Monitoring of Electrophile Chemical Pollutants Using Transgenic Firefly Luciferase Gene Cells / 环境与健康杂志

ABSTRACT

Objective To assess quickly the potential environmental pollution by using the luciferase transgenic cells. Methods Many electrophile compounds in the environment can generate oxidative stress,so that the transcription of certain protective genes is induced via specific DNA motifs called electrophile response elements (EPREs). We have made a vector containing a single EPRE fused to the TK minimal promoter and the gene encoding firefly luciferase (EPRE-LUC) by adopting bio-molecular techniques. From this vector the stable LUC expression vector regulated by EPRE had been successfully reconstructed. This reporter construct was transfected into HeLa cells,and the clones resistant to G418 were selected. The resistant cells were treated by the different concentrations of sodium arsenate(NaAsO2),cadmium chloride (CdCl2),mercury chloride(HgCl2) and diethyl mateate (DEM). After that,the expression of luciferase was determined by luciferase assay kit. Results The correct construct frame of LUC reporter vector regulated by EPRE was identified by DNA sequencing; the dose-dependent relationships between the LUC expression and the test substance concentration were found. Among them,the relationship produced by DEM was the most significant. Conclusion The LUC transgenic cells regulated by EPRE have been successfully constructed.