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.