[Establishment of Transgenic cell model based on anti-oxidative response element].
To establish a transgenic cell model based on anti-oxidative response element (ARE) and green fluorescence protein(GFP) reporter gene, the TK minimal promoter was amplified by PCR and cloned into pEGFP-N1 for constructing reporter vector pTK-GFP/Neo. Four synthetic oligonucleotide ARE motifs were annealed and purified and then were inserted into pTK-GFP/Neo one by one to construct the eukaryotic reporter vector p4ARE-TK-GFP/Neo. Two reconstruct eukaryotic reporter vectors were transfected into HepG2 cells mediated by lipofectamine. The positive clones were obtained by the screen of G418. The cell model was tested with PDTC and tBHQ, well known inducers of phase II enzymes, by determining GFP activity. The results showed that the expression level of GFP was significantly increased by PDTC and tBHQ, and a transgenic cell model based on ARE was established successfully.
Xu
Hairong
H
Institute of Integrated Chinese and Western Medicine, Medical College of Yangzhou University, Yangzhou 225001, China.
Bo
Ping
P
Li
Xiangming
X
D000975:Antioxidants / Q000378:metabolism*; D001483:Base Sequence; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D056945:Hep G2 Cells; D006801:Humans; D008969:Molecular Sequence Data; D020218:Response Elements / Q000235:genetics*; D013937:Thymidine Kinase / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D019076:Transgenes
D000975:Antioxidants; D049452:Green Fluorescent Proteins; D013937:Thymidine Kinase