Establishment of Transgenic cell model based on anti-oxidative response element / 生物医学工程学杂志
Journal of Biomedical Engineering
;
(6): 631-635, 2010.
Artigo
em Chinês
| WPRIM
| ID: wpr-230815
ABSTRACT
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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Timidina Quinase
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Transfecção
/
Elementos Facilitadores Genéticos
/
Transgenes
/
Elementos de Resposta
/
Proteínas de Fluorescência Verde
/
Células Hep G2
/
Vetores Genéticos
Limite:
Humanos
Idioma:
Chinês
Revista:
Journal of Biomedical Engineering
Ano de publicação:
2010
Tipo de documento:
Artigo
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