Date of Graduation

Spring 2018

Degree

Master of Science in Cell and Molecular Biology

Department

Biomedical Sciences

Committee Chair

Joshua Smith

Keywords

RAD23, RAD4, ubiquitin, nucleotide excision repair, proteasome, Tetrahymena thermophila

Subject Categories

Cell Biology | Molecular Biology | Molecular Genetics

Abstract

Rad23 is a protein involved in both nucleotide excision repair (NER) and proteasome-mediated degradation, and has been suggested to facilitate interactions between these two pathways. The model organism Tetrahymena thermophila, which has a transcriptionally silent micronucleus, provides a useful platform for studying the role of Rad23 in global genome NER (GG-NER). However, the ectopic expression systems used thus far in T. thermophila to study Rad23 are repressed by UV light and do not account for the background expression of endogenous RAD23; these phenomena prevent insightful gains to the true dynamics of Rad23. In this thesis, endogenous tagging cassettes have been designed to allow for the tagging of endogenous RAD23 or any other T. thermophila gene to circumvent the issues inherent to ectopic expression. Additionally, a plasmid has been made to facilitate the genetic knockout of RAD23 in T. thermophila. Basic phylogeny and expression analysis of RAD23 were also performed to better characterize this protein in T. thermophila. The tools designed in this study will aid future researchers in the genetic manipulation of T. thermophila.

Copyright

© Evan Andrew Wilson

Open Access

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