Chemistry, Department of
Department of Chemistry: Dissertations, Theses, and Student Research
First Advisor
Carolyn Mary Price
Date of this Version
11-1994
Document Type
Thesis
Citation
A thesis presented to the faculty of the Graduate College in the University of Nebraska in partial fulfillment of requirements for the degree of Master of Science
Major: Chemistry
Under the supervision of Professor Carolyn Mary Price
Lincoln, Nebraska, November 1994
Abstract
A novel PCR technique used to amplify Euplotes crassus macronuclear chromosomes was developed to provide scientific proof that an Euplotes crassus 1.0 kb gene is not genetically related to an Oxytricha nova 1.8 kb β-telomere protein gene. Theories and experimental procedures associated with the development of this PCR technique were the product of research that will have a profound impact on future studies of telomere protein genes and telomeric DNA in a multitude of eukaryotic cells. The hypothesis that a β-telomere protein does not exist in Euplotes crassus was supported by the result of this study.
Expression of the recombinant Euplotes crassus α-telomere protein in E. coli was achieved. The solubilization problem associated with the Euplotes crassus α-telomere protein, that has perplexed scientists for many years, was accomplished by taking advantage of the physiochemical properties of the α-telomere protein molecule. Characterization of solubilized recombinant Euplotes crassus α-telomere protein has provided a better understanding of the molecular properties associated with the function of this protein. The basic properties of the Euplotes crassus α-telomere protein were shown to play an important role in the formation of higher ordered telomere protein DNA complexes in vitro. A relationship between the behavior and function of the Euplotes crassus α-telomere protein in solution has provided insight to the understanding of how the α-telomere protein functions in vivo to protect the ends of chromosomes. Construction of a unique bacterial overexpression vector that will be used to study mutants of the Euplotes crassus α-telomere protein was also performed. Ultimately, a better understanding of the physical and chemical prope,r ties of the Euplotes crassus α-telomere protein, that serve to protect the ends of macronuclear chromosomes, has resulted from this study.
Advisor: Carolyn Mary Price
Included in
Biochemistry Commons, Genetic Structures Commons, Molecular Genetics Commons, Organic Chemistry Commons
Comments
Copyright 1994, John Scott Perez. Used by permission