利用单分子追踪技术研究酿酒酵母活细胞中蛋白质动力学和靶标搜索机制。
Single-molecule tracking for studying protein dynamics and target-search mechanism in live cells of S. cerevisiae.
机构信息
Laboratory of Chromosome Dynamics and Gene Regulation, Department of Biotechnology, Indian Institute of Technology Hyderabad, Kandi, Sangareddy, Telangana 502285, India.
Laboratory of Chromosome Dynamics and Gene Regulation, Department of Biotechnology, Indian Institute of Technology Hyderabad, Kandi, Sangareddy, Telangana 502285, India.
出版信息
STAR Protoc. 2022 Dec 16;3(4):101900. doi: 10.1016/j.xpro.2022.101900. Epub 2022 Dec 5.
Single-molecule tracking (SMT) is a powerful approach to quantify the biophysical parameters of protein dynamics in live cells. Here, we describe a protocol for SMT in live cells of the budding yeast Saccharomyces cerevisiae. We detail how to genetically engineer yeast strains for SMT, how to set up image acquisition parameters, and how different software programs can be used to quantify a variety of biophysical parameters such as diffusion coefficient, residence time, bound fraction, jump angles, and target-search parameters. For complete details on the use and execution of this protocol, please refer to Mehta et al. and Ball et al...
单分子追踪（SMT）是一种强大的方法，可以定量测量活细胞中蛋白质动力学的生物物理参数。在这里，我们描述了在芽殖酵母酿酒酵母活细胞中进行 SMT 的方案。我们详细介绍了如何进行 SMT 的基因工程酵母菌株的构建，如何设置图像采集参数，以及如何使用不同的软件程序来定量测量各种生物物理参数，如扩散系数、停留时间、结合分数、跳跃角度和目标搜索参数。有关此方案的使用和执行的完整详细信息，请参考 Mehta 等人和 Ball 等人的文献。
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