University of Illinois Urbana-Champaign

In Vitro Selection and Metal Specificity of Transition Metal-Dependent DNAzymes

Nelson, Kevin Eric

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Description

Title
In Vitro Selection and Metal Specificity of Transition Metal-Dependent DNAzymes
Author(s)
Nelson, Kevin Eric
Issue Date
2006
Doctoral Committee Chair(s)
Lu, Yi
Department of Study
Biochemistry
Discipline
Biochemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Language
eng
Abstract
The characterization of two families of Co2+-dependent DNAzymes further demonstrates the interplay between DNAzyme structure and metal specificity. The clone 11 and 18 DNAzymes were selected for study because the primary sequences differ by only four bases, producing vast differences in Co2+ specificity. Phylogenetic and mutational analyses were used to identify the sequence and secondary structure responsible for Co 2+ specificity and catalysis. Sequence elements within the random domain and conserved primer binding regions are important in stabilizing structures with high specificity and activity. Truncation of peripheral sequences leads to an increased propensity for formation of inactive, self-complementary or metastable structures. The Co2+ ion likely stabilizes a conformation with high activity and specificity in the clone 11 DNAzyme system. The aspects of DNA/RNAzyme function and metal specificity discussed will likely aid in the rational design or in vitro selection of DNA/RNAzyme systems well suited for the spectroscopic characterization of metal binding sites and showing greater potential for therapeutic and biotechnology applications.
Graduation Semester
2006
Type of Resource
text
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http://hdl.handle.net/2142/84826

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