Transient Expression of Fluorescent Fusion Proteins in Arabidopsis Protoplasts
| Autor: | Yonglun Zeng, Changyang Ji, Youshun Lin, Liwen Jiang |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine biology Chemistry fungi Cell Golgi apparatus biology.organism_classification Subcellular localization Plant cell 01 natural sciences Fusion protein Protein subcellular localization prediction Cell biology 03 medical and health sciences symbols.namesake 030104 developmental biology medicine.anatomical_structure Arabidopsis medicine symbols Endoplasmic reticulum exit site 010606 plant biology & botany |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071608791 |
| DOI: | 10.1007/978-1-0716-0880-7_7 |
| Popis: | Transient expression using protoplasts isolated from Arabidopsis suspension culture cells is a fast and useful tool for analyzing protein subcellular localization and dynamics in plant cells. Recently, super-resolution imaging techniques such as N-SIM (Nikon, Structured Illumination Microscopy) are widely used in cell biology study, allowing cell biologists to obtain unattainable details and relationships of cell structures and functions by conventional confocal imaging. To facilitate the usage of protoplasts transient expression and super-resolution imaging for protein localization and dynamic analysis in plant cell biology research, here we describe updated protocols of protoplasts isolation from Arabidopsis suspension culture cells and transient expression assay for protein trafficking and localization study. Further, using GFP-tagged ERES (Endoplasmic Reticulum Exit Site) marker proteins and RFP-tagged Golgi marker as examples, we illustrate the major tools and methods for protein localization analysis using super-resolution imaging. |
| Databáze: | OpenAIRE |
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