Objective：To construct and identify a recombinant survivin-shRNA expression vector, for making a foundation of keloid gene therapy via RNA interference. Methods：According to survivin cDNA gene sequence in the gene bank, a pair of 65 nt oligonucleotides each containing the sites of restriction endonuclease at both ends were designed and synthesized by Reynolds design principles. Oligonucleotides were annealed and ligated with linedrized RNAi-Ready pSIREN-DNR-DsRed-Express by T4DNA ligase. The recombinants were transfected into Escherichia coli strains DH-5a. Finally, it was sequenced and identified by restriction endonuclease digestion. Results：The size of the target gene fragment amplified by PCR was 465 bp and in accordance with the expected results. Double digested section showed a target gene band of about 65 bp identitfied by double endonuclease digestion. Sequence analysis of inserted fragment revealed the same sequence as synthesized shRNA oligonucleotides. Conclusions：A survivin-shRNA expression vector has been successfully constructed, which can be as the groundwork for future study of liposome mediated transfection into scar or tumor cells.