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- Genre: Doctoral Dissertation
- Creators: Rogers, Rodney
- 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
Several contemporary clarinet works use Chinese folk music elements from different regions in new compositions to entice listener's and performer's appreciation of Chinese culture. However, to date, limited academic research on this topic exists. This research paper introduces six contemporary clarinet works by six Chinese composers: Qigang Chen's Morning Song, Yan Wang's Mu ma zhi ge (The Song of Grazing Horses), An-lun Huang's Capriccio for Clarinet and Strings Op. 41, Bijing Hu's The Sound of Pamir Clarinet Concerto, Mei-Mi Lan's Concerto for Clarinet and String Orchestra with Harp and Percussion, and Yu-Hui Chang's Three Fantasias for Solo Clarinet in B-flat. They are examined from different perspectives, including general structure, style, and rejuvenated folk music use. The focus of this research paper is to investigate the use of Chinese folk music in several works in collaboration with the composers. The author found that although contemporary composers use Chinese folk music differently in their works (i.e., some use melodies, others use harmony, while others use modes), each work celebrates the music and culture of the folk music on which the pieces are based. It is the author's hope to stimulate people's interest in music using Chinese folk music elements, and bring these lesser known works into the common clarinet repertoire.
ContributorsFeng, Chiao-Ting (Author) / Spring, Robert (Thesis advisor) / Gardner, Joshua (Committee member) / Micklich, Albie (Committee member) / Rogers, Rodney (Committee member) / Schuring, Martin (Committee member) / Arizona State University (Publisher)
Created2013
Description
Bright Summer, a one-movement piece for orchestra, was composed in Arizona, and completed in February 2013. The piece is approximately twelve minutes long. The motivation for writing this piece was the death of my mother the year before, in 2012. The prevailing mood of this work is bright and pleasant, expressing my mother's cheerful personality when she was alive. It also portrays bright summer days which resemble my mother's spirit. Thus, soundscape plays an important role in this work. It depicts summer breeze, rustling sounds of leaves, and, to translate a Korean saying, "high blue skies." This soundscape opens the piece as well as closes it. In the middle section, the fast upbeat themes represent my mother's witty and optimistic personality. The piece also contains the presence of a hymn tune, The Love of God is Greater Far, which informs the motivic content and also functions as the climax of the piece. It was my mother's favorite hymn and we used to sing it together following her conversion to Christianity. The piece contains three main sections, which are held together by transitional material based on the soundscape and metric modulations. Unlike my earlier works, Bright Summer is tonal, with upper tertian harmonies prevailing throughout the piece. However, the opening and closing soundscapes do not have functional harmonies. For example, tertian chords appear and vanish silently, leaving behind some resonant sounds without any harmonic progression. Overall, the whole piece is reminiscent of my mother who lived a beautiful life.
ContributorsKim, JeeYeon (Composer) / DeMars, James (Thesis advisor) / Hackbarth, Glenn (Committee member) / Rogers, Rodney (Committee member) / Levy, Benjamin (Committee member) / Rockmaker, Jody (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
There are a significant number of musical compositions for violin by composers who used folk songs and dances of various cultures in their music, including works by George Enescu, Béla Bartók and György Ligeti. Less known are pieces that draw on the plethora of melodies and rhythms from Turkey. The purpose of this paper is to help performers become more familiar with two such compositions: Fazil Say's Sonata for Violin and Piano and Cleopatra for Solo Violin. Fazil Say (b. 1970) is considered to be a significant, contemporary Turkish composer. Both of the works discussed in this document simulate traditional "Eastern" instruments, such as the kemenҫe, the baðlama, the kanun and the ud. Additionally, both pieces use themes from folk melodies of Turkey, Turkish dance rhythms and Arabian scales, all framed within traditional structural techniques, such as ostinato bass and the fughetta. Both the Sonata for Violin and Piano and Cleopatra are enormously expressive and musically interesting works, demanding virtuosity and a wide technical range. Although this document does not purport to be a full theoretical analysis, by providing biographical information, analytical descriptions, notes regarding interpretation, and suggestions to assist performers in overcoming technical obstacles, the writer hopes to inspire other violinists to consider learning and performing these works.
ContributorsKalantzi, Panagiota (Author) / Jiang, Danwen (Thesis advisor) / Hill, Gary (Committee member) / Rogers, Rodney (Committee member) / Rotaru, Catalin (Committee member) / Arizona State University (Publisher)
Created2013
Description
The purpose of this project is twofold: to contribute to the literature of chamber ensembles comprising mixed wind, string, and percussion instruments by producing arrangements of three piano rags by William Bolcom; and to highlight Bolcom's pivotal role in the ragtime revival of the 1960's and 1970's. Through his influence as a scholar, composer, and performer, Bolcom (b. 1938), one of the most prominent American composers of his generation, helped garner respect for ragtime as art music and as one of America's great popular music genres. Bolcom's 3 Ghost Rags were written in the tradition of classic piano rags, but with a compositional sensibility that is influenced by the fifty years that separate them from the close of the original ragtime era. The basis for the present orchestrations of 3 Ghost Rags is the collection of instrumental arrangements of piano rags published by Stark Publishing Co., entitled Standard High-Class Rags. More familiarly known as the "Red Back Book," this publication was representative of the exchange of repertoire between piano and ensembles and served as a repertory for the various ragtime revivals that occurred later in the twentieth century. In creating these orchestrations of Bolcom's piano rags, the author strove to provide another medium in which Bolcom's music could be performed, while orchestrating the music for an historically appropriate ensemble.
ContributorsMelley, Eric Charles (Author) / Hill, Gary W. (Thesis advisor) / Bailey, Wayne (Committee member) / Norton, Kay (Committee member) / Rogers, Rodney (Committee member) / Russell, Timothy (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