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- Genre: Doctoral Dissertation
- Creators: Campbell, Andrew
- Creators: Yan, Hao
Description
The principle of Darwinian evolution has been applied in the laboratory to nucleic acid molecules since 1990, and led to the emergence of in vitro evolution technique. The methodology of in vitro evolution surveys a large number of different molecules simultaneously for a pre-defined chemical property, and enrich for molecules with the particular property. DNA and RNA sequences with versatile functions have been identified by in vitro selection experiments, but many basic questions remain to be answered about how these molecules achieve their functions. This dissertation first focuses on addressing a fundamental question regarding the molecular recognition properties of in vitro selected DNA sequences, namely whether negatively charged DNA sequences can be evolved to bind alkaline proteins with high specificity. We showed that DNA binders could be made, through carefully designed stringent in vitro selection, to discriminate different alkaline proteins. The focus of this dissertation is then shifted to in vitro evolution of an artificial genetic polymer called threose nucleic acid (TNA). TNA has been considered a potential RNA progenitor during early evolution of life on Earth. However, further experimental evidence to support TNA as a primordial genetic material is lacking. In this dissertation we demonstrated the capacity of TNA to form stable tertiary structure with specific ligand binding property, which suggests a possible role of TNA as a pre-RNA genetic polymer. Additionally, we discussed the challenges in in vitro evolution for TNA enzymes and developed the necessary methodology for future TNA enzyme evolution.
ContributorsYu, Hanyang (Author) / Chaput, John C (Thesis advisor) / Chen, Julian (Committee member) / Yan, Hao (Committee member) / Arizona State University (Publisher)
Created2013
Description
This document is intended to show the various kinds of stylistically appropriate melodic and rhythmic ornamentation that can be used in the improvisation of the Sarabandes by J.S. Bach. Traditional editions of Bach's and other Baroque-era keyboard works have reflected evolving historical trends. The historical performance movement and other attempts to "clean up" pre-1950s romanticized performances have greatly limited the freedom and experimentation that was the original intention of these dances. Prior to this study, few ornamented editions of these works have been published. Although traditional practices do not necessarily encourage classical improvisation in performance I argue that manipulation of the melodic and rhythmic layers over the established harmonic progressions will not only provide diversity within the individual dance movements, but also further engage the ears of the performer and listener which encourages further creative exploration. I will focus this study on the ornamentation of all six Sarabandes from J.S. Bach's French Suites and show how various types of melodic and rhythmic variation can provide aurally pleasing alternatives to the composed score without disrupting the harmonic fluency. The author intends this document to be used as a pedagogical tool and the fully ornamented Sarabandes from J.S. Bach's French Suites are included with this document.
ContributorsOakley, Ashley (Author) / Meir, Baruch (Thesis advisor) / Campbell, Andrew (Committee member) / Norton, Kay (Committee member) / Pagano, Caio (Committee member) / Ryan, Russell (Committee member) / Arizona State University (Publisher)
Created2013
Description
The ribosome is a ribozyme and central to the biosynthesis of proteins in all organisms. It has a strong bias against non-alpha-L-amino acids, such as alpha-D-amino acids and beta-amino acids. Additionally, the ribosome is only able to incorporate one amino acid in response to one codon. It has been demonstrated that reengineering of the peptidyltransferase center (PTC) of the ribosome enabled the incorporation of both alpha-D-amino acids and beta-amino acids into full length protein. Described in Chapter 2 are five modified ribosomes having modifications in the peptidyltrasnferase center in the 23S rRNA. These modified ribosomes successfully incorporated five different beta-amino acids (2.1 - 2.5) into E. coli dihydrofolate reductase (DHFR). The second project (Chapter 3) focused on the study of the modified ribosomes facilitating the incorporation of the dipeptide glycylphenylalanine (3.25) and fluorescent dipeptidomimetic 3.26 into DHFR. These ribosomes also had modifications in the peptidyltransferase center in the 23S rRNA of the 50S ribosomal subunit. The modified DHFRs having beta-amino acids 2.3 and 2.5, dipeptide glycylphenylalanine (3.25) and dipeptidomimetic 3.26 were successfully characterized by the MALDI-MS analysis of the peptide fragments produced by "in-gel" trypsin digestion of the modified proteins. The fluorescent spectra of the dipeptidomimetic 3.26 and modified DHFR having fluorescent dipeptidomimetic 3.26 were also measured. The type I and II DNA topoisomerases have been firmly established as effective molecular targets for many antitumor drugs. A "classical" topoisomerase I or II poison acts by misaligning the free hydroxyl group of the sugar moiety of DNA and preventing the reverse transesterfication reaction to religate DNA. There have been only two classes of compounds, saintopin and topopyrones, reported as dual topoisomerase I and II poisons. Chapter 4 describes the synthesis and biological evaluation of topopyrones. Compound 4.10, employed at 20 µM, was as efficient as 0.5 uM camptothecin, a potent topoisomerase I poison, in stabilizing the covalent binary complex (~30%). When compared with a known topoisomerase II poison, etoposide (at 0.5 uM), topopyorone 4.10 produced similar levels of stabilized DNA-enzyme binary complex (~34%) at 5 uM concentration.
