中文摘要：

参考Saborio和Soto创建的名为蛋白质错误折叠循环扩增（protein misfolding cyclic amplification,PMCA）的方法建立一种22L毒株系朊蛋白错误折叠循环扩增方法,以便研究22L毒株系异常朊蛋白（scrapie PrP,PrP^Sc）的合成机制。在阴离子去污剂十二烷基硫酸钠（SDS）存在的条件下,这种方法可以通过超声降解从而重复扩增产物,并在仓鼠脑匀浆中快速有效的模仿体内朊病毒PrP^Sc的复制,因此可以在几个小时内高浓度扩增PrP^Sc。已有试验结果发现,PrP^Sc体外转化率在使用粗提的脑组织匀浆时比使用纯化重组PrP更高,这个发现暗示着高效率的PrP^Sc转化可能还需要探索其他未知的宿主因素。研究结果表明,建立的这种改良的PMCA技术可用于研究PrP^Sc合成的生物化学机制,以此来进行诊断与治疗方法的研究。

英文摘要：

Establishing a method of PMCA for 22L strain PrP^Sc based on the method called protein misfolding cyclic amplification（PMCA） established by Saborio and Soto to research the biochemical mechanism of PrP^Sc formation.Under the existence of the anion detergent lauryl sodium sulfate（SDS）,the method can directly degrade the products that producted by repeated amplification,and can in the brain tissue homogenate of hamster effectively imitate the replication mode that PrP^Sc run in vivo in a accelerated way.Therefore,it can amplify PrP^Sc to high concentrations in a few hours.We found that in vitro unbolted brain tissue homogenate is more efficiently than purified reconstituted about PrP^Sc conversion,this discovery implied that highly efficient PrP^Sc conversion may associated with other unknown host factor.Researching indicated that the improved PMCA technique that we established can be used to research the biochemical mechanism of PrP^Sc formation.