Construction and identification of Sox9 siRNA expression vector

Objective To construct Sox9 siRNA expression vector in order to make clear the role of Sox9 in the genesis of chondrosarcoma.Methods According to the sequence of Sox9 mRNA demonstrated by Kou I and Ikegawa S,a pair of 64-nt oligonucleotides,each contain the sites of restriction endonuclease at both ends,were designed and synthesized The oligonucleotides were annealed and ligated with linearized pSilencer by T4DNA ligase.The combinants(designated ad pSilencer/Sox9 siRNA)were finally sequenced and identified by restriction endonuclease digestion.ResultsSox9 siRNA expression vector was successfully constructed and identified by double endonuclease digestion.Sequence analysis of the inserted fragment revealed the same sequence as that of the synthesized siRNA oligonucleotides.ConclusionSox9 siRNA expression vector has been successfully constructed,which paves the way for studying the role of Sox9 in chondrosarcoma,as well as in tumor therapy.