A novel mutant of green fluorescent protein with enhanced sensitivity for microanalysis at 488 nm excitation.
Green fluorescent protein (GFP) has been utilized as a powerful reporter of gene expression and protein localization in cells. We discovered a mutant carrying point mutation S208L from a UV-excitable GFP (F99S/M153T/V163A). It had the enhanced fluorescence intensity. Introduction of the red-shifted mutations (F64L/S65T) to this mutant led to the GFP having the brightest mutants reported which were expressed in Escherichia coli and excited at 488 nm. The relative fluorescence intensities to that of wild-type GFP and GFPuv were increased about 120- and 10-fold, respectively. It was shown that the S208L mutation contributes to both a higher intrinsic brightness of GFP and a higher expression level in E. coli.
Ito
Y
Y
Department of Functional Materials Science, Saitama University, Urawa, 338-8570, Japan.
Suzuki
M
M
Husimi
Y
Y
D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005453:Fluorescence; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids; D017354:Point Mutation; D013050:Spectrometry, Fluorescence
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins