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Chinese Journal of Hepatology ; (12): 196-198, 2007.
Artigo em Chinês | WPRIM | ID: wpr-285431

RESUMO

<p><b>OBJECTIVE</b>To construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells.</p><p><b>METHODS</b>The combinant lone DNA with cutting sites of BamH I and Xho I enzyme according to the sequences of 447-465, 552-540 TIMP-1 of rats and nonspecific sequence were selected and cloned to pGEM-T vector and sub-cloned to pRNAT-U6.2. They were then identified by double enzyme digestion analysis and DNA sequencing. Three plasmids were transfected into T6 separately through an oligofectamine package. TIMP-1 mRNA expression was evaluated by RT-PCR.</p><p><b>RESULTS</b>Targeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6.</p><p><b>CONCLUSION</b>We successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression.</p>


Assuntos
Animais , Ratos , Linhagem Celular , Inativação Gênica , Vetores Genéticos , Células Estreladas do Fígado , Plasmídeos , RNA Interferente Pequeno , Inibidor Tecidual de Metaloproteinase-1 , Genética , Transfecção
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