title: [Construction and identification of matrix metalloproteinase inhibitor-1 siRNA eukaryotic expression vectors].
abstract: OBJECTIVE
  To construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells.
  
  
  METHODS
  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.
  
  
  RESULTS
  Targeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6.
  
  
  CONCLUSIONS
  We successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression.
authors.affiliation: Department of Gastroenterology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. tianyu615999@sina.com
authors.forename: Li
authors.initials: L
authors.lastname: Tian
authors.forename: Ji-chang
authors.initials: JC
authors.lastname: Li
authors.forename: Guo-qiang
authors.initials: GQ
authors.lastname: Zhao
authors.forename: Kui-sheng
authors.initials: KS
authors.lastname: Chen
authors.forename: Wen-hui
authors.initials: WH
authors.lastname: Fan
authors.forename: Xin
authors.initials: X
authors.lastname: Lou
authors.forename: Miao-miao
authors.initials: MM
authors.lastname: Sun
authors.forename: Xue-quan
authors.initials: XQ
authors.lastname: Cao
mesh_terms: D000818:Animals; D002460:Cell Line; D020868:Gene Silencing; D005822:Genetic Vectors; D055166:Hepatic Stellate Cells; D010957:Plasmids; D034741:RNA, Small Interfering; D051381:Rats; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics*; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering; D019715:Tissue Inhibitor of Metalloproteinase-1