Department of Biology, University of Crete, Heraklion, Greece.
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Heraklion, Greece.
Methods Mol Biol. 2022;2382:233-243. doi: 10.1007/978-1-0716-1744-1_14.
Membrane trafficking is central to cell plate construction during plant cytokinesis. Studies on cell plate formation can provide answers to basic biology questions including molecular mechanisms of membrane trafficking, tissue patterning, and cytoskeletal dynamics. Consequently, a detailed understanding of cytokinesis depends on the characterization of molecules that function in the formation, transport, targeting, and fusion of membrane vesicles and delivery of proteins to the developing and maturing plate. This chapter describes a pipeline based on fluorescence recovery after photobleaching (FRAP) to measure and analyze turnover of peripheral or transmembrane proteins on the cell plate. The approach described here can also be applied in other biological contexts.
膜运输在植物胞质分裂过程中的细胞板形成中起着核心作用。对细胞板形成的研究可以为基本生物学问题提供答案，包括膜运输、组织模式形成和细胞骨架动力学的分子机制。因此，对胞质分裂的详细了解取决于对在膜小泡的形成、运输、靶向和融合以及将蛋白质递送至发育和成熟的细胞板中起作用的分子的特征化。本章描述了一个基于光漂白后荧光恢复（FRAP）的流水线，用于测量和分析细胞板上外周或跨膜蛋白的周转率。这里描述的方法也可以应用于其他生物学背景。
