DRUM - Digital Repository at the University of Maryland
DRUM collects, preserves, and provides public access to the scholarly output of the university. Faculty and researchers can upload research products for rapid dissemination, global visibility and impact, and long-term preservation.
Submit to DRUM
To submit an item to DRUM, login using your UMD credentials. Then select the "Submit Item to DRUM" link in the navigation bar. View DRUM policies and submission guidelines.
Equitable Access Policy
The University of Maryland Equitable Access Policy provides equitable, open access to the University's research and scholarship. Faculty can learn more about what is covered by the policy and how to deposit on the policy website.
Theses and Dissertations
DRUM includes all UMD theses and dissertations from 2003 forward.
List of Communities
Collections Organized by Department
UM Community-managed Collections
Recent Submissions
Cloning and Expression of Human RPS24 into E.coli and the HEK293 Cell Line
(2024-04-26) Merrifield, Katherine; Apgar, Sofia; Jessica Whitney; Argueta, Vicky; Zeidan, Quira
Ribosome biogenesis is the process of constructing ribosomes and requires ribosomal RNA, ribosomal proteins (RPs), and assembly factors. The products of eukaryotic ribosome biogenesis are the large 60S subunit and the small 40S subunit, of which RPS24 is crucial in its formation. In addition to its translational roles, RPS24 is associated with regulating cell growth and proliferation, apoptosis, and DNA damage response. We hypothesize that the extra-ribosomal functions of RPS24 are impacted by its post-translational modifications (PTMs). To elucidate the functions of these PTMs, the coding sequence of human RPS24 was cloned into pNH-TrxT and pNIC28-Bsa4 bacterial expression vectors via ligase-independent cloning. The recombinant plasmids were then transformed into BL21 E.coli cells, and initial trials were conducted to optimize growth and expression conditions for the two transformed strains. Protein expression was determined using SDS-PAGE and Coomassie staining, the results of which indicated moderate levels of the RPS24 fusion protein in cells transformed with both the recombinant plasmids. RPS24 expression was observed without IPTG induction, indicating leaky expression. In parallel experiments, we investigated the overexpression of RPS24 in HEK293 cells from the plasmid pcDNA3.1(C)DYK, and successful transfection was confirmed by SDS-PAGE and Western Blot analysis. We aim to investigate the role of RPS24 PTMs in cell proliferation and viability under various stress conditions to evaluate their impact on tumor development.
Expressing Human Ribosomal Proteins S20 and S26 in Modified E.coli (BL21) Cells
(2024-04-26) Lee, Maia; Quenano, Erika; Owen, Jennifer; Ngouba, Andrea; Zahniser, Hannah; Tafesh, Talah; Ofoegbu, Stacie; Weber, Louis; Quira Zeidan
Ribosomes are fundamental in creating proteins that contribute to the proper functioning of cellular processes. Thus, ribosome biogenesis must occur with integrity by appropriately expressing ribosomal proteins (RPs). Mutations in the genes of human RPS20 and RPS26 have been linked to disruption in the translation machinery and anomalies in the production of red blood cells, leading to diseases such as Diamond-Blackfan Anemia. To develop our understanding of the role of mutations in human RPS20 and RPS26, we performed ligase-independent cloning of their coding sequence into pNIC28-Bsa4 and pNH-TrxT and transformed these plasmids into a BL21 E. coli expression system. IPTG induction was used to determine conditions necessary to maximize the expression of soluble RPS20 and RPS26. After SDS-PAGE, Coomassie staining showed successful expression of RPS26 in response to IPTG induction, whereas RPS20 was produced regardless of IPTG presence. Human RPs expressed in bacteria are often purified from insoluble inclusion bodies; thus, determining the conditions for maximal solubility provides an optimal strategy for studying fusion proteins. Future research on the effects of the overexpression of RPS20 and RPS26 on human cell growth and viability will expand our understanding of the complex mechanisms of human ribosome biogenesis and its role in disease.
