Inhibition of survivin gene expression in EJ cells by RNAi / 重庆医科大学学报

ABSTRACT

Objective:To construct eukaryotic expression vectors with the pSilencer2.0 vector for inhibiting human survivin gene by RNA interference,and to detect the effect of the silenced survivin gene on EJ cells.Methods:Two target gene segments were synthesized and cloned into the pSilencer2.0 vector respectively to construct two recombinant eukaryotic expression vectors,pSilencer2.0-SVV1and pSilencer2.0-SVV2,which were identified by enzyme digestion analysis and DNA sequencing.Then the EJ cells were transfected with the recombinant vectors by lipofection and the interference effect was detected by RT-PCR and Western-blot.Results:Enzyme digestion analysis and DNA sequencing showed that two target segments were cloned into pSilencer2.0 vectors correctly.The results of RT-PCR and Western blot indicated that pSilencer2.0-SVV1 and pSilencer2.0-SVV2 vectors could significantly knock down the transcription and expression of survivin gene(P