液泡靶向蛋白：亚细胞定位研究的来龙去脉

Vacuole-Targeted Proteins: Ins and Outs of Subcellular Localization Studies.

作者信息

Carqueijeiro Inês, Sepúlveda Liuda J, Mosquera Angela, Payne Richard, Corbin Cyrielle, Papon Nicolas, de Bernonville Thomas Dugé, Besseau Sébastien, Lanoue Arnaud, Glévarec Gaëlle, Clastre Marc, St-Pierre Benoit, Atehortùa Lucia, Giglioli-Guivarc'h Nathalie, O'Connor Sarah E, Oudin Audrey, Courdavault Vincent

机构信息

EA2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, Tours, France.

Laboratorio de Biotecnología, Sede de Investigación Universitaria, Universidad de Antioquia, Medellin, Colombia.

出版信息

Methods Mol Biol. 2018;1789:33-54. doi: 10.1007/978-1-4939-7856-4_4.

DOI:10.1007/978-1-4939-7856-4_4

PMID:29916070

Abstract

Accurate and efficient demonstrations of protein localizations to the vacuole or tonoplast remain strict prerequisites to decipher the role of vacuoles in the whole plant cell biology and notably in defence processes. In this chapter, we describe a reliable procedure of protein subcellular localization study through transient transformations of Catharanthus roseus or onion cells and expression of fusions with fluorescent proteins allowing minimizing artefacts of targeting.

摘要

准确且高效地证明蛋白质定位于液泡或液泡膜仍然是解读液泡在整个植物细胞生物学中，尤其是在防御过程中的作用的严格前提条件。在本章中，我们描述了一种可靠的蛋白质亚细胞定位研究方法，该方法通过长春花或洋葱细胞的瞬时转化以及与荧光蛋白融合表达，从而尽量减少靶向假象。