Department of Genetics, Development, and Cell Biology, Iowa State University, Ames, IA 50010, USA.
Department of Genetics, Development, and Cell Biology, Iowa State University, Ames, IA 50010, USA.
STAR Protoc. 2023 Sep 15;4(3):102522. doi: 10.1016/j.xpro.2023.102522. Epub 2023 Aug 18.
The microtubule (MT)-stabilizing drug Taxol (paclitaxel) is a commonly used tool to investigate MT dynamics and MT-dependent processes. Here, we present a protocol for using Taxol-sensitized budding yeast to investigate the effect of microtubule stabilization on anaphase onset. We describe steps for establishing a log phase culture, synchronizing cells in G1, arresting in metaphase, and releasing cells into Taxol. We then detail procedures for imaging and scoring anaphase onset. This protocol facilitates maintenance and reproducibility in testing drug-sensitized and Taxol-sensitized yeast strains. For complete details on the use and execution of this protocol, please refer to Proudfoot et al..
微管（MT）稳定剂紫杉醇（紫杉醇）是一种常用于研究 MT 动力学和 MT 依赖过程的工具。在这里，我们提出了一个使用紫杉醇敏感的出芽酵母来研究微管稳定对后期起始影响的方案。我们描述了建立对数期培养物、将细胞同步在 G1 期、在中期阻断以及将细胞释放到紫杉醇中的步骤。然后，我们详细介绍了后期起始的成像和评分程序。该方案有助于在测试药物敏感和紫杉醇敏感酵母菌株时保持和重现性。有关此方案的使用和执行的完整详细信息，请参阅 Proudfoot 等人的研究。
