中文摘要：

本文旨在构建有效抑制朊蛋白（Prion protein，PrP）表达的重组质粒，并以此为工具控制PrP表达，从而探讨PrP对细胞SOD活性的影响。设计并化学合成1对含有发夹结构的寡核苷酸片段（shPrP），退火后与表达载体pG-super（Hairpin siRNA expressing vector）定向连接，构建重组质粒pG-super-shPrP。对重组子进行PCR鉴定，测序正确后，脂质体法转染C6细胞，采用实时荧光定量RT-PCR检测PrP mRNA的表达水平，以验证pG-super-shPrP的抑制效率；结果表明：重组质粒pG-super-shPrP构建成功，且显著降低C6细胞PrP mRNA表达（P〈0．05），抑制效率为34．2％。利用pG-super、pG-super-shPrP分别转染C6细胞，并检测细胞SOD总活性及SOD表达水平，探讨PrP对细胞SOD活性的影响及其作用机制，结果表明PrP促进细胞SOD的活性（P〈0．01），但对细胞SOD的表达量无影响，即PrP对SOD活性的促进作用与SOD1的表达量无关。本研究在成功构建了PrP的RNA干扰表达质粒的基础上，利用此质粒，在细胞水平上揭示了PrP对细胞SOD活性的促进作用。

英文摘要：

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 Topl0 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 LipofectamineTM 2000. 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.