Zhong Zhengwei, Wang Lijie, Xie Hui, Li Hui, He Jianwei, Song Youtao
School of Life Science, Liaoning University, Shenyang 110036, Liaoning, China.
Sheng Wu Gong Cheng Xue Bao. 2012 Jun;28(6):737-46.
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.
为了量化菲啶对酵母朊病毒的治愈效果,我们引入了半变性琼脂糖凝胶电泳和光漂白后荧光恢复技术,以在蛋白质和细胞水平上,用表达绿色荧光蛋白标记的 Sup35p(NGMC)的[PSI+]酵母菌株来量化菲啶对酵母朊病毒的治愈效果。结果表明,这两种方法能够精确量化菲啶对[PSI+]细胞的治愈效果。用菲啶处理1至5天后,[PSI+]细胞的治愈率分别为0%、0%、51.7%、87.5%和94.4%。同时,我们量化了菲啶诱导的粉红色表型细胞中Sup35p聚合物的大小。经菲啶处理1至2天的细胞中的聚集状态与[PSI+]细胞中的相似,而经菲啶处理3至5天的细胞中的聚集状态与[psi(-)]细胞中的相似。