For example, various transgenic mouse lines ( ) have been produced with GFP labels on proteins 13 retransforming such lines with genes encoding new photoconvertible proteins would require considerable time and expense. In addition to specific collections, many investigators with interests in particular proteins or subcellular locations have produced cell lines or organisms with GFP tags. In Arabidopsis, lines are available with GFP tags on transcription factors 11, with GFP as promoter-reporters 11 and with GFP labels on subcellular structures 12. In Drosophila, there is a collection ( ) in which GFP has tagged full-length endogenous proteins expressed from their endogenous loci or used in an enhancer trap 9, 10. For example, a collection of strains ( ) expressing full-length GFP-tagged proteins is available for yeast 7, 8. However, a number of collections of GFP-tagged organisms had been made before these proteins became available for use. The development of photoconvertible fluorescent proteins such as mEosFP, tdEosFP, Dendra, Dronpa, Kaede, KikGR, mOrange allows new questions to be asked concerning dynamic processes in living cells 1, 2, 3, 4, 5, 6. While genes encoding fluorescent proteins specifically designed for photoconversion will usually be advantageous when creating new transgenic lines, our method for photoconversion of GFP allows the use of existing GFP-tagged transgenic lines for studies of dynamic processes in living cells.įusions of Green Fluorescent Protein (GFP) and its derivatives are extremely valuable tools for examining gene expression, protein and RNA localization, protein-protein interactions, protein synthesis and degradation and organelle movement. We demonstrate its use in transgenic plant, Drosophila and mammalian cells in vivo. Here we describe a fast, localized and non-invasive method for GFP photoconversion from green to red. However, before genes encoding these fluorescent proteins were available, many proteins have already been labelled with GFP in transgenic cells a number of model organisms feature collections of GFP-tagged lines and organisms. Such proteins are finding application in following the dynamics of particular proteins or labelled organelles within the cell. A variety of fluorescent proteins have been identified that undergo shifts in spectral emission properties over time or once they are irradiated by ultraviolet or blue light.
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