使用光漂白后荧光恢复技术监测 中的冷凝物动态。
Monitoring condensate dynamics in using fluorescence recovery after photobleaching.
机构信息
Department of Chemistry, Washington University, St. Louis, MO 63130, USA.
Department of Chemistry, Washington University, St. Louis, MO 63130, USA.
出版信息
STAR Protoc. 2022 Jul 31;3(3):101592. doi: 10.1016/j.xpro.2022.101592. eCollection 2022 Sep 16.
This protocol describes the use of fluorescence recovery after photobleaching (FRAP) to investigate the dynamics of Matrin-3 (MATR3) condensates in live budding yeast. We detail how to generate yeast strains containing MATR3 with an enhanced green fluorescent protein (eGFP) tag and induce MATR3-eGFP expression. We provide steps to prepare slides of immobilized yeast cells and perform FRAP imaging and data analysis. This protocol can be broadly applied to study condensate dynamics of a range of proteins in different model systems. For complete details on the use and execution of this protocol, please refer to Sprunger et al. (2022).
本方案描述了使用光漂白后荧光恢复（FRAP）技术来研究活 budding 酵母中 Matrin-3（MATR3）凝聚物的动力学。我们详细介绍了如何生成含有增强型绿色荧光蛋白（eGFP）标签的 MATR3 的酵母菌株，并诱导 MATR3-eGFP 表达。我们提供了制备固定化酵母细胞载玻片以及进行 FRAP 成像和数据分析的步骤。本方案可广泛应用于研究不同模型系统中一系列蛋白质凝聚物的动力学。有关该方案使用和执行的完整详细信息，请参阅 Sprunger 等人（2022 年）。
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