Construction of eukaryotic expression plasmid experssing shRNA targeting EGFR gene family
- VernacularTitle:RNA干扰EGFR基因家族真核表达载体的构建
- Author:
Huan HUANG
- Publication Type:Journal Article
- Keywords:
EFGR family;
RNA interference;
Eukarvotic expression vector
- From:
Journal of Chongqing Medical University
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the eukaryotic expression plasmid expressing shRNA targeting EGFR gene family as a tool for following experiments in vitro and in vivo.Methods:EFGR family highly expressed in low radiosensitive ovarian cancer cells SKOV3,chosen highly expressed homologous genes in EFGR family as targeted gene.According to guidelines for shRNA design,designed experimental shRNA and negative control shRNA.In vitro synthesized 2 of the coding sequence of short hairpin RNA DNA single-strands,annealing,and cloned into the corresponding vector pGensil-1 of pol Ⅲ promoter downstream,named pGensil-1-E and pGensil-1-C.Confirmed by restrict endonuclease digestion and DNA sequencing.Conclusions:Eukaryotic expression plasmid experssing shRNA targeting homologous genes in EFGR family was constructed successfully.
