Construction and identification of the RNA interference system targeting CGI-100 gene
- VernacularTitle:靶向CGI-100基因shRNA干扰载体的构建与鉴定
- Author:
Yongqiang WANG
- Publication Type:Journal Article
- Keywords:
CGI-100 gene;
RNA interference;
shRNA;
K562 cell
- From:
Journal of Chongqing Medical University
2007;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the expression vector of RNA interference(RNAi)system for CGI-100 gene and identify its inhibition efficiency.Methods:Three oligonucleotides targeting CGI-100 gene were devised and chemically synthesized,annealed to form double-strand DNA and inserted into plasmid PGenesil-1 whose segments were digested by BamHⅠ and HindⅢ.The recombinant vector were then transfected into K562 cells with lipofectamineTM 2000.CGI-100 expresstion in the transfected cells was assayed by RT-PCR and dot blot.Results:The recombinant plasmids of PGenesil-1-CGI-100 were successfully constructed by DNA sequencing analysis.The first of PGenesil-1-CGI-100 had the highest inhibition efficiency in mRNA levels,up to 54%.Conclusion:The RNA interference system targeting CGI-100 gene were successfully constructed and a interference vector with a higher inhibition efficiency was screened,which provides the foundation for further investigation.
