Synchronization of Secretory Cargos Trafficking in Populations of Cells
Synchronization of Secretory Cargos Trafficking in Populations of Cells
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DOI：
10.1007/978-1-4939-0944-5_14
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发表时间：
2014-01-01
期刊：
影响因子：
--
通讯作者：
Perez, Franck
中科院分区：
文献类型：
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作者：
Boncompain, Gaelle;Perez, Franck
在哺乳动物细胞中，分泌蛋白通过分泌途径转运到其目的区室。货物在内质网中开始它们的旅程，然后到达高尔基复合体，在那里它们被加工和分选以被递送到它们的靶细胞内区室。为了分析和可视化这种蛋白质流动，人们需要同步它们的运输。我们最近开发了保留使用选择性钩（RUSH）系统，使同时和同步释放的分泌货物从内质网在一个人口的细胞。在这里，我们描述了如何将编码感兴趣的货物的基因亚克隆到RUSH质粒中，并在固定样品和活细胞中监测其沿分泌途径的同步运输沿着。
In mammalian cells, secretory proteins are transported to their destination compartment via the secretory pathway. Cargos start their journey in the endoplasmic reticulum and then reach the Golgi complex where they are processed and sorted to be delivered to their target intracellular compartment. To analyze and visualize this flow of proteins, one needs to synchronize their transport. We recently developed the retention using selective hooks (RUSH) system enabling simultaneous and synchronous release of secretory cargos from the endoplasmic reticulum in a population of cells. Here, we describe how to subclone the gene coding for a cargo of interest into a RUSH plasmid and to monitor its synchronized transport along the secretory pathway in fixed samples and in living cells.