Reference : A transformed fish cell line expressing a green fluorescent protein-luciferase fusion ge...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/13023
A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress
English
Molina, A. [> > > >]
Carpeaux, Rudy [Université de Liège - ULg > Département des sciences cliniques > GIGA-R : Hématologie >]
Martial, Joseph mailto [Université de Liège - ULg > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire >]
Muller, Marc mailto [Université de Liège - ULg > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire >]
2002
Toxicology in Vitro
Pergamon-Elsevier Science Ltd
16
2
201-7
Yes (verified by ORBi)
International
0887-2333
Oxford
United Kingdom
[en] Animals ; Cell Line, Transformed/drug effects/*metabolism ; Dose-Response Relationship, Drug ; Flow Cytometry ; Genes, Reporter ; Green Fluorescent Proteins ; HSP70 Heat-Shock Proteins/genetics/metabolism ; Indicators and Reagents/*metabolism ; Luciferases/genetics/*metabolism ; Luminescent Proteins/genetics/*metabolism ; Recombinant Fusion Proteins/genetics/*metabolism ; Tilapia ; Xenobiotics/toxicity
[en] We obtained a stable transformed fish (EPC) cell line containing a reporter gene under the control of the tilapia HSP70 promoter. Expression of the reporter gene, coding for a green fluorescent protein (GFP)-luciferase fusion protein, was assessed by measuring the luciferase enzymatic activity by luminometry and the GFP expression by fluorescence microscopy and flow cytometry. The clone was characterized for its capacity to respond to heat shock treatment. The results show high induction after 1 h at 37 degrees C of treatment, up to 500-fold. In addition, its convenience to detect a large range of cellular stressors was evaluated. We observed high induction when Cd2+, Zn2+, Hg2+ or Cu2+ was added, but not Pb2+. In addition, activation of the reporter gene was observed in the presence of other compounds such as acetyl chloride, tetrachlorophenol, chloroacetamide and sodium arsenite. In conclusion, this cell line can be used as a rapid, cheap and easy biological test to determine cellular stress induced by environmental pollutants, alone or in conjunction with other, more specific assays.
Applied and Fundamental FISH Research Center - AFFISH-RC ; Giga-Development and Stem Cells
http://hdl.handle.net/2268/13023
10.1016/S0887-2333(01)00106-0
2002/03/01

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