Molecular Genetics Commons^™

Epigenetics Of Host-Pathogen Interactions: The Effect Of Acetosyringone On Ti Plasmid Methylation Patterns In Agrobacterium Tumefaciens C58, Genevieve Roberts

Honors Projects

The plant pathogen Agrobacterium tumefaciens C58 can transfer a portion of its tumor-inducing (Ti) plasmid to plant hosts in response to the plant wound signals, including Acetosyringone. The portion transferred is aptly titled Transfer DNA (T-DNA) which encode genes involved in tumor production and biosynthesis of a unique bacterial food source called opines that provide an advantage to the inducing Agrobacterium. The Ti plasmid contains a number of genes, including the virulence region that enables T-DNA transfer. Epigenetics investigates how chemical modifications to DNA that don’t alter sequence are used to control gene expression (for example, genes involved in pathogen …

Body Size Regulation Via Bmp Signaling In Caenorhabditis Elegans, Uday Madaan

Dissertations, Theses, and Capstone Projects

The body size of an organism can be a crucial determinant of access to nutrition, reproductive success and overall survival in the wild. However, how body size of an individual is determined is incompletely understood. Body size is a complex trait determined by multiple pathways and genes, making it difficult to understand the role of individual genes and pathways in determining overall size. In Caenorhabditis elegans, a homolog of Bone Morphogenetic Proteins (BMP) is a major regulator of body size; functional loss of DBL-1 leads to a small body size. Due to a drastic change in body size in dbl-1 …

The Population Genetics Of Morro Bay Eelgrass (Zostera Marina), Julia Gardner Harencar

Master's Theses

Seagrass populations are in decline worldwide. Zostera marina (eelgrass), one of California’s native seagrasses, is no exception to this trend. In the last 8 years, Morro Bay, California has lost 95% of its eelgrass. Eelgrass is an ecosystem engineer, providing important ecosystem services such as sediment stabilization, nutrient cycling, and nursery habitats for fish. The failure of recent restoration efforts necessitates a better understanding of the causes of eelgrass decline in this estuary. Previous research on eelgrass in California has demonstrated a link between population genetic diversity and eelgrass bed health, ecosystem functioning, and resilience to disturbance and extreme climatic …

Zinc Transporters Ybtx And Znuabc Are Required For The Virulence Of Yersinia Pestis In Bubonic And Pneumonic Plague In Mice, Alexander G. Bobrov, Olga Kirillina, Marina Y. Fosso, Jacqueline D. Fetherston, M. Clarke Miller, Tiva T. Vancleave, Joseph A. Burlison, William K. Arnold, Matthew B. Lawrenz, Sylvie Garneau-Tsodikova, Robert D. Perry

Microbiology, Immunology, and Molecular Genetics Faculty Publications

A number of bacterial pathogens require the ZnuABC Zinc (Zn²⁺) transporter and/or a second Zn²⁺ transport system to overcome Zn²⁺ sequestration by mammalian hosts. Previously we have shown that in addition to ZnuABC, Yersinia pestis possesses a second Zn²⁺ transporter that involves components of the yersiniabactin (Ybt), siderophore-dependent iron transport system. Synthesis of the Ybt siderophore and YbtX, a member of the major facilitator superfamily, are both critical components of the second Zn²⁺ transport system. Here we demonstrate that a ybtX znu double mutant is essentially avirulent in mouse models of bubonic and pneumonic …

The 'Pseudomonas Aeruginosa' Psl Polysaccharide Is A Social But Noncheatable Trait In Biofilms, Yasuhiko Irie, Aled E. Roberts, Kasper N. Kragh, Vernita D. Gordon, Jaime B. Hutchison, Rosalind J. Allen, Gavin Melaugh, Thomas Bjarnsholt, Stuart A. West, Stephen P. Diggle

Biology Faculty Publications

Extracellular polysaccharides are compounds secreted by microorganisms into the surrounding environment, and they are important for surface attachment and maintaining structural integrity within biofilms. The social nature of many extracellular polysaccharides remains unclear, and it has been suggested that they could function as either cooperative public goods or as traits that provide a competitive advantage. Here, we empirically tested the cooperative nature of the PSL polysaccharide, which is crucial for the formation of biofilms in Pseudomonas aeruginosa. We show that (i) PSL is not metabolically costly to produce; (ii) PSL provides populationlevel benefits in biofilms, for both growth and antibiotic …

The Santa Ana Speckled Dace (Rhinichthys Osculus): Phylogeography And Molecular Evolution Of The Mitochondrial Dna Control Region, James Jay Vanmeter

