Br. Terry et al., MOLECULAR CHARACTERIZATION OF RECOMBINANT GREEN FLUORESCENT PROTEIN BY FLUORESCENCE CORRELATION MICROSCOPY, Biochemical and biophysical research communications, 217(1), 1995, pp. 21-27
The cDNA for the green fluorescent protein (GFP) of Aequorea victoria
has been expressed in transformed cells of Saccharomyces cerevisiae an
d the recombinant GFP isolated. Protonation and deprotonation of the c
loned and purified GFP produced major effects on its spectral absorpti
on characteristics with an increase in pH enhancing the fluorescence e
mission of the GFP more than twofold. Finally, molecular characterisat
ion of GFP by fluorescence correlation microscopy in a minimal target
volume of 1 fL yielded a translational diffusion coefficient (D-T) of
8.7x10(-7) cm(2).sec(-1), equivalent to a Stokes radius of 2.82nm for
a monodisperse globular protein of 27kDa. (C) Academic Press, Inc.