Abstract
Following the discovery that the green fluorescent protein (GFP) fluoresces when expressed in foreign organisms (1) and that it can serve as a reporter in living cells when fused to another protein (2), GFP has come into wide use. GFP offers advantages over other reporters in that it fluoresces in live cells and its fluorescence does not require the addition of any substrates, cofactors, or accessory proteins. This means that GFP-tagged proteins can be used to follow the dynamics of specific proteins in living cells, making it feasible to carry out experiments that previously were either difficult or not possible to perform. GFP fusion proteins can also be targeted for expression in given cellular compartments and it is possible to ensure that all of the protein is labeled by transferring the gene fusion into a null mutant, representing a great increase in efficiency compared to previous methods of fluorescent labeling of proteins. Moreover, GFP fluorescence is long-lasting, and its sensitivity to pH and temperature has enabled workers to use GFP fusion proteins to measure pH in specific cellular compartments (3) and to follow protein localization in temperature-shift experiments (4).
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Endow, S.A. (2001). Green Fluorescent Protein as a Tag for Molecular Motor Proteins. In: Vernos, I. (eds) Kinesin Protocols. Methods in Molecular Biology™, vol 164. Humana Press. https://doi.org/10.1385/1-59259-069-1:123
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DOI: https://doi.org/10.1385/1-59259-069-1:123
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