Green Fluorescent Protein as a Real Time Quantitative Reporter of Heterologous Protein Production

1998 ◽  
Vol 14 (2) ◽  
pp. 351-354 ◽  
Author(s):  
C.R. Albano ◽  
L. Randers-Eichhorn ◽  
W.E. Bentley ◽  
G. Rao
Keyword(s):  
Green Fluorescent Protein ◽  
Real Time ◽  
Protein Production ◽  
Heterologous Protein ◽  
Fluorescent Protein ◽  
Heterologous Protein Production ◽  
Green Fluorescent

scholarly journals Novel Real-Time Monitoring System for Human Cytomegalovirus- Infected Cells In Vitro That Uses a Green Fluorescent Protein-PML-Expressing Cell Line

2006 ◽  
Vol 50 (8) ◽  
pp. 2806-2813 ◽  
Author(s):  
T. Ueno ◽  
Y. Eizuru ◽  
H. Katano ◽  
T. Kurata ◽  
T. Sata ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Cell Line ◽  
Real Time ◽  
Human Cytomegalovirus ◽  
Hcmv Infection ◽  
Fluorescent Protein ◽  
Clinical Isolates ◽  
Pml Bodies ◽  
Infected Cells ◽  
Green Fluorescent

ABSTRACT Promyelocytic leukemia (PML) bodies are discrete nuclear foci that are intimately associated with many DNA viruses. In human cytomegalovirus (HCMV) infection, the IE1 (for “immediate-early 1”) protein has a marked effect on PML bodies via de-SUMOylation of PML protein. Here, we report a novel real-time monitoring system for HCMV-infected cells using a newly established cell line (SE/15) that stably expresses green fluorescent protein (GFP)-PML protein. In SE/15 cells, HCMV infection causes specific and efficient dispersion of GFP-PML bodies in an IE1-dependent manner, allowing the infected cells to be monitored by fluorescence microscopy without immunostaining. Since a specific change in the detergent solubility of GFP-PML occurs upon infection, the infected cells can be quantified by GFP fluorescence measurement after extraction. With this assay, the inhibitory effects of heparin and neutralizing antibodies were determined in small-scale cultures, indicating its usefulness for screening inhibitory reagents for laboratory virus strains. Furthermore, we established a sensitive imaging assay by counting the number of nuclei containing dispersed GFP-PML, which is applicable for titration of slow-growing clinical isolates. In all strains tested, the virus titers estimated by the GFP-PML imaging assay were well correlated with the plaque-forming cell numbers determined in human embryonic lung cells. Coculture of SE/15 cells and HCMV-infected fibroblasts permitted a rapid and reliable method for estimating the 50% inhibitory concentration values of drugs for clinical isolates in susceptibility testing. Taken together, these results demonstrate the development of a rapid, sensitive, quantitative, and specific detection system for HCMV-infected cells involving a simple procedure that can be used for titration of low-titer clinical isolates.

Rapid Optimization of Membrane Protein Production Using Green Fluorescent Protein-Fusions and Lemo21(DE3)

2011 ◽  
pp. 391-406
Author(s):  
Susan Schlegel ◽  
Mirjam Klepsch ◽  
Dimitra Gialama ◽  
David Wickström ◽  
David Drew ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Membrane Protein ◽  
Protein Production ◽  
Fluorescent Protein ◽  
Green Fluorescent

scholarly journals Real-Time Optical Monitoring of Ligand-Mediated Internalization of α1b-Adrenoceptor with Green Fluorescent Protein

1998 ◽  
Vol 12 (8) ◽  
pp. 1099-1111 ◽  
Author(s):  
Takeo Awaji ◽  
Akira Hirasawa ◽  
Masakazu Kataoka ◽  
Hitomi Shinoura ◽  
Yasuhisa Nakayama ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Real Time ◽  
Fluorescent Protein ◽  
Optical Monitoring ◽  
Green Fluorescent

scholarly journals Real-time light-driven dynamics of the fluorescence emission in single green fluorescent protein molecules

2000 ◽  
Vol 97 (13) ◽  
pp. 7237-7242 ◽  
Author(s):  
M. F. Garcia-Parajo ◽  
G. M. J. Segers-Nolten ◽  
J.-A. Veerman ◽  
J. Greve ◽  
N. F. van Hulst
Keyword(s):  
Green Fluorescent Protein ◽  
Real Time ◽  
Fluorescent Protein ◽  
Fluorescence Emission ◽  
Protein Molecules ◽  
Green Fluorescent

Rapid and massive green fluorescent protein production leads to formation of protein Y-bodies in plant cells

2012 ◽  
Vol 77 (6) ◽  
pp. 603-608 ◽  
Author(s):  
T. V. Komarova ◽  
E. V. Sheval ◽  
D. V. Pozdyshev ◽  
V. S. Kolesnikova ◽  
Yu. L. Dorokhov
Keyword(s):  
Green Fluorescent Protein ◽  
Protein Production ◽  
Fluorescent Protein ◽  
Plant Cells ◽  
Green Fluorescent Protein Production ◽  
Green Fluorescent
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