Quantitative determination of protein stability and ligand binding using a green fluorescent protein reporter system

2010 ◽  
Vol 6 (7) ◽  
pp. 1285 ◽  
Author(s):  
Morgane J. J. Moreau ◽  
Isabelle Morin ◽  
Patrick M. Schaeffer
Keyword(s):  
Green Fluorescent Protein ◽  
Ligand Binding ◽  
Quantitative Determination ◽  
Protein Stability ◽  
Fluorescent Protein ◽  
Green Fluorescent Protein Reporter ◽  
Reporter System ◽  
Green Fluorescent ◽  
Fluorescent Protein Reporter

GATA-1 expression pattern can be recapitulated in living transgenic zebrafish using GFP reporter gene

Development ◽  
1997 ◽  
Vol 124 (20) ◽  
pp. 4105-4111 ◽  
Author(s):  
Q. Long ◽  
A. Meng ◽  
H. Wang ◽  
J.R. Jessen ◽  
M.J. Farrell ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Progenitor Cells ◽  
Reporter Gene ◽  
Fluorescent Protein ◽  
Expression Patterns ◽  
Transgenic Fish ◽  
Green Fluorescent Protein Reporter ◽  
Erythroid Progenitor ◽  
Green Fluorescent ◽  
Fluorescent Protein Reporter

In this study, DNA constructs containing the putative zebrafish promoter sequences of GATA-1, an erythroid-specific transcription factor, and the green fluorescent protein reporter gene, were microinjected into single-cell zebrafish embryos. Erythroid-specific activity of the GATA-1 promoter was observed in living embryos during early development. Fluorescent circulating blood cells were detected in microinjected embryos 24 hours after fertilization and were still present in 2-month-old fish. Germline transgenic fish obtained from the injected founders continued to express green fluorescent protein in erythroid cells in the F1 and F2 generations. The green fluorescent protein expression patterns in transgenic fish were consistent with the pattern of GATA-1 mRNA expression detected by RNA in situ hybridization. These transgenic fish have allowed us to isolate, by fluorescence-activated cell sorting, the earliest erythroid progenitor cells from developing embryos for in vitro studies. By generating transgenic fish using constructs containing other zebrafish promoters and green fluorescent protein reporter gene, it should be possible to visualize the origin and migration of any lineage-specific progenitor cells in a living embryo.

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