Stable accumulation of human ribosomal proteins SA, S2, S3, S4X, S6, S7 in rabbit reticulocyte cell-free expression system
(2024) Neuheisel, Mary Louisa; Streit, Julia; Zeidan, Quira
Mutations in human ribosomal proteins (RPs) contribute to severe, tissue-specific pathologies, indicating roles that extend beyond protein translation. RPSA is associated with pancreatic cell migration and congenital asplenia, while RPS3, RPS4X, and RPS7 are linked to Diamond-Blackfan anemia (DBA). While we understand the nucleolar assembly of RPs, the cytoplasmic events preceding RP translocation into the nucleus remain unclear. This work investigated the expression and posttranslational modifications (PTMs) of human RPSA, RPS2, RPS3, RPS4X, RPS6, and RPS7 using Rabbit Reticulocyte Lysate (RRL), an enucleated cell-free system that allows the study of cytosolic modifications crucial for RP stability. We conducted coupled transcription/translation experiments with Western blot analyses and immunoprecipitations with anti-FLAG antibodies and magnetic beads to analyze RP expression and cytosolic PTMs. Our results demonstrated the robust and stable expression of RPs, with distinct abundance and stability among different proteins despite expression from the same promoter under identical conditions. RPSA, RPS3, and RPS6 exhibited a molecular weight of approximately 8-10 kDa higher than predicted, suggesting PTM with a peptide. We also detected the presence of enzymes for O-GlcNAc modification within the RRL system. Treatment with TMG, an O-GlcNAcase inhibitor, resulted in increased O-GlcNAcylation levels. We propose that the RRL system is an invaluable tool for probing the roles of PTMs in the extraribosomal activity of RPs within the cytoplasm, representing a relevant and accessible model to deepen our understanding of ribosomopathies and their future therapeutic interventions.
PERFORMANCE OF THE VIOLIN CONCERTO AND SONATAS OF JOHANNES BRAHMS WITH AN ANALYSIS OF JOSEPH JOACHIM'S INFLUENCE ON HIS VIOLIN CONCERTO
(1997) Hsieh, I-Chun; Heifetz, Daniel; Music; Digital Repository at the University of Maryland; University of Maryland (College Park, MD)
This dissertation consists of a performance project and
extensive studies of selected works by Johannes Brahms, including
the Violin Concerto, Sonatensatz, and three Violin Sonatas.
The performance project was presented in two recitals at the
University of Maryland, College Park, on November 14, 1997, and
November 16, 1997. The first recital featured Brahms' s Sonatensatz in C Minor, Violin Sonata No. I, Op. 78 in G Major, and Violin
Sonata No. 3, Op. 108, in D Minor. The second recital included
Brahms' s Violin Sonata No.2, Op. 100, in A Major and Violin
Concerto Op. 77, in D Major. Section One gives an overview of this dissertation project.
Section Two introduces the violin virtuoso Joseph Joachim, his
relationship with Johannes Brahms, and Brahms' s life and major
violin works. This section also analyzes Joachim' s performance
practice and his teaching style. The end of this section focuses on the
influence of Joseph Joachim on Brahms' s Violin Concerto and
indicates the differences between Brahms' s original manuscript and
the version suggested by Joachim.
Section Three is composed of the programs of the two recitals.
Section Four consists of program notes for the two recitals.
The first recital was performed by I-Chun Hsieh, violin and
Roy Hakes, piano. The second recital was performed by I-Chun
Hsieh, violin and Chia-Hsuan Lee, piano.
Baroque Plague Imagery and Tridentine Church Reforms
(1990) Boeckl, Christine M.; Pressly, William; Art History & Archaeology; Digital Repository at the University of Maryland; University of Maryland (College Park, MD)
This dissertation aims to achieve two goals: one, to assemble
as many facts as possible about the plague, regardless of period, and
to relate this material to images; and two, to present a well-defined
group of religious baroque plague paintings in the context of social,
political and religious history. This inquiry is primarily concerned
with scenes that portray saints actively involved in charitable
pursuits, dispensing the sacraments to victims of the most dreaded
disease, the bubonic plague.
Chapter I contains a bibliographical essay, divided into three
parts: medicine, theology, and art history. The next chapter considers
the sources and the formation of baroque plague iconography. The
remaining two chapters discuss "documentary" plague scenes and
how they relate to historic events. They are presented in two
sections: Italy and transalpine countries.
This interdisciplinary research resulted in a number of
observations. First, these narrative plague scenes were produced in
Italy and in Catholic countries bordering Protestant regions:
Switzerland, France, Flanders, and in the Habsburg Empire (excluding
Spain). Second, the painters were mostly Italian or Italian-trained.
Third, the artists observed not only the requirements specified by the
Church in the 1563 Tridentine Decree on the Arts but also reflected
in their work the catechetic teachings of the Council. Fourth, these
religious scenes were not votive paintings but doctrinal images that
served either didactic or polemic functions. Fifth, the scenes were
not intended as memento mori; rather, the iconology conveyed
positive images which emphasized that the faithful needed the
Roman Catholic clergy to gain life-everlasting. Sixth, these plague
paintings were important documents not as recordings of the
conditions experienced during an epidemic but as historic testimony
of liturgical practices. Last, these selected scenes mirrored the
baroque Church's views on the ultimate questions about life and
death.