生物医学工程研究Issue(2)：93-97,5.
靶向大鼠TGF-β1基因shRN A真核表达载体的构建及鉴定
Construction and Identification of Eukaryotic Expression Plasmids Containing Short Hairpin RNA Targeting at Rat TGF-β1 gene
摘要
Abstract
To construct and identify eukaryotic expression plasmids containing short hairpin RNA(shRNA)that target at rat TGF-β1 gene and to provide a powerful tool for exploring a promising gene therapy way of hepatic fibrosis.Two pairs of shRNAs that target at rat TGF-β1 gene were designed,and no-isogeny gene segment HK was selected as a negative control.After annealed,the inserts were separately ligated into the linearized pGenesil-1 .1 plasmids.The eukaryotic expression plasmids pGenesil-TGF-β1-A,pGenesil-TGF-β1-B and pGenesil-HK were constructed,and then identified using restriction enzyme identification and sequencing analysis. Three recombinant plasmids were transfected into Hela cells according to Lipofectamine,and EGFP expressions were observed under fluorescence microscope after 48 h.By restriction enzyme identification and sequencing ,the DNA fragments were inserted corrranectly in pGenesil-1 .1 vectors as designed,respectively.Three recombinant plasmids were transfected into Hela cells.We successfully con-struct two eukaryotic expression recombinant plasmids pGenesil-1 .1 -shRNA targeted to rat TGF-β1 gene,and it will lay the foun-dation for further research on gene treatment of liver fibrosis.关键词
肝纤维化/TGF-β1/RN A干扰/质粒/基因重组Key words
Hepatic fibrosis/TGF-β1/RNA interference/Plasmid/Gene recombiant分类
医药卫生引用本文复制引用
都泓莲,张千,邓存良..靶向大鼠TGF-β1基因shRN A真核表达载体的构建及鉴定[J].生物医学工程研究,2014,(2):93-97,5.基金项目
四川省卫生厅课题（110345）。 （）
