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

RESUMO

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.


Assuntos
Humanos , Antineoplásicos , Farmacologia , Sequência de Bases , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Métodos , Elementos Facilitadores Genéticos , Genética , Expressão Gênica , Gentamicinas , Farmacologia , Proteínas de Fluorescência Verde , Genética , Metabolismo , Células Hep G2 , Lentinano , Farmacologia , Microscopia de Fluorescência , Dados de Sequência Molecular , Oligonucleotídeos , Genética , Prolina , Farmacologia , Proteínas Recombinantes de Fusão , Genética , Metabolismo , Tiocarbamatos , Farmacologia , Transfecção
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