Abstract
A system for the expression of recombinant histone proteins has been developed and it provides the possibility of manipulating the histone proteins by site-directed mutagenesis. This can be used to study the roles of specific regions of the histone proteins in chromatin function and to introduce amino acids at specific sites within nucleosomes for the attachment of reporter compounds. This chapter describes the use of this approach to attach compounds that facilitate the study of the dynamic properties of chromatin. Recombinant histone technology can also be used to provide a means of attaching fluorescent dyes. Techniques for the detection of fluorescent dyes have increased, enabling detection down to the level of single molecules. To date, the major use of fluorescent technology to study chromatin structure has stemmed from the expression of green or yellow fluorescent protein (GFP or YFP, respectively) histone fusions in vivo. These have provided a powerful system for the study of the dynamic properties of chromatin in vivo .
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