Lee Myoung Hui, Lee Yongjik, Hwang Inhwan
Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, South Korea.
Methods Mol Biol. 2013;1043:113-20. doi: 10.1007/978-1-62703-532-3_12.
In eukaryotic cells, a large number of proteins are transported to their final destination after translation by a process called intracellular trafficking. Transient gene expression, either in plant protoplasts or in specific plant tissues, is a fast, flexible, and reproducible approach to study the cellular function of proteins, protein subcellular localizations, and protein-protein interactions. Here we describe the general method of protoplast isolation, polyethylene glycol-mediated protoplast transformation and immunostaining of protoplast or intact root tissues for studying the localization of protein in Arabidopsis.
在真核细胞中,大量蛋白质在翻译后通过一种称为细胞内运输的过程被转运到它们的最终目的地。在植物原生质体或特定植物组织中进行的瞬时基因表达,是一种快速、灵活且可重复的方法,用于研究蛋白质的细胞功能、蛋白质亚细胞定位以及蛋白质-蛋白质相互作用。在这里,我们描述了用于研究拟南芥中蛋白质定位的原生质体分离、聚乙二醇介导的原生质体转化以及原生质体或完整根组织免疫染色的一般方法。