ContributorsMaini, Rumit (Author) / Hecht, Sidney M. (Thesis advisor) / Gould, Ian (Committee member) / Yan, Hao (Committee member) / Arizona State University (Publisher)
Created2013
Description
The biological and chemical diversity of protein structure and function can be greatly expanded by position-specific incorporation of non-natural amino acids bearing a variety of functional groups. Non-cognate amino acids can be incorporated into proteins at specific sites by using orthogonal aminoacyl-tRNA synthetase/tRNA pairs in conjunction with nonsense, rare, or 4-bp codons. There has been considerable progress in developing new types of amino acids, in identifying novel methods of tRNA aminoacylation, and in expanding the genetic code to direct their position. Chemical aminoacylation of tRNAs is accomplished by acylation and ligation of a dinucleotide (pdCpA) to the 3'-terminus of truncated tRNA. This strategy allows the incorporation of a wide range of natural and unnatural amino acids into pre-determined sites, thereby facilitating the study of structure-function relationships in proteins and allowing the investigation of their biological, biochemical and biophysical properties. Described in Chapter 1 is the current methodology for synthesizing aminoacylated suppressor tRNAs. Aminoacylated suppressor tRNACUAs are typically prepared by linking pre-aminoacylated dinucleotides (aminoacyl-pdCpAs) to 74 nucleotide (nt) truncated tRNAs (tRNA-COH) via a T4 RNA ligase mediated reaction. Alternatively, there is another route outlined in Chapter 1 that utilizes a different pre-aminoacylated dinucleotide, AppA. This dinucleotide has been shown to be a suitable substrate for T4 RNA ligase mediated coupling with abbreviated tRNA-COHs for production of 76 nt aminoacyl-tRNACUAs. The synthesized suppressor tRNAs have been shown to participate in protein synthesis in vitro, in an S30 (E. coli) coupled transcription-translation system in which there is a UAG codon in the mRNA at the position corresponding to Val10. Chapter 2 describes the synthesis of two non-proteinogenic amino acids, L-thiothreonine and L-allo-thiothreonine, and their incorporation into predetermined positions of a catalytically competent dihydrofolate reductase (DHFR) analogue lacking cysteine. Here, the elaborated proteins were site-specifically derivitized with a fluorophore at the thiothreonine residue. The synthesis and incorporation of phosphorotyrosine derivatives into DHFR is illustrated in Chapter 3. Three different phosphorylated tyrosine derivatives were prepared: bis-nitrobenzylphosphoro-L-tyrosine, nitrobenzylphosphoro-L-tyrosine, and phosphoro-L-tyrosine. Their ability to participate in a protein synthesis system was also evaluated.
