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基于荧光蛋白的植物亚细胞蛋白定位方法。

Fluorescent Protein-Based Approaches for Subcellular Protein Localization in Plants.

机构信息

Institute of Molecular Plant Biology, ETH Zürich, Zürich, Switzerland.

Institute of Biology-Plant Physiology, Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.

出版信息

Methods Mol Biol. 2023;2564:203-211. doi: 10.1007/978-1-0716-2667-2_9.

Abstract

Fluorescent proteins (FPs) remarkably advanced the study of cellular biology of plants. The most common application is their use as reporter proteins to determine the subcellular localization of a protein of interest (POI) by endogenous expression of a suitable FP-POI fusion construct in plant cells. In this chapter we describe three approaches, namely, particle bombardment, protoplast transformation, and Agrobacterium infiltration, to transiently express such fusion constructs in plant cells of different species. These approaches are versatile and can be utilized for diverse fluorescent protein-based applications.

摘要

荧光蛋白(FPs)极大地推动了植物细胞生物学的研究。最常见的应用是将其用作报告蛋白,通过在植物细胞中内源表达合适的 FP-POI 融合构建体来确定感兴趣的蛋白质(POI)的亚细胞定位。在本章中,我们描述了三种方法,即粒子轰击、原生质体转化和农杆菌浸润,用于在不同物种的植物细胞中瞬时表达这种融合构建体。这些方法具有通用性,可以用于各种基于荧光蛋白的应用。

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