Intraocular in vivo imaging of activated T-lymphocytes expressing green-fluorescent protein after stimulation with endotoxin

2001 ◽  
Vol 239 (8) ◽  
pp. 609-612 ◽  
Author(s):  
Matthias D. Becker ◽  
Sergio Crespo ◽  
Tammy M. Martin ◽  
Stephen R. Planck ◽  
Mayumi Naramura ◽  
...  
2007 ◽  
Vol 6 (2) ◽  
pp. 7290.2007.00007 ◽  
Author(s):  
Anikitos Garofalakis ◽  
Giannis Zacharakis ◽  
Heiko Meyer ◽  
Eleftherios N. Economou ◽  
Clio Mamalaki ◽  
...  

Author(s):  
Joanne Woodhouse ◽  
Alice Henley ◽  
Ross Lewin ◽  
John M Ward ◽  
Helen Hailes ◽  
...  

Green fluorescent protein (GFP), together with its family of variants, is the most widely used fluorescent protein for in vivo imaging. Numerous spectroscopic studies of the isolated GFP chromophore have...


1999 ◽  
Vol 339 (2) ◽  
pp. 299-307 ◽  
Author(s):  
Arthur L. KRUCKEBERG ◽  
Ling YE ◽  
Jan A. BERDEN ◽  
Karel van DAM

The Hxt2 glucose transport protein of Saccharomyces cerevisiae was genetically fused at its C-terminus with the green fluorescent protein (GFP). The Hxt2-GFP fusion protein is a functional hexose transporter: it restored growth on glucose to a strain bearing null mutations in the hexose transporter genes GAL2 and HXT1 to HXT7. Furthermore, its glucose transport activity in this null strain was not markedly different from that of the wild-type Hxt2 protein. We calculated from the fluorescence level and transport kinetics that induced cells had 1.4×105 Hxt2-GFP molecules per cell, and that the catalytic-centre activity of the Hxt2-GFP molecule in vivo is 53 s-1 at 30 °C. Expression of Hxt2-GFP was induced by growth at low concentrations of glucose. Under inducing conditions the Hxt2-GFP fluorescence was localized to the plasma membrane. In a strain impaired in the fusion of secretory vesicles with the plasma membrane, the fluorescence accumulated in the cytoplasm. When induced cells were treated with high concentrations of glucose, the fluorescence was redistributed to the vacuole within 4 h. When endocytosis was genetically blocked, the fluorescence remained in the plasma membrane after treatment with high concentrations of glucose.


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