Electronic Theses, Projects, and Dissertations

The purpose of this genetic study of the Santa Ana Speckled Dace Rhinichthys osculus was three-fold. The first goal was to characterize the molecular structure of the mtDNA control region of R. osculus. An 1143 base-pair region of the mitochondrial DNA genome, which included the complete control region was sequenced for all individuals. Analysis of the sequence data revealed that the molecular structure of the speckled dace control region was similar to the molecular structure described for other vertebrate taxa. The speckled dace control region contains three major domains, which vary in base frequency as well as in the frequency …

Dna Sequences Of The Mitochondrial Cytochrome C Oxidase I (Coi) Genes From Deep Sea Fishes. Cruises Dp01 And Dp02 From May 2015 - August 2015, Andrea Bernard, Max Weber, Kimberly A. Finnegan, Mahmood S. Shivji, Ron Eytan

DEEPEND Datasets

The deep sea ecosystem is believed to contain the highest biomass of fish in the oceans. However, the taxonomic diversity in this ecosystem is incompletely described and likely to be vastly underestimated. DNA sequence data (barcodes) have become a key tool to discover hidden biodiversity. We generated mitochondrial DNA barcode datasets based on the Cytochrome c oxidase I (COI) gene from deep sea fishes. These data were analyzed using phylogenetic and statistical methods to reveal cryptic species and make taxonomic linkages between adult fishes and their early life stages. These datasets were generated from fishes collected in the Northern Gulf …

Does Genotype Correlate With Phenotype? Evaluating Ruffed Lemur (Varecia Spp.) Color Vision Using Subject Mediated Automatic Remote Testing Apparatus (Smarta), Raymond Vagell

Theses and Dissertations

Ruffed lemur (Varecia spp.) color vision research was conducted using a multidisciplinary approach: psychophysics, genetic analysis, technology, and animal training. The behavioral manifestation of Varecia spp. trichromacy was shown using a touchscreen apparatus (SMARTA). Trichromats performed better than dichromats when discriminating red from green (G₂ = 78.10, p < 0.001).

Chromatin-Modifying Agents Convert Fibroblasts To Oct4+ And Vegfr-2+ Capillary Tube-Forming Cells, Anita Wary, Neil Wary '18, Jugajyoti Baruah, Victoria Mastej, Kishore K. Wary

Student Publications & Research

Rationale
The human epigenome is plastic. The goal of this study was to address if fibroblast cells can be epigenetically modified to promote neovessel formation.

Methods and results
Here, we used highly abundant human adult dermal fibroblast cells (hADFCs) that were treated with the chromatin-modifying agents 5-aza-2'-deoxycytidine and trichostatin A, and subsequently subjected to differentiation by activating Wnt signaling. Our results show that these epigenetically modified hADFCs increasingly expressed β-catenin, pluripotency factor octamer-binding transcription factor-4 (OCT4, also known as POU5F1), and endothelial cell (EC) marker called vascular endothelial growth factor receptor-2 (VEGFR-2, also known as Fetal Liver Kinase-1). In microscopic …

Phenotypic Variation And Genetic Purity Of The Original 'Prime-Jim®' X 'Arapaho' Population, Loren M. Luther

Horticulture Undergraduate Honors Theses

In 2013, Castro et al., produced the first linkage map for primocane-fruiting blackberries using the cross ‘Prime-Jim®’ x ‘Arapaho.’ This mapping population has been maintained since the original experiment with the hopes of conducting future studies. Further research was determined to be needed on the population to better characterize additional phenotypic traits, and since nine years had passed since the seedlings were established, there was a need to assess the continued genetic purity of the population using molecular methods. Phenotypic data was collected to analyze the variation of the population including soluble solids content, titratable acidity, average weight, shape, and …

Characterization Of E-Cadherin Regulation In Response To Zeb1 Inhibition In Endometrial Cancer Cell Lines, Chidozie Paul Chukwu

Graduate School of Biomedical Sciences Theses and Dissertations

Epithelial to mesenchymal transition (EMT) is the process in which cells lose their epithelial structure during gastrulation. This process also affects the migration and movement of tumor cells and promotes invasion and metastases of endometrial carcinomas. Down-regulation of E-cadherin (CDH1) by transcription factors is the key target of EMT modulators and is achieved mainly by ZEB1 (zinc finger E-box binding homeobox 1). Current research looking at restoration of E-cadherin expression in vitro involves the use of small molecules such as histone deacetylase (HDAC) inhibitors and DNA methyltransferase inhibitors. Trichostatin A (TSA) and small interfering ribonucleic acid (siRNA) are tools that …

