Construction and Identification of shRNA Expression Vector for Human Rac1 Gene

Objective To construct the shRNA expression vector for human Rac1 gene and lay a foundation of increasing the sensitivity of ovarian cancer to radiotherapy and chemotherapy.Methods The oligo DNA for construction of shRNA expression vector was designed and synthesized by using shRNA design software according to the sequence of human Rac1 gene in GenBank,based on which recombinant shRNA vector with human Rac1 gene was constructed and tranfected to ovarian cancer Skov3 cells,and effective recombinant shRNA plasmid was screened by RT-PCR 48 h later.Results Recombinant shRNA plasmid with Rac1 gene was identified by digestion with BamHⅠ and PstⅠ,of which the sequence was completely consistent with theoretical sequence.RT-PCR showed that the transcription level of Rac1 mRNA in Skov3 cells transfected with plasmid pGPU6/GFP/Rac1-524 decreased.Conclusion The shRNA expression vector for human Rac1 gene was successfully constructed,and effective interfering plasmid pGFU6/GFP/Rac1-524 was screened.