmid pFA6a-GFP (S65T)-kanMX6 from Dr. John Pringle's lab contains the insert KanR and is published in Yeast. 1998 Jul;14 (10):943-51. This plasmid is available through Addgene.

Apr 10, 1999 · To explore the structural basis for the titration behavior of the popular GFP S65T variant, we determined high-resolution crystal structures at pH 8.0 and 4.6. The structures revealed changes in the hydrogen bond pattern with the chromophore, suggesting that the pH sensitivity derives from protonation of the chromophore phenolate.

Jul 7, 2007 · We have generated the mutant S65T/G67A-GFP, which is unable to form the cyclic chromophore, with the goal of investigating the folding, unfolding and competing aggregation of GFP under fully reversible conditions.

Apr 1, 1997 · GFP mutant S65T was purified from recombinant bacteria for solution studies, and expressed in CHO cell cytoplasm. GFP-S65T was brightly fluorescent in solution (lambda ex 492 nm, lambda em 509 nm) with a lifetime of 2.9 ns and a rotational correlation time (tc) of 20 ns.