Transient expression assays for quantifying signaling output.
Transient expression assays for quantifying signaling output.
复制标题
DOI：
10.1007/978-1-61779-809-2_16
复制
发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Sheen, Jen
中科院分区：
文献类型：
--
作者：
Niu, Yajie;Sheen, Jen
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.
