Mitogen-activated protein kinases (MAPK) are key regulatory elements in many processes. They are highly conserved throughout eukaryotes. In plants, MAPKs are involved in biotic and abiotic stress responses; they regulate cell division, cell growth, and also programmed cell death. In vivo visualization of MAPKs is crucial for understanding of their spatiotemporal organization. Cloning of MAPK-fluorescent protein fusions might present difficulties related to the preservation of protein-protein interactions essential for MAPK localization, interactions with upstream and downstream regulators, and finally substrate targeting. In this chapter we describe cloning of MAPKs in the flexible MultiSite Gateway ® cloning system followed by easy and quick testing of binary vectors by transient assays in Arabidopsis thaliana and Nicotiana benthamiana.
CITATION STYLE
Doskočilová, A., Luptovčiak, I., Smékalová, V., & Šamaj, J. (2014). Fluorescent protein tagging of arabidopsis MAPKs for in vivo localization studies. Methods in Molecular Biology, 1171, 131–145. https://doi.org/10.1007/978-1-4939-0922-3_11
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