Abstract
In eukaryotic cells, a large number of proteins are transported to their final destination after translation by a process called intracellular trafficking. Transient gene expression, either in plant protoplasts or in specific plant tissues, is a fast, flexible, and reproducible approach to study the cellular function of proteins, protein subcellular localizations, and protein–protein interactions. Here we describe the general method of protoplast isolation, polyethylene glycol-mediated protoplast transformation and immunostaining of protoplast or intact root tissues for studying the localization of protein in Arabidopsis.
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References
Juergens G (2004) Membrane trafficking in plants. Annu Rev Cell Dev Biol 20:481–504
Hwang I, Robinson DG (2009) Transport vesicle formation in plant cells. Curr Opin Plant Biol 12:660–669
Bröcker C, Engelbrecht-Vandré S, Ungermann C (2010) Multisubunit tethering complexes and their role in membrane fusion. Curr Biol 20:943–952
Nielsen E, Cheung AY, Ueda T (2008) The regulatory RAB and ARF GTPases for vesicular trafficking. Plant Physiol 147:1516–1526
Parinov S, Sundaresan V (2000) Functional genomics in Arabidopsis: large-scale insertional mutagenesis complements the genome sequencing project. Curr Opin Biotechnol 11:157–161
An G, Jeong DH, Jung KH, Lee S (2005) Reverse genetic approaches for functional genomics of rice. Plant Mol Biol 59:111–123
Denecke J, Aniento F, Frigerio L, Hawes C, Hwang I, Mathur J, Neuhaus JM, Robinson DG (2012) Secretory pathway research: the more experimental systems the better. Plant Cell 24:1316–1326
Tsuda K, Qi Y, le Nguyen V, Bethke G, Tsuda Y, Glazebrook J, Katagiri F (2012) An efficient Agrobacterium-mediated transient transformation of Arabidopsis. Plant J 69:713–719
Van Loock B, Markakis MN, Verbelen JP, Vissenberg K (2010) High-throughput transient transformation of Arabidopsis roots enables systematic colocalization analysis of GFP-tagged proteins. Plant Signal Behav 5:261–263
Jung CJ, Lee MH, Min MK, Hwang I (2011) Localization and trafficking of an isoform of the AtPRA1 family to the Golgi apparatus depend on both N- and C-terminal sequence motifs. Traffic 12:185–200
Chen S, Tao L, Zeng L, Vega-Sanchez ME, Umemura K, Wang GL (2006) A highly efficient transient protoplast system for analyzing defence gene expression and protein-protein interactions in rice. Mol Plant Pathol 7:417–427
Sheen J (2001) Signal transduction in maize and Arabidopsis mesophyll protoplasts. Plant Physiol 127:1466–1475
Reyes FC, Buono R, Otegui MS (2011) Plant endosomal trafficking pathways. Curr Opin Plant Biol 14:666–673
Hwang I (2008) Sorting and anterograde trafficking at the Golgi apparatus. Plant Physiol 148:673–683
Emanuelsson O, Brunak S, von Heijne G, Nielsen H (2007) Locating proteins in the cell using TargetP, SignalP and related tools. Nat Protoc 2:953–971
Wee EG, Sherrier DJ, Prime TA, Dupree P (1998) Targeting of active sialyltransferase to the plant Golgi apparatus. Plant Cell 10:1759–1768
Kircher S, Wellmer F, Nick P, Rügner A, Schäfer E, Harter K (1999) Nuclear import of the parsley bZIP transcription factor CPRF2 is regulated by phytochrome photoreceptors. J Cell Biol 144:201–211
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Lee, M.H., Lee, Y., Hwang, I. (2013). In Vivo Localization in Arabidopsis Protoplasts and Root Tissue. In: Running, M. (eds) G Protein-Coupled Receptor Signaling in Plants. Methods in Molecular Biology, vol 1043. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-532-3_12
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DOI: https://doi.org/10.1007/978-1-62703-532-3_12
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