The Role Of The Expansion Segment 7 Of 25s Rrna During Oxidative Stress In Saccharomyces Cerevisiae, Ethan Gardner

Graduate School of Biomedical Sciences Theses and Dissertations

Translation is an essential process for protein expression in both eukaryotes and prokaryotes. Like any cellular process, translational factors are prone to damage when the cell is under stress. One common stressor that nearly all cells may experience is abnormal levels of reactive oxygen species (ROS). Damage caused by ROS has been associated with disease ranging from neurodegenerative impairments, to the aging process of cells. These oxygen radicals are capable of damaging a litany of molecules including nucleic acids, and molecular factors involved in translation. It has been shown that tRNA can be cleaved upon ROS-induced stress and these fragments …

Modeling 3d Retinogenesis In Mouse Embryonic Stem Cells Following Crispr-Mediated Crx Knockdown, Pooja Prasad

Dissertations, Masters Theses, Capstones, and Culminating Projects

An emerging technology known as three-dimensional (3D) tissue engineering has allowed scientists to mimic tissues found in vivo. Previous studies indicate that it is possible to differentiate dissociated mouse embryonic stem cells (mESCs) into 3D retinal tissues in vitro (Bertacchi, 2015; Eiraku, 2012). The newly differentiated retinal tissues are said to encompass all of the major components found in retinal tissues. The generation of in vitro 3D tissues holds great potential in terms of patient-specific disease modeling. Although various diseases have been well-studied in animal models, there are limitations with regards to patient-specificity. The generation of animal models to study …

Science, Between The Lines: Rosalind Franklin, Rachael Renzi

Senior Honors Projects

The purpose of this paper is to contrast scientific rhetoric with creative biographical writing style. Both forms of communication rely on specific syntax and styles to describe research findings to the appropriate audiences, but the often passive format of scientific literature excludes the passionate reasonings of the researcher. In order to understand why the scientist is kept separate from his or her published findings, I am going to experiment with the two contrasting forms. Throughout the paper, each form will be tested and used at varying levels. Both scientific and creative writing will be mixed. To emphasize the technically dense …

Targeted Germ Cell Knockdown Of Xlr3 In Mus Musculus, Caity Miller

Honors Scholar Theses

The X chromosome is rather distinct from autosomes due to the unique regulatory and functional characteristics it exhibits. Xlr3 is just one gene in a superfamily of highly related, and homologous genes found on the X chromosome in mice. Xlr3 is part of a complex, imprinted locus, of which the function is not well understood, although its protein product can be found in testis and oocyte. However, it is theorized that the region may be implicated in the progression of meiosis, due to localization of XLR3. This study sought to characterize the function of Xlr3 through the use of a …

Proteomic Identification Of Histone Post-Translational Modifications Induced By Dna Double-Strand Breaks And Novel Proteins Involved In The Dna Damage Response, Pingping Wang

Dissertations and Theses (Open Access)

Inaccurate repair of DNA double-strand breaks (DSBs) can lead to DNA mutation and chromosome rearrangements, causing human diseases such as cancer. Although we know the basic mechanisms of DSB repair, the added complexities in the chromatin context are unclear. This is partially due to the lack of unbiased systems for identifying proteins and post-translational modifications (PTMs) involved in DSB repair. In this work, we established a novel method, termed DSB-ChAP-MS (Double Strand Break-Chromatin Affinity Purification with Mass Spectrometry), for the affinity purification of a sequence-specific single copy endogenous chromosomal locus containing a DSB, followed by the proteomic identification of enriched …

Analysis Of The Biochemical And Cellular Activities Of Substrate Binding By The Molecular Chaperone Hsp110/Sse1, Veronica M. Garcia

Dissertations and Theses (Open Access)

Molecular chaperones ensure protein quality during protein synthesis, delivery, damage repair, and degradation. The ubiquitous and highly conserved molecular chaperone 70-kDa heat-shock proteins (Hsp70s) are essential in maintaining protein homeostasis by cycling through high and low affinity binding of unfolded protein clients to facilitate folding. The Hsp110 class of chaperones are divergent relatives of Hsp70 that are extremely effective in preventing protein aggregation but lack the hallmark folding activity seen in Hsp70s. Hsp110s serve as Hsp70 nucleotide exchange factors (NEF) that facilitate the Hsp70 folding cycle by inducing release of protein substrate from Hsp70, thus recycling the chaperone for a …

Understanding The Mechanism Of Genomic Instability During Replicative Aging In Budding Yeast, Sangita Pal

Dissertations and Theses (Open Access)

