Abstract
Publisher Summary The dynamics of proteins can be monitored in living cells after tagging them with the green fluorescent protein (GFP). A construct encoding GFP fused to the protein of interest is expressed in a cell, so the resulting fluorescent hybrid can be seen directly. Mutant GFPs with altered fluorescence are available (e.g., enhanced cyan and yellow fluorescent proteins—ECFPs and EYFPs), and one—PAGFP—is photoactivated by irradiation with 413-nm light so its fluorescence (488-nm excitation) increases 100-fold. Therefore, it is possible to monitor histone and polymerase dynamics in real time, using these GFP tags and photobleaching techniques such as fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP). This chapter presents the studies on the dynamics of the core histones and RNA polymerase II to illustrate these techniques.
Published Version
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