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Establishment of a transgenic cell model for preliminary screening of chemopreventive agents / 生物工程学报
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)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Oligonucleotides / Pharmacology / Thiocarbamates / Recombinant Fusion Proteins / Proline / Drug Screening Assays, Antitumor / Molecular Sequence Data / Base Sequence / Gentamicins / Transfection Type of study: Diagnostic study / Prognostic study / Screening study Limits: Humans Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2007 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Oligonucleotides / Pharmacology / Thiocarbamates / Recombinant Fusion Proteins / Proline / Drug Screening Assays, Antitumor / Molecular Sequence Data / Base Sequence / Gentamicins / Transfection Type of study: Diagnostic study / Prognostic study / Screening study Limits: Humans Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2007 Type: Article