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mEosFP-based green-to-red photoconvertible subcellular probes for plants.

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.

journal_name

Plant Physiol

journal_title

Plant physiology

authors

Mathur J,Radhamony R,Sinclair AM,Donoso A,Dunn N,Roach E,Radford D,Mohaghegh PS,Logan DC,Kokolic K,Mathur N

doi

10.1104/pp.110.165431

subject

Has Abstract

pub_date

2010-12-01 00:00:00

pages

1573-87

issue

eissn

0032-0889

issn

1532-2548

pii

pp.110.165431

journal_volume

154

pub_type

杂志文章

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mEosFP-based green-to-red photoconvertible subcellular probes for plants.