Aging brings a gradual decline in molecular fidelity and biological functionality, resulting in age related phenotypes and diseases. Despite continued efforts to uncover the conserved aging pathways among eukaryotes, exact molecular causes of aging are still poorly understood. One of the most important hallmarks of aging is increased genomic instability. However, there remains much ambiguity as to the cause. I am studying the replicative life span (RLS) of the genetically tractable model organism Saccharomyces cerevisiae, or budding yeast using the innovative “mother enrichment program” as the method to isolate unparalleled numbers of aged yeast cells to investigate the molecular changes …

A Genetic Analysis Of Nuclear Functions Of The Lipin Protein In Drosophila Melanogaster, Xeniya Rudolf

Graduate Theses and Dissertations

Lipins are a family of proteins that have critical functions in the control of fat storage and energy homeostasis. Biochemically, lipins have two functions. They provide an enzymatic activity (phosphatidate phosphatase or PAP activity) in the glycerol-3 phosphate pathway that leads to the production of storage fats (triacylglycerols). In addition, they play a role in the regulation of genes in the cell nucleus as transcriptional co-regulators. The PAP activity of lipins has been widely studied in a number of organisms. However, the transcriptional co-regulator function is not as well described in the literature. The transcriptional function of lipins depends on …

Genetic Polymorphisms Of The Glucocorticoid Receptor And Interleukin-8 Receptor Genes Are Related To Production Traits And Hair Coat Score In Crossbred Cattle, Avery Deaton

Animal Science Undergraduate Honors Theses

The objective of this thesis was to identify polymorphisms in the glucocorticoid receptor (GR) and interleukin-8 receptor (CXCR2) genes and to associate genotypes between the above mentioned polymorphisms and production traits in crossbred cattle. The hypothesis was that polymorphisms will exist for GR and CXCR2 genes and will be linked to production traits. Glucocorticoid receptors have been positively associated with higher milk yields, lactose content, feed intake, and feed conversion rates. Interleukin-8 genes are part of the innate immune response and help with many aspects of female reproduction health, such as protecting the embryo from the maternal immune system during …

The Effect Of Galactose On The Expression Of Genes Regulated By Rrp6p, Mary Megan Pelkowski, Kevin Callahan

The Review: A Journal of Undergraduate Student Research

Gene expression is a multi-faceted phenomenon, governed not only by the sequence of nucleotides, but also by the extent to which a particular gene gets transcribed, how the transcript is processed, and whether or not the transcript ever makes it out of the nucleus. Rrp6p is a 5’-3’ exonuclease that can function independently and as part of the nuclear exosome in Saccharomyces cerevisiae (Portin, 2014). It degrades various types of aberrant RNA species including small nuclear RNAs, small nucleolar RNAs, telomerase RNA, unspliced RNAs, and RNAs that have not been properly packaged for export (Butler & Mitchell, 2010). This exosome …

Sickle Cell Anemia: Current Treatments And Potential Advancements, Vanessa Martinez

Selected Honors Theses

Sickle cell anemia is a disease that affects red blood cells, specifically the hemoglobin protein. An amino acid mutation in the gene that encodes β-globin leads to malformation of the β subunit of hemoglobin.3, Valine becomes glutamic acid in the mutated β-globin gene.9 The mutation malformation leads to the red blood cells becoming sickle shaped, or crescent shaped.3,4 The sickle shape of red blood cells in individuals with sickle cell disease leads to vaso-occlusive crisis.9 Vaso-occlusive crisis includes complications such as blood clotting, chronic pain, organ failure, organ death, and possibly early mortality.9 The Centers for Disease Control and Prevention …

Crispr/Cas9 Genome Editing System And Its Use In Infectious Disease, Dustin Daws

Thinking Matters Symposium Archive

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is an exciting new genetic engineering technology that was found in the chromosomes of certain bacteria and archaea. Bacteriophage are viruses that insert their DNA and hijack the host cell’s machinery to make new phage that can go on to infect new cells. Some microorganisms use CRISPR as a defense mechanism to disrupt the bacteriophage DNA after it is inserted into the cell. CRISPR/Cas9 uses genome editing as a means to alter very specific sections of a foreign genome. CRISPR works with a RNA-guided DNA endonuclease called Cas9. This enzyme can locate and …

Rolling Circle Mutagenesis Of Gst-Mcherry To Understand Mutation, Gene Expression, And Regulation, Jessica Cole, Amanda Ferguson, Veronica A. Segarra, Susan Walsh

