Construction and Identification of Rab25 siRNA Expression Vector

Objective To construct Rab25 siRNA expression vector in order to explore the new method of ovarian cancer gene therapy.Methods According to Rab25 mRNA sequence in the Genebank,a pair of 64-nt oligonucleotides,each containing the sites of restriction endonuclease at both ends,were designed and synthesized.Oligonucleotides were annealed and ligated with linearized pSUPER by T4DNA ligase.The recombinants(named pSUPER /Rab25 siRNA) were finally sequenced and identified by enzyme cutting and sequencing.Results Rab25 siRNA expression vector was successfully constructed and identified by double endonuclease digestion.Sequence analysis of inserted fragment revealed the same sequence as synthesized siRNA oligonucleotides.Conclusion Rab25 siRNA expression vector has been successfully constructed,which will facilitate further studies of Rab25 function and its application in the treatment of ovarian cancer.