title: Fluorescent protein tagging of Arabidopsis MAPKs for in vivo localization studies.
abstract: 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.
authors.affiliation: Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University Olomouc, Šlechtitelů 11, 783 71, Olomouc, Czech Republic, anna.doskocilova@upol.cz.
authors.forename: Anna
authors.initials: A
authors.lastname: Doskočilová
authors.forename: Ivan
authors.initials: I
authors.lastname: Luptovčiak
authors.forename: Veronika
authors.initials: V
authors.lastname: Smékalová
authors.forename: Jozef
authors.initials: J
authors.lastname: Samaj
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism*; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D018745:Genome, Plant / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics*; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D018515:Plant Leaves / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D029681:Arabidopsis Proteins; D002593:Cetrimonium Compounds; D017931:DNA Primers; D018744:DNA, Plant; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C436293:MPK6 protein, Arabidopsis; D020928:Mitogen-Activated Protein Kinases; D000077286:Cetrimonium