Degree Type

Thesis

Date of Award

2016

Degree Name

Master of Science

Department

Biochemistry, Biophysics and Molecular Biology

Major

Biochemistry

First Advisor

Michael Shogren-Knaak

Abstract

Chromatin, the native form of eukaryotic DNA, organizes the genome and regulates its use. Assembled chromatin model systems provide a powerful tool for understanding the complex structural and functional properties of chromatin. Chromatin is structurally diverse within the cell, yet current in vitro model systems do not reflect this diversity. Additionally, generating new model systems has traditionally been challenging. Here we apply the ideas of solid-phase synthesis to show that sequential ligation of DNA fragments readily generates a wide range of chromatin model systems. Using DNA fragments containing one or more nucleosome positioning sites, we generated DNA templates for assembling nucleosomal arrays containing from one to forty-eight nucleosomes, where the spacing between nucleosomes was also varied. We successfully demonstrate that the nucleosomal arrays generated are well saturated and compositionally well defined. Altogether, these strategies provide a means to vary the length and composition of chromatin models systems at the level of individual nucleosomes, and provide new tool for probing chromatin structure and function.

Copyright Owner

Margaret K. Gannon

Language

en

File Format

application/pdf

File Size

38 pages

Share

COinS