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A Study of DNA Replication and Repair Proteins from Bacteriophage T4 and a Related Phage

Senger, Anne Benedict
http://rave.ohiolink.edu/etdc/view?acc_num=toledo1104776177

Abstract Details

2004, Master of Science, University of Toledo, Chemistry.
In the Mueser laboratory, we study how DNA replication and repair proteins recognize DNA in a structure-specific manner. Bacteriophage T4 is used as a model system to study DNA replication as it encodes all ten proteins required for DNA replication. Much is known about how the individual proteins function in replication but not much is known about the structural aspects of the protein-protein or protein-DNA interactions at the replication fork. The goal of our research is to study how these replication proteins interact with each other and with DNA. We work towards achieving this goal by crystallizing the protein-protein and protein-DNA complexes and then solving their structures, using macromolecular crystallography techniques. We then use the structural information gathered to analyze the interactions. The overall goal of this master’s thesis project was to learn many of the techniques involved in protein chemistry and protein crystallization. My research was tailored to protein expression, purification and crystallization so I could learn an array of techniques and become familiar with various pieces of instrumentation. I wanted to be able to use this knowledge in future research positions. My work was focused on two of the replication proteins from Bacteriophage T4: T4 gene 59 helicase assembly protein and T4 gene 32 single-stranded binding protein. These two proteins interact in the absence of DNA and form a complex at the replication fork. I was responsible for expressing mutated and truncated forms of the native proteins on a large scale and developing purification protocols in order to prepare pure protein for crystal screening. After my research with the T4 helicase assembly protein began, I also started working on a similar helicase assembly protein from a related system – bacteriophage KVP40 59 protein. I was also responsible for developing a purification protocol for single-stranded DNA substrates that were used to prepare forked substrates for the crystal screening.
Timothy Mueser (Advisor)
139 p.
Chemistry, Biochemistry
DNA replication; bacteriophage T4; bacteriophage KVP40; helicase assembly; single-stranded binding; protein crystallization; DNA purification

Recommended Citations

Citations

  • Senger, A. B. (2004). A Study of DNA Replication and Repair Proteins from Bacteriophage T4 and a Related Phage [Master's thesis, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1104776177

    APA Style (7th edition)

  • Senger, Anne. A Study of DNA Replication and Repair Proteins from Bacteriophage T4 and a Related Phage. 2004. University of Toledo, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=toledo1104776177.

    MLA Style (8th edition)

  • Senger, Anne. "A Study of DNA Replication and Repair Proteins from Bacteriophage T4 and a Related Phage." Master's thesis, University of Toledo, 2004. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1104776177

    Chicago Manual of Style (17th edition)

toledo1104776177
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© 2004, all rights reserved.
This open access ETD is published by University of Toledo and OhioLINK.