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mEosFP-based green-to-red photoconvertible subcellular probes for plants.
Plant Physiol 2010; 154(4):1573-87PP

Abstract

Photoconvertible fluorescent proteins (FPs) are recent additions to the biologists' toolbox for understanding the living cell. Like green fluorescent protein (GFP), monomeric EosFP is bright green in color but is efficiently photoconverted into a red fluorescent form using a mild violet-blue excitation. Here, we report mEosFP-based probes that localize to the cytosol, plasma membrane invaginations, endosomes, prevacuolar vesicles, vacuoles, the endoplasmic reticulum, Golgi bodies, mitochondria, peroxisomes, and the two major cytoskeletal elements, filamentous actin and cortical microtubules. The mEosFP fusion proteins are smaller than GFP/red fluorescent protein-based probes and, as demonstrated here, provide several significant advantages for imaging of living plant cells. These include an ability to differentially color label a single cell or a group of cells in a developing organ, selectively highlight a region of a cell or a subpopulation of organelles and vesicles within a cell for tracking them, and understanding spatiotemporal aspects of interactions between similar as well as different organelles. In addition, mEosFP probes introduce a milder alternative to fluorescence recovery after photobleaching, whereby instead of photobleaching, photoconversion followed by recovery of green fluorescence can be used for estimating subcellular dynamics. Most importantly, the two fluorescent forms of mEosFP furnish bright internal controls during imaging experiments and are fully compatible with cyan fluorescent protein, GFP, yellow fluorescent protein, and red fluorescent protein fluorochromes for use in simultaneous, multicolor labeling schemes. Photoconvertible mEosFP-based subcellular probes promise to usher in a much higher degree of precision to live imaging of plant cells than has been possible so far using single-colored FPs.

Authors+Show Affiliations

Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada. jmathur@uoguelph.caNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20940350

Citation

Mathur, Jaideep, et al. "MEosFP-based Green-to-red Photoconvertible Subcellular Probes for Plants." Plant Physiology, vol. 154, no. 4, 2010, pp. 1573-87.
Mathur J, Radhamony R, Sinclair AM, et al. MEosFP-based green-to-red photoconvertible subcellular probes for plants. Plant Physiol. 2010;154(4):1573-87.
Mathur, J., Radhamony, R., Sinclair, A. M., Donoso, A., Dunn, N., Roach, E., ... Mathur, N. (2010). MEosFP-based green-to-red photoconvertible subcellular probes for plants. Plant Physiology, 154(4), pp. 1573-87. doi:10.1104/pp.110.165431.
Mathur J, et al. MEosFP-based Green-to-red Photoconvertible Subcellular Probes for Plants. Plant Physiol. 2010;154(4):1573-87. PubMed PMID: 20940350.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - mEosFP-based green-to-red photoconvertible subcellular probes for plants. AU - Mathur,Jaideep, AU - Radhamony,Resmi, AU - Sinclair,Alison M, AU - Donoso,Ana, AU - Dunn,Natalie, AU - Roach,Elyse, AU - Radford,Devon, AU - Mohaghegh,P S Mohammad, AU - Logan,David C, AU - Kokolic,Ksenija, AU - Mathur,Neeta, Y1 - 2010/10/12/ PY - 2010/10/14/entrez PY - 2010/10/14/pubmed PY - 2011/6/3/medline SP - 1573 EP - 87 JF - Plant physiology JO - Plant Physiol. VL - 154 IS - 4 N2 - Photoconvertible fluorescent proteins (FPs) are recent additions to the biologists' toolbox for understanding the living cell. Like green fluorescent protein (GFP), monomeric EosFP is bright green in color but is efficiently photoconverted into a red fluorescent form using a mild violet-blue excitation. Here, we report mEosFP-based probes that localize to the cytosol, plasma membrane invaginations, endosomes, prevacuolar vesicles, vacuoles, the endoplasmic reticulum, Golgi bodies, mitochondria, peroxisomes, and the two major cytoskeletal elements, filamentous actin and cortical microtubules. The mEosFP fusion proteins are smaller than GFP/red fluorescent protein-based probes and, as demonstrated here, provide several significant advantages for imaging of living plant cells. These include an ability to differentially color label a single cell or a group of cells in a developing organ, selectively highlight a region of a cell or a subpopulation of organelles and vesicles within a cell for tracking them, and understanding spatiotemporal aspects of interactions between similar as well as different organelles. In addition, mEosFP probes introduce a milder alternative to fluorescence recovery after photobleaching, whereby instead of photobleaching, photoconversion followed by recovery of green fluorescence can be used for estimating subcellular dynamics. Most importantly, the two fluorescent forms of mEosFP furnish bright internal controls during imaging experiments and are fully compatible with cyan fluorescent protein, GFP, yellow fluorescent protein, and red fluorescent protein fluorochromes for use in simultaneous, multicolor labeling schemes. Photoconvertible mEosFP-based subcellular probes promise to usher in a much higher degree of precision to live imaging of plant cells than has been possible so far using single-colored FPs. SN - 1532-2548 UR - https://www.unboundmedicine.com/medline/citation/20940350/mEosFP_based_green_to_red_photoconvertible_subcellular_probes_for_plants_ L2 - http://www.plantphysiol.org/cgi/pmidlookup?view=long&pmid=20940350 DB - PRIME DP - Unbound Medicine ER -