The introduction of several mutations resulted in the generation of improved mutants of the green fluorescent protein (GFP). A strong green (GFPsg25) and blue (BFPsg50) fluorescent protein, gave 50-fold-100-fold brighter fluorescence compared to wild-type GFP and BFP (Tyr66His), respectively, upon expression in mammalian cells. GFPsg25 and BFPsg50 have different excitation and emission maxima. ...

background: baculovirus expression system is one of the most attractive and powerful eukaryotic expression systems for the production of recombinant proteins. the presence of a biomarker is required to monitor transfection efficiency or protein expression levels in insect cells. methods: the aim of this study was to construct a baculovirus expression vector encoding a copepod super green fluore...

Discovery of Green Fluorescent Protein (GFP) constituted an important improvement for living cell studies on submicron resolution allowing in vivo fluorescence labeling. We studied the photo-physical properties of single GFP molecules incorporated in a charged polyelectrolyte environment by means of single molecule spectroscopy. The fluorescence characteristics change dramatically in terms of p...

To investigate the role of protein kinase C (PKC) isoforms in regulation of neurite outgrowth, PKC a , b II, d , and e fused to enhanced green fluorescent protein (EGFP) were transiently overexpressed in neuroblastoma cells. Overexpression of PKC e –EGFP induced cell processes whereas the other isoforms did not. The effect of PKC e –EGFP was not suppressed by the PKC inhibitor GF109203X. Instea...

Fluorescent Proteins (FP) can be applied as biomarkers and biosensors in the industrial medical fields, but their large-scale use, especially for new applications, is limited due to low stability. Hence, discovery of additives capable preserving activity FP at room temperature under stress conditions help expand facilitate commercial use. With this goal, we evaluated application 1 - alkyl-3-met...

Fluorescent proteins (FPs) are indispensable markers for two-photon imaging of live tissue, especially in the brains of small model organisms. The quantity of physiologically relevant data collected, however, is limited by heat-induced damage of the tissue due to the high intensities of the excitation laser. We seek to minimize this damage by developing FPs with improved brightness. Among FPs w...

The unusual membrane motor protein prestin is essential for mammalian hearing and for the survival of cochlear outer hair cells. While prestin has been demonstrated to be a homooligomer, by Western blot and FRET analyses, the stoichiometry of self association is unclear. Prestin, coupled to the enhanced green fluorescent protein, was synthesized and membrane targeted in human embryonic kidney c...

When aequorin is microinjected into cleavage-stage zebrafish embryos, it is largely used up by ~24 hours. Thus, it is currently not possible to image Ca(2+) signals from later stages of zebrafish development using this approach. We have, therefore, developed protocols to express apoaequorin, i.e., the protein component of aequorin, transiently in zebrafish embryos and then reconstitute intact a...

BACKGROUND In this study, Escherichia coli cells producing red fluorescent protein of Discosoma sp. (drFP583 DsRed) were investigated with flow cytometry by using 488 nm excitation. We also studied whether green fluorescent protein (GFP) and drFP583 could be detected simultaneously from a single bacterial cell. METHODS Plasmids pDsRed and pEGFP were used for the production of drFP583 and enha...