Honors Theses
Date of Award
2015
Document Type
Undergraduate Thesis
Department
Chemistry and Biochemistry
First Advisor
Randy Wadkins
Relational Format
Dissertation/Thesis
Abstract
Recent studies have illustrated the presence of several secondary, non-B DNA structures that are associated with the promoter regions of several oncogenes, such as the intensely studied MYC gene. Two such secondary structures, G-quadruplexes and i-Motifs, have become topics of interest due to their prevalence within oncogenic promoters and their potential accessibility for future drug targeting due to their globular natures. Though fairly extensive research has been conducted to understand different factors affecting G-quadruplex formation and stability, similar studies for i-Motifs, particularly related to thermodynamics, are still evolving. One factor known to contribute to the additional energy required for duplex DNA to transition to its single-stranded form capable of forming i-Motifs is protein-DNA binding. This has been observed using EMSA, providing an estimated equilibrium binding constant. Therefore, other methods should be explored to acquire a truer equilibrium binding constant and determination of the energy contributed by protein binding to i-Motif DNA. This thesis explores the method of fluorescence anisotropy as an alternative way of examining the thermodynamic effects protein-DNA binding has on i-Motif stability by using a sequence within the c-MYC promoter and one domain of a Poly Cytosine Binding Protein. Because of the problems incurred throughout various experimental techniques, however, further studies are needed to explore the usefulness of fluorescence anisotropy with this particular protein and DNA sequence.
Recommended Citation
Nobile, Danna, "An Investigation of Protein-DNA Binding Characteristics by Fluorescence Anisotropy" (2015). Honors Theses. 248.
https://egrove.olemiss.edu/hon_thesis/248
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