Matching Items (177)
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
This final research paper provides both a performer's perspective and a recording of double clarinet literature by William O. Smith (b. 1926), Eric Mandat (b. 1957), and Jody Rockmaker (b. 1961). The document includes musical examples, references to the recording, and interviews with the composers. The first chapter contains a brief literature review of sources on world double clarinets, biographies of the above-mentioned composers, and other pertinent information. Chapters 2-4 include the performer's perspective on the following works: Epitaphs for Double Clarinet by William O. Smith, Double Life for Solo Clarinet by Eric Mandat, and two compositions by Jody Rockmaker, Half and Half for demi-clarinet in A, and Double Dip. The final chapter examines how double clarinet music has evolved, the challenges and limitations of the repertoire, and the future of the double clarinet genre.
ContributorsEndel, Kimberly Michelle (Author) / Spring, Robert S (Thesis advisor) / Gardner, Joshua (Committee member) / Norton, Kay (Committee member) / Micklich, Albie (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
About piano students who display disruptive behavior and perform far below reasonable expectations, teachers first conclude that they are lazy, rude, disinterested, and/or lacking intelligence or ability. Most dismiss such students from studios and advise parents to discontinue lessons. In truth, many of these students are both highly gifted and also have a learning disability. Examined literature shows that the incidence of dyslexia and other learning disabilities in the gifted learner population is several times that of the regular learner population. Although large volumes of research have been devoted to dyslexia, and more recently to dyslexia and music (in the classroom and some in individual instrumental instruction), there is no evidence of the same investigation in relation to the specific needs of highly gifted dyslexic students in learning to play the piano. This project examines characteristics of giftedness and dyslexia, gifted learners with learning disabilities, and the difficulties they encounter in learning to read music and play keyboard instruments. It includes historical summaries of author's experience with such students and description of their progress and success. They reveal some of practical strategies that evolved through several decades of teaching regular and gifted dyslexic students that helped them overcome the challenges and learn to play the piano. Informal conversations and experience exchanges with colleagues, as well as a recently completed pilot study also showed that most piano pedagogues had no formal opportunity to learn about this issue and to be empowered to teach these very special students. The author's hope is to offer personal insights, survey of current knowledge, and practical suggestions that will not only assist piano instructors to successfully teach highly gifted learners with dyslexia, but also inspire them to learn more about the topic.
ContributorsVladikovic, Jelena (Author) / Humphreys, Jere T. (Thesis advisor) / Meir, Baruch (Thesis advisor) / Norton, Kay (Committee member) / Hamilton, Robert (Committee member) / Arizona State University (Publisher)
Created2013
Description
The end of the nineteenth century was an exhilarating and revolutionary era for the flute. This period is the Second Golden Age of the flute, when players and teachers associated with the Paris Conservatory developed what would be considered the birth of the modern flute school. In addition, the founding in 1871 of the Société Nationale de Musique by Camille Saint-Saëns (1835-1921) and Romain Bussine (1830-1899) made possible the promotion of contemporary French composers. The founding of the Société des Instruments à Vent by Paul Taffanel (1844-1908) in 1879 also invigorated a new era of chamber music for wind instruments. Within this groundbreaking environment, Mélanie Hélène Bonis (pen name Mel Bonis) entered the Paris Conservatory in 1876, under the tutelage of César Franck (1822-1890). Many flutists are dismayed by the scarcity of repertoire for the instrument in the Romantic and post-Romantic traditions; they make up for this absence by borrowing the violin sonatas of Gabriel Fauré (1845-1924) and Franck. The flute and piano works of Mel Bonis help to fill this void with music composed originally for flute. Bonis was a prolific composer with over 300 works to her credit, but her works for flute and piano have not been researched or professionally recorded in the United States before the present study. Although virtually unknown today in the American flute community, Bonis's music received much acclaim from her contemporaries and deserves a prominent place in the flutist's repertoire. After a brief biographical introduction, this document examines Mel Bonis's musical style and describes in detail her six works for flute and piano while also offering performance suggestions.
ContributorsDaum, Jenna Elyse (Author) / Buck, Elizabeth (Thesis advisor) / Holbrook, Amy (Committee member) / Micklich, Albie (Committee member) / Schuring, Martin (Committee member) / Norton, Kay (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