Abstract: In present research,we intend to construct pG-super-siPrP expression plasmid and explore its function in C6 cells．shPrP was subcloned into pG-super. Recombinant plasmid pG-super-shPrP was transformed into Top10 E.coli, and the ampicillin resistant clones were identified by PCR and DNA sequencing．C6 cells were transfected with the identified pG-super-shPrP and pG-super (the control group) by LipofectamineTM2000．PrP mRNA was quantified by realtime RT-PCR.The results showed that the expression of PrP mRNA in pG-super-shPrP group decreased by 34.2% compared with the control group (P<0.05)．The detection results of the total activity of superoxide dismutase indicated that PrP promoted the SOD activity.In conclusion, pG-super-shPrP expression plasmid was constructed effectively and took effect in C6 cells，which helped to further study about PrP biochemical and physiological role in animal and offer a attractive therapeutic approach to fight against prion disease.