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菲啶对酵母朊病毒[PSI+]的治愈效果的定量分析

[Quantification of the curing effects of phenanthridine on yeast prion [PSI+]].

作者信息

Zhong Zhengwei, Wang Lijie, Xie Hui, Li Hui, He Jianwei, Song Youtao

机构信息

School of Life Science, Liaoning University, Shenyang 110036, Liaoning, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2012 Jun;28(6):737-46.

Abstract

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.

摘要

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。

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