Stathmin SiRNA Plasmid Expression Vector Suppressing Expression of Stathmin

Objective:Constructing stathmin specific SiRNA plasmid expression vector and investigating the silence effect of SiRNA plasmid expression vector on stathmin in 5-8F cell line.Method:Synthesize stathmin specific interfering expression DNA fragments,obtain doubled DNA fragment by annealing and combine the fragment with plasmid pGenesil1.1.The recombinant vectors were transferred and screened in JM109 strains.Then,the recombinant vectors were amplified and identified by di-enzyme digestion and DNA sequencing.The identified vector was transfected into 5-8F cell line.RT-PCR and Western Blot were adopted to analyze the expression of stathmin.Result:Di-enzyme digestion analysis and DNA sequencing showed that the basic sequence and direction of stathmin specific DNA fragment,which was inserted into SiRNA plasmid expression,was correct and reduced the expression of stathmin effectively in nasopharyngeal carcinoma cell line 5-8F.The transfected efficiency was about 78.8±6.8%.Conclusion:The constructed stathmin specific recombinant SiRNA plasmid expression vector could silence the expression of stathmin.