Mitogen-Activated Protein Kinase (MAPK) cascade is one of the main signaling components mediating abiotic and biotic stress and hormone information in plants. Plant MAPK study has been impeded with a genetic approach using a long-term phenotypic analysis in spite of the transient nature of the protein kinase signaling. Arabidopsis leaf mesophyll protoplasts provide a versatile resource for diverse cell-based assays to acquire immediate molecular and biochemical responses with transient expression of MAPK cascade components of interests. Thus, it is an attractive tool for a high-throughput functional analysis of Arabidopsis MAPK cascade signaling. However, transient expression in Arabidopsis mesophyll protoplast (TEAMP) system requires mastered skills for protoplast preparation and handling to achieve steady and stable data. Here, we have described two analytical methods for MAPK cascade signaling using TEAMP system.
Arabidopsis leaf mesophyll protoplasts Transient expression Immunocomplex kinase assay Protein blot analysis Mitogen-Activated Protein Kinase cascade
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We thank Prof Jen Sheen for sharing information and encouragement on this manuscript preparation. Most of all, we deeply appreciate tremendous efforts to establish and improve the TEAMP system by many former and current members of Sheen lab at HMS/MGH and Yoo lab at Korea University. This work has been supported by grants from the National Research Foundation (NRF-2012R1A2A2A01005933), Korean RDA (PJ009106052013) and KOREA University to Sang-Dong Yoo.
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