定量研究菲啶对酵母朊病毒PSI+的治愈作用

摘要

为系统研究菲啶对酵母朊病毒的治愈效果,借助表达融合蛋白 GFP-Sup35p 的酵母朊病毒模型(NGMC),引入半变性琼脂糖凝胶电泳技术和荧光漂白后恢复技术在蛋白和细胞水平定量分析了菲啶对酵母朊病毒的治愈效果.结果表明,蛋白和细胞水平采用的定量分析方法能够精确定量菲啶对酵母朊病毒的治愈作用,菲啶作用酵母朊病毒[PSI+] 1～5 d的治愈率分别为0％、0％、51.7％、87.5％和94.4％.另外,菲啶作用酵母朊病毒[PSI+]细胞1～2 d后出现的粉色菌落中朊病毒的聚集状态与[PSI+]相似,而3～5 d后出现的粉色菌落中朊病毒的状态与[psi-]相似.%In order to quantify the curing effects of phenanthridine on yeast prion, we introduced semi-denaturing agarose gel electrophoresis and fluorescence recovery after photobleaching techniques to quantify the curing effects of phenanthridine on yeast prion at the protein and cellular levels with the [PSI+] yeast strain expressing GFP-Sup35p (NGMC) . The results showed that these two approaches could precisely quantify the curing effects of phenanthridine on [PSI+] cells. After a treatment for 1 through 5 days with phenanthridine, the curing rates of [PSI+] cells were 0%, 0%, 51.7%, 87.5% and 94.4%, respectively. Meanwhile, we quantified the sizes of Sup35p polymers in phenanthridine induced pink phenotype cells. The aggregation status in 1-2 days phenanthridine treated cells were similar to those in [PSI+] cells, while the aggregation status in 3-5 days phenanthridine treated cells were similar to those in [psi-] cells.