ContributorsNangreave, Ryan Christopher (Author) / Hecht, Sidney M. (Thesis advisor) / Yan, Hao (Committee member) / Gould, Ian (Committee member) / Arizona State University (Publisher)
Created2013
Description
Solution conformations and dynamics of proteins and protein-DNA complexes are often difficult to predict from their crystal structures. The crystal structure only shows a snapshot of the different conformations these biological molecules can have in solution. Multiple different conformations can exist in solution and potentially have more importance in the biological activity. DNA sliding clamps are a family of proteins with known crystal structures. These clamps encircle the DNA and enable other proteins to interact more efficiently with the DNA. Eukaryotic PCNA and prokaryotic β clamp are two of these clamps, some of the most stable homo-oligomers known. However, their solution stability and conformational equilibrium have not been investigated in depth before. Presented here are the studies involving two sliding clamps: yeast PCNA and bacterial β clamp. These studies show that the β clamp has a very different solution stability than PCNA. These conclusions were reached through various different fluorescence-based experiments, including fluorescence correlation spectroscopy (FCS), Förster resonance energy transfer (FRET), single molecule fluorescence, and various time resolved fluorescence techniques. Interpretations of these, and all other, fluorescence-based experiments are often affected by the properties of the fluorophores employed. Often the fluorescence properties of these fluorophores are influenced by their microenvironments. Fluorophores are known to sometimes interact with biological molecules, and this can have pronounced effects on the rotational mobility and photophysical properties of the dye. Misunderstanding the effect of these photophysical and rotational properties can lead to a misinterpretation of the obtained data. In this thesis, photophysical behaviors of various organic dyes were studied in the presence of deoxymononucleotides to examine more closely how interactions between fluorophores and DNA bases can affect fluorescent properties. Furthermore, the properties of cyanine dyes when bound to DNA and the effect of restricted rotation on FRET are presented in this thesis. This thesis involves studying fluorophore photophysics in various microenvironments and then expanding into the solution stability and dynamics of the DNA sliding clamps.
ContributorsRanjit, Suman (Author) / Levitus, Marcia (Thesis advisor) / Lindsay, Stuart (Committee member) / Yan, Hao (Committee member) / Arizona State University (Publisher)
Created2013
Description
ABSTRACT Musicians endure injuries at an alarming rate, largely due to the misuse of their bodies. Musicians move their bodies for a living and therefore should understand how to move them in a healthy way. This paper presents Body Mapping as an injury prevention technique specifically directed toward collaborative pianists. A body map is the self-representation in one's brain that includes information on the structure, function, and size of one's body; Body Mapping is the process of refining one's body map to produce coordinated movement. In addition to preventing injury, Body Mapping provides a means to achieve greater musical artistry through the training of movement, attention, and the senses. With the main function of collaborating with one or more musical partners, a collaborative pianist will have the opportunity to share the knowledge of Body Mapping with many fellow musicians. This study demonstrates the author's credentials as a Body Mapping instructor, the current status of the field of collaborative piano, and the recommendation for increased body awareness. Information on the nature and abundance of injuries and Body Mapping concepts are also analyzed. The study culminates in a course syllabus entitled An Introduction to Collaborative Piano and Body Mapping with the objective of imparting fundamental collaborative piano skills integrated with proper body use. The author hopes to inform educators of the benefits of prioritizing health among their students and to provide a Body Mapping foundation upon which their students can build technique.
ContributorsBindel, Jennifer (Author) / Campbell, Andrew (Thesis advisor) / Doan, Jerry (Committee member) / Rogers, Rodney (Committee member) / Ryan, Russell (Committee member) / Schuring, Martin (Committee member) / Arizona State University (Publisher)
Created2013
Description
The purpose of the paper is to outline the process that was used to write a reduction for Henry Brant's Concerto for Alto Saxophone and Orchestra, to describe the improvements in saxophone playing since the premiere of the piece, and to demonstrate the necessity of having a reduction in the process of learning a concerto. The Concerto was inspired by internationally known saxophonist, Sigurd Rascher, who demonstrated for Brant the extent of his abilities on the saxophone. These abilities included use of four-octave range and two types of extended techniques: slap-tonguing and flutter-tonguing. Brant incorporated all three elements in his Concerto, and believed that only Rascher had the command over the saxophone needed to perform the piece. To prevent the possibility of an unsuccessful performance, Brant chose to make the piece unavailable to saxophonists by leaving the Concerto without a reduction. Subsequently, there were no performances of this piece between 1953 and 2001. In 2011, the two directors of Brant's Estate decided to allow for a reduction to be written for the piece so that it would become more widely available to saxophonists.
