Transient expression assays for quantifying signaling output

Methods in Molecular Biology
Yajie Niu, Jen Sheen

Abstract

The protoplast transient expression system has become a powerful and popular tool for studying molecular mechanisms underlying various plant signal transduction pathways. Arabidopsis mesophyll protoplasts display intact and active physiological responses and are easy to isolate and transfect, which facilitate high-throughput screening and systematic and genome-wide characterization of gene functions. The system is suitable for most Arabidopsis accessions and mutant plants. Genetic complementation of mutant defective in sensor functions, gene expression, enzymatic activities, protein interactions, and protein trafficking can be easily designed and explored in cell-based assays. Here, we describe the detailed protocols for protoplast isolation, polyethylene glycol-calcium transfection, and different assays for quantifying the output of various signaling pathways.

Citations

May 18, 2013·Journal of Experimental Botany·Ive De SmetGerd Jürgens
Jan 28, 2014·The Arabidopsis Book·Joseph J Kieber, G Eric Schaller
Apr 5, 2013·The Plant Journal : for Cell and Molecular Biology·Matthew RamonFilip Rolland
Nov 9, 2016·Plant Methods·Zhicheng ZhangKoen Geuten
Aug 5, 2017·The Plant Journal : for Cell and Molecular Biology·Javier CanalesRodrigo A Gutiérrez

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