Faculty Publications

Undergraduates are often familiar with textbook examples of human mutations that affect coding regions and the subsequent disorders, but they may struggle with understanding the implications of mutations in the regulatory regions of genes. We have designed a laboratory sequence that will allow students to explore the effect random mutagenesis can have on protein function, expression, and ultimately phenotype. Students design and perform a safe and time-efficient random mutagenesis experiment using error-prone rolling circular amplification of a plasmid expressing the inducible fusion protein glutathione S-transferase (GST)-mCherry. Mutagenized and wild-type control plasmid DNA, respectively, are then purified and transformed into bacteria …

Overcoming Hurdles To Development Of A Vaccine Against Pneumocystis Jirovecii, Beth A. Garvy

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Development of Pneumocystis pneumonia (PCP) is a common problem among immunosuppressed individuals. There are windows of opportunity in which vaccination would be beneficial, but to date, no vaccines have made it to clinical trials. Significant hurdles to vaccine development include host range specificity, making it difficult to translate from animal models to humans. Discovery of cross-reactive epitopes is critical to moving vaccine candidates from preclinical animal studies to clinical trials.

Phylogenetic Studies Of The Madagascan Freshwater Crabs (Potamoidea, Potamonautidae, Deckeniinae), Rainee Stevens

All NMU Master's Theses

Relationships within the Madagascan freshwater crab fauna were examined based on unstudied specimens from all parts of Madagascar. This study allowed the examination of the validity of existing genera and species, and identified potential new taxa. The new specimens provided phylogenetic and evolutionary data, as well as new insights into the distribution patterns of the Malagasy freshwater crab fauna. In addition, the large number of new localities from previously unsurveyed areas of Madagascar allowed the construction of updated distribution maps.

The present analysis included 62 unidentified specimens plus 13 identified species of Malagasy freshwater crab taxa that had already been …

Topological And Thermodynamic Factors That Influence The Evolution Of Small Networks Of Catalytic Rna Species, Jessica Anne Mellor Yeates, Philippe Nghe, Niles Lehman

Chemistry Faculty Publications and Presentations

An RNA-directed recombination reaction can result in a network of interacting RNA species. It is now becoming increasingly apparent that such networks could have been an important feature of the RNA world during the nascent evolution of life on the Earth. However, the means by which such small RNA networks assimilate other available genotypes in the environment to grow and evolve into the more complex networks that are thought to have existed in the prebiotic milieu are not known. Here, we used the ability of fragments of the Azoarcus group I intron ribozyme to covalently self-assemble via genotype-selfish and genotype-cooperative …

P02. Production Of A Protein Subunit Vaccine For Mannheimia Haemolytica In Lettuce Chloroplasts, Coby K. Martin

Western Research Forum

The cattle industry worldwide is ravaged by bovine respiratory disease (BRD), a bacterial disease caused by Mannheimia haemolytica. We have designed a chimeric protein subunit vaccine against M. haemolytica based on recent evidence demonstrating the protective potential of antigens against a virulence factor, leukotoxin, in addition to a surface lipoprotein. Plant-based production of this protein vaccine provides a safe and inexpensive alternative to traditional production methods. Plant-base production also supports the use of an edible vaccine that will deliver antigens to pharyngeal tissues to provide local immunization against M. haemolytica to prior to its progression into the lungs. Chloroplasts …

Patterns Of Morphological And Molecular Evolution In The Antillean Tree Bat, Ardops Nichollsi (Chiroptera: Phyllostomidae), Roxanne J. Larsen, Peter A. Larsen, Caleb D. Phillips, Hugh H. Genoways, Gary G. Kwiecinski, Scott C. Pedersen, Carleton J. Phillips, Robert J. Baker

University of Nebraska State Museum: Mammalogy Papers

Species endemic to oceanic islands offer unique insights into the mechanisms underlying evolution and have served as model systems for decades. Often these species show phenotypic variation that is correlated with the ecosystems in which they occur and such correlations may be a product of genetic drift, natural selection, and/or environmental factors. We explore the morphologic and genetic variation within Ardops nichollsi, a species of phyllostomid bat endemic to the Lesser Antillean islands. Ardops nichollsi is an ideal taxon to investigate the tempo of evolution in Chiroptera, as it: is a recently derived genus in the family Phyllostomidae; contains …

Draft Genome Sequences Of Three Closely Related Isolates Of The Purple Nonsulfur Bacterium Rhodovulum Sulfidophilum, Michael S. Guzman, Beau Mcginley, Natalia Santiago-Merced, Dinesh Gupta, Arpita Bose

Biology Faculty Research

We report here the draft genome sequences of three isolates of Rhodovulum sulfidophilum from a single population that will serve as a model system for understanding genomic traits that underlie metabolic variation within closely related marine purple nonsulfur bacteria in natural microbial communities.