利用邻近连接分析原位检测秀丽隐杆线虫生殖系中的核糖核蛋白复合体组装
In Situ Detection of Ribonucleoprotein Complex Assembly in the C. elegans Germline using Proximity Ligation Assay.
作者信息
Day Nicholas J, Wang Xiaobo, Voronina Ekaterina
机构信息
Division of Biological Sciences, University of Montana.
Division of Biological Sciences, University of Montana;
出版信息
J Vis Exp. 2020 May 5(159). doi: 10.3791/60982.
Understanding when and where protein-protein interactions (PPIs) occur is critical to understanding protein function in the cell and how broader processes such as development are affected. The Caenorhabditis elegans germline is a great model system for studying PPIs that are related to the regulation of stem cells, meiosis, and development. There are a variety of well-developed techniques that allow proteins of interest to be tagged for recognition by standard antibodies, making this system advantageous for proximity ligation assay (PLA) reactions. As a result, the PLA is able to show where PPIs occur in a spatial and temporal manner in germlines more effectively than alternative approaches. Described here is a protocol for the application and quantification of this technology to probe PPIs in the C. elegans germline.
了解蛋白质-蛋白质相互作用（PPI）在何时何地发生，对于理解细胞中的蛋白质功能以及发育等更广泛的过程如何受到影响至关重要。秀丽隐杆线虫生殖系是研究与干细胞调控、减数分裂和发育相关的PPI的理想模型系统。有多种成熟的技术可将感兴趣的蛋白质进行标记，以便被标准抗体识别，这使得该系统有利于进行邻近连接分析（PLA）反应。因此，与其他方法相比，PLA能够更有效地在生殖系中以时空方式显示PPI发生的位置。本文介绍了一种将该技术应用于秀丽隐杆线虫生殖系中探测PPI并进行定量分析的方案。
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