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p53 promoter-based reporter gene in vitro assays for quick assessment of agents with genotoxic potential
Authors:Li Huaixing  Shi Ke  Chen Ruiwen  He Yan  Wu Dan  Sun Shuhan
Affiliation:Department of Medical Genetics, Division of Basic Medical Sciences, Second Military Medical University, Shanghai, China.
Abstract:The p53 promoter-based green fluorescent protein(GFP)and luciferase reporter gene assays have been established for detecting DNA damage induced by genotoxic agents.To evaluate the system, NIH3T3 cells transfected with either pHP53-GFP or pMP53-GFP construct were treated with mitomycin or 5-fluorouracil.Expression of the GFP reporter gene was significantly and specifically induced in the cells exposed to mitomycin or 5-fluorouracil.Then we treated NIH3T3 cells harboring pHP53-Luc or pMP53- Luc vector with mitomycin,5-fluorouracil or cisplatin at various concentrations.Similarly,exposure of the cells to these agents with genotoxic potentials resulted in a dose-dependent induction in luciferase reporter gene expression.Thus,these in vitro reporter gene assays could provide an ideal system for quick assess- ment or screening of agents with genotoxic potential.
Keywords:p53 promoter  luciferase  green fluorescence protein(GFP)  genotoxic agent
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