Title |
Targeted Green-Red Photoconversion of EosFP, a Fluorescent Marker Protein
|
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Published in |
Journal of Biological Physics, December 2005
|
DOI | 10.1007/s10867-005-0174-z |
Pubmed ID | |
Authors |
Sergey Ivanchenko, Carlheinz Röcker, Franz Oswald, Jörg Wiedenmann, G. Ulrich Nienhaus |
Abstract |
EosFP is a novel fluorescent protein from the stony coral Lobophyllia hemprichii. Its gene was cloned in Escherichia coli to express the tetrameric wild-type protein. The protein emits strong green fluorescence (516 nm) that shifts toward red (581 nm) upon near-ultraviolet irradiation at ∼390 nm due to a photo-induced modification that involves a break in the peptide backbone next to the chromophore. Using site-directed mutagenesis, dimeric (d1EosFP, d2EosFP) and monomeric (mEosFP) variants were produced with essentially unaltered spectroscopic properties. Here we present a spectroscopic characterization of EosFP and its variants, including room- and low-temperature spectra, fluorescence lifetime determinations, two-photon excitation and two-photon photoconversion. Furthermore, by transfection of a human cancer (HeLa) cell with a fusion construct of a mitochondrial targeting sequence and d2EosFP, we demonstrate how localized photoconversion of EosFP can be employed for resolving intracellular processes. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 55 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 11 | 20% |
Student > Bachelor | 11 | 20% |
Researcher | 8 | 15% |
Student > Doctoral Student | 6 | 11% |
Student > Master | 5 | 9% |
Other | 7 | 13% |
Unknown | 7 | 13% |
Readers by discipline | Count | As % |
---|---|---|
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Biochemistry, Genetics and Molecular Biology | 7 | 13% |
Chemistry | 7 | 13% |
Medicine and Dentistry | 5 | 9% |
Physics and Astronomy | 3 | 5% |
Other | 9 | 16% |
Unknown | 7 | 13% |