Studies toward in vitro reconstitution of plant chromatin

WCU Author/Contributor (non-WCU co-authors, if there are any, appear on document)
Nicholas Martinez (Creator)
Institution
Western Carolina University (WCU )
Web Site: http://www.wcu.edu/404.asp
Advisor
Christopher Coburn

Abstract: 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.

Additional Information

Publication
Thesis
Language: English
Date: 2009
Subjects
Chromatin
Gene expression