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Chinese Journal of Biotechnology ; (12): 85-89, 2007.
Article in Chinese | WPRIM | ID: wpr-325414

ABSTRACT

To develop a GFP transgenic cell model under the transcriptional control of TK promoter adjacent to which ARE enhancer was inserted. Synthetic oligonucleotide ARE motif was annealed and purified then inserted into pTK-GFP to construct the vector of pARE-TK-GFP. The TK and ARE-TK fragments were amplified by PCR and cloned into pEGFP-N1 to reconstruct eukaryotic expression vectors of pTK-GFP/Neo and pARE-TK-GFP/Neo. They were transfected into HepG2 cells and clones resistant G418 were isolated. PDTC and Lentinan were used to induce the cell levels of GFP and the fluorescence was measured using a fluorescence plate reader. The results showed that the induced level of GFP is significantly increased and have dose-dependeny in a certain range. This findings indicated that such a cell model offered a potential platform for preliminary screening of all kinds of natural or synthetic chemopreventive agents.


Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Base Sequence , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Methods , Enhancer Elements, Genetic , Genetics , Gene Expression , Gentamicins , Pharmacology , Green Fluorescent Proteins , Genetics , Metabolism , Hep G2 Cells , Lentinan , Pharmacology , Microscopy, Fluorescence , Molecular Sequence Data , Oligonucleotides , Genetics , Proline , Pharmacology , Recombinant Fusion Proteins , Genetics , Metabolism , Thiocarbamates , Pharmacology , Transfection
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