Construction and Identification of Eukaryotic Expression Plasmids Containing Short Hairpin RNA Targeting at Rat TGF-β1 gene
DU Honglian
ZHANG qian
DENG Cunliang
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.
Keywords：Hepatic fibrosisTGF-β1RNA interferencePlasmidGene recombiant
Publication Date：2014-01-01
Online Publishing Date：2026-09-11(First online date of this platform, not the publication date of the document)
Pages：5( 93-97 )