ContributorsAmes, Elizabeth (Pianist) (Author) / Ryan, Russell (Thesis advisor) / Levy, Benjamin (Committee member) / Hill, Gary (Committee member) / Campbell, Andrew (Committee member) / Arizona State University (Publisher)
Created2013
Description
Single molecule DNA Sequencing technology has been a hot research topic in the recent decades because it holds the promise to sequence a human genome in a fast and affordable way, which will eventually make personalized medicine possible. Single molecule differentiation and DNA translocation control are the two main challenges in all single molecule DNA sequencing methods. In this thesis, I will first introduce DNA sequencing technology development and its application, and then explain the performance and limitation of prior art in detail. Following that, I will show a single molecule DNA base differentiation result obtained in recognition tunneling experiments. Furthermore, I will explain the assembly of a nanofluidic platform for single strand DNA translocation, which holds the promised to be integrated into a single molecule DNA sequencing instrument for DNA translocation control. Taken together, my dissertation research demonstrated the potential of using recognition tunneling techniques to serve as a general readout system for single molecule DNA sequencing application.
ContributorsLiu, Hao (Author) / Lindsay, Stuart M (Committee member) / Yan, Hao (Committee member) / Levitus, Marcia (Committee member) / Arizona State University (Publisher)
Created2013
Discovering Puerto Rican art song: a research project on four art song works by Héctor Campos Parsi
Description
Puerto Rico has produced many important composers who have contributed to the musical culture of the nation during the last 200 years. However, a considerable amount of their music has proven to be difficult to access and may contain numerous errors. This research project intends to contribute to the accessibility of such music and to encourage similar studies of Puerto Rican music. This study focuses on the music of Héctor Campos Parsi (1922-1998), one of the most prominent composers of the 20th century in Puerto Rico. After an overview of the historical background of music on the island and the biography of the composer, four works from his art song repertoire are given for detailed examination. A product of this study is the first corrected edition of his cycles Canciones de Cielo y Agua, Tres Poemas de Corretjer, Los Paréntesis, and the song Majestad Negra. These compositions date from 1947 to 1959, and reflect both the European and nationalistic writing styles of the composer during this time. Data for these corrections have been obtained from the composer's manuscripts, published and unpublished editions, and published recordings. The corrected scores are ready for publication and a compact disc of this repertoire, performed by soprano Melliangee Pérez and the author, has been recorded to bring to life these revisions. Despite the best intentions of the author, the various copyright issues have yet to be resolved. It is hoped that this document will provide the foundation for a resolution and that these important works will be available for public performance and study in the near future.
ContributorsRodríguez Morales, Luis F., 1980- (Author) / Campbell, Andrew (Thesis advisor) / Buck, Elizabeth (Committee member) / Holbrook, Amy (Committee member) / Kopta, Anne (Committee member) / Ryan, Russell (Committee member) / Arizona State University (Publisher)
Created2013
Description
Artistic trends of the mid-nineteenth century demonstrate the popularity of incorporating Asian elements into various artistic media. This paper discusses why the stereotypical Asian female provided an attractive character for operatic librettists, composers and audiences. To support the discussion, six operas from 1885 to 2010 are examined, and the dramatic and musical portrayal of representative female characters is discussed. The familiar character of Cio-cio-san from Giocamo Puccini's Madama Butterfly (1904) provides a foundation to discuss these stereotypical Asian female characteristics, specifically one archetype, that of the naïve, yet sexually desirable female. Prior to Cio-cio-san, Sir W. S. Gilbert and Sir Arthur Sullivan's Yum-Yum from The Mikado (1885), Iris of Pietro Mascagni's Iris (1898) exemplify this archetype, as does Liù from Puccini's Turandot (1924). At the other extreme is the icy, cold and bloodthirsty archetype found in the title role of Puccini's Turandot and Katisha from The Mikado. Chiang Ch'ing (also known as Madame Mao) from John Adams's Nixon in China (1987), and Madame White Snake from Chinese-American composer Zhou Long's Madame White Snake (2010) feature leading characters that demonstrate elements of both of these archetypes, and this combination of the two archetypes yields more complex and richer characters. These two extremes of the female Asian stereotype and the evolution of these characteristics provide an interesting outlook on the incorporation of non-Western musical styles into these operas, and the understanding of a Western perception of foreign peoples, especially foreign females.
ContributorsLo, Wan-Yi (Author) / Campbell, Andrew (Thesis advisor) / Carpenter, Ellon (Committee member) / Kopta, Anne (Committee member) / Mills, Robert (Committee member) / Ryan, Russell (Committee member) / Arizona State University (Publisher)
Created2013