A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress

Molina, A.; Carpeaux, Rudy; Martial, Joseph; Muller, Marc

doi:10.1016/S0887-2333(01)00106-0

Article (Scientific journals)

A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress

Molina, A.; Carpeaux, Rudy; Martial, Joseph et al.

2002 • In Toxicology in Vitro, 16 (2), p. 201-7

Peer Reviewed verified by ORBi

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https://hdl.handle.net/2268/13023

DOI
10.1016/S0887-2333(01)00106-0

PubMed
11869883

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Keywords :

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

Abstract :

[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.

Research Center/Unit :

AFFISH-RC - Applied and Fundamental FISH Research Center - ULiège
Giga-Development and Stem Cells - ULiège

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Molina, A.

Carpeaux, Rudy ; Université de Liège - ULiège > Département des sciences cliniques > GIGA-R : Hématologie

Martial, Joseph ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire

Muller, Marc ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire

Language :

English

Title :

A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress

Publication date :

2002

Journal title :

Toxicology in Vitro

ISSN :

0887-2333

eISSN :

1879-3177

Publisher :

Pergamon-Elsevier Science Ltd, Oxford, United Kingdom

Volume :

Issue :

Pages :

201-7

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 22 May 2009

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