Construction and identification of eukaryotic expression vector of short hairpin RNA for IER3IP1 gene
- VernacularTitle:针对IER3IP1基因的shRNA真核表达载体的构建和鉴定
- Author:
Yan LEI
- Publication Type:Journal Article
- Keywords:
RNA interference;
Gene,IER3IP1
- From:
Journal of Chongqing Medical University
2003;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Toconstruct the shRNA(Short Hairpin RNA)eukaryotic expression vector targetingat IER3IP1 gene and observe its silencing effect on IER3IP1 gene.Methods:Two pairs of DNA fragments targeting at IER3IP1 gene and one pair of DNA fragment of negative control were synthesized and cloned into the Eukaryotic Expression Vector pGenesil-1.These vectors were separately transfected into the L02 cells.Inhibitory effect at the time of 24,48,72 h after transfection were detected by semiquantitative RT-PCR.Results:These shRNA eukaryotic expression vectors were successfully constructed.One of them could significantly and specially inhibite the expression of IER3IP1 gene and the inhibition ratios were separately 64%,79%and 69%at the time of 24,48 and 72 h,compared with empty vector group.Conclusion:The combinated eukaryotic expression vector could inhibit the expression of IER3IP1 gene,which forms the basis for investigating IER3IP1 gene's function.
