To study the relation between chromatin structure and DNA function in detail it is
necessary to have an in vitro procedure for assembling nucleosomes on a naked DNA
template with properties similar to native chromatin. Such procedures exist for yeast and
animal model systems but have not been developed for plants. The goal of this project
was to lay the groundwork for developing a chromatin assembly extract from plants.
Extracts from various plant materials were tested to determine their suitability for
chromatin reconstitution. Tissues from plants are thought to have much higher levels of
protease and nuclease activities than those of animals or yeast. Therefore, methods to
determine the relative activity of proteases and nucleases had to be developed to
determine if the template DNA, histones, and chromatin assembly proteins could survive
the chromatin assembly reaction. Additionally, methods to streamline the isolation of
maize nuclei and purification of histones were developed. This work lays the foundation
for future research that could result in extracts to reconstitute plant chromatin in vitro
