Genetics and Genomics Commons^™

Evaluating Threats To The Water Quality Of Little Buffalo Creek And Sarver Run Through Water Quality Analysis And Backpack Electrofishing Surveys, Riley Williams

Electronic Theses and Dissertations

Currently, 64.8% and 52.2% of Little Buffalo Creek and Sarver Run stream kilometers are impaired, respectively. Impairment sources to Little Buffalo Creek include agriculture, riparian deforestation, on-site wastewater, and urban runoff. Impairment sources to Sarver Run are unknown. Our goal was to understand sources of impairment through water quality analysis and fish surveys. Water samples were collected monthly and analyzed for cation and anion concentrations. Mass ratio analysis comparing Br/SO₄ and Mg/Li, Ca/Mg and Ca/Sr, Mg/Na and SO₄/Cl, and SO_4/Cl and Mg/Li suggests that mine drainage and conventional oil and gas may be impacting water …

A Review Of Current Methods In Avian Dietary Analysis And Their Integrated Application To Characterize The Trophic Niche Of Louisiana Waterthrush (Parkesia Motacilla)., Brandon Hoenig

Electronic Theses and Dissertations

Characterizing a species’ dietary composition presents an avenue to understand many facets of its ecological niche and can provide essential information for the species’ long-term conservation. To date, the vast majority of diet studies have relied on direct identification of prey during foraging observations or from diet samples to characterize the dietary habits of birds. However, advancements in laboratory-based approaches have revolutionized the field of trophic ecology by allowing researchers to indirectly infer dietary habits with higher resolution across greater time scales. Here, I apply two of these laboratory-based techniques, namely DNA metabarcoding and stable isotope analysis, to characterize the …

Analysis Of Water Quality And Fish Communities Of Crooked Creek, Tributary To The Allegheny River In Southwestern Pennsylvania, Katherine Stupar

Electronic Theses and Dissertations

Crooked Creek is a tributary of the Allegheny River with a drainage area of 292 square miles. It has been historically affected by mine drainage pollution. Extensive cleanup and monitoring efforts by the Commonwealth of Pennsylvania and the U.S. Army Corps of Engineers (USACE) have taken place over the last four decades. Unconventional oil extraction, or fracking, presents an emerging threat to the water quality of this stream and the health of its fish communities. Here we perform an analysis of the Crooked Creek watershed utilizing backpack electrofishing, eDNA metabarcoding, and various methods of water quality analysis. Seven sites on …

Genomic Tools And Models For Investigating The Role Of Germline Diversity In Mouse Antibody Repertoire Development., Justin T. Kos

Electronic Theses and Dissertations

Given the diversity and complexity within immunoglobulin (IG) loci, effective mouse models first require characterization of intra-strain differences and construction of high-quality reference assemblies for IG loci in several representative strains. To understand light chain germline diversity across biomedically significant mouse strains, we profiled the expressed IGK and IGL repertoires of 18 commonly used laboratory mouse strains using AIRR-seq. Across strains, we observed germline IGKV sequences shared by three different IGK haplotypes and a more conserved IGLV germline repertoire among common laboratory strains. Pacific Biosciences (PacBio) Single-Molecule Real-Time (SMRT) sequencing was used to sequence and assemble bacterial artificial chromosomes (BAC) …

Functional Role Of Ppal And Potential For Moss In Industrial Applications., Susana Perez Martinez

Electronic Theses and Dissertations

This dissertation is an examination and characterization of the functional roles of PPAL. PROTEIN PRENYLTRANSFERASE ALPHA SUBUNIT-LIKE (PPAL) is a recently discovered gene. PPAL homologs are present in all plants and many animals, where its function is largely unknown. It is possible that PPAL could participate in prenylation processes since it shares similarity to the α subunits of known prenylation enzymes. Prenylation is a post-translational modification of proteins that involves the addition of a lipid moiety to proteins to facilitate membrane targeting and association and promote protein-protein interactions. Prenylation has important roles in plant growth and development, including …

Expression And Functional Characterization Of Avocado Dgat1 And Pdat1 In Arabidopsis And Camelina, Josphat Kiunga

Electronic Theses and Dissertations

The study is aimed to determine the role of avocado DGAT1 and PDAT1 in seed oil synthesis. Triacylglycerol (TAG) has a nutritional and industrial value and is essential for plant growth. DGAT1 and PDAT1 catalyze the final step of TAG Assembly. We hypothesized that both PaPDAT1 and PaDGAT1, although predominantly expressed in non-seed tissues, could contribute to oil accumulation in seeds. Agrobacterium transformants with PaPDAT1 and PaDGAT1 cloned in pCAMBIA were generated to test this. Subsequently, the Agrobacterium-mediated transformation of Arabidopsis mutant lines and camelina was carried out by floral dipping. The T₁ camelina seeds …

Characterization Of Hemocytes In Amblyomma Americanum And Investigation Of Traf6’S Role In Proliferation Of Hemocytes In Response To Infection, Jacquelyn T. May

Electronic Theses and Dissertations

Amblyomma americanum is a medically important vector in the southeast United States. Cellular and molecular aspects of the immune system of this species were examined. To investigate cellular processes, hemocytes produced in response to gram-negative infection were characterized according to their ultrastructure. Four hemocyte types were identified: prohemocytes, plasmatocytes, type-I granulocytes, and type-II granulocytes. To elucidate molecular processes, we investigated AamTRAF as an immune-related gene by silencing the gene using RNAi then quantifying the hemocytes after inoculation with gram-negative and positive bacteria. It was found that silencing AamTRAF caused a decrease in hemocyte proliferation in response to gram-positive but not …

Understanding The Impact Of Human Germline Single-Nucleotide Variants, Samuel Adjei

Electronic Theses and Dissertations

All genetic variations in the human genome are due to mutations, and therefore determining the impact of the different categories of mutations, particularly nonsynonymous single nucleotide variants (SNVs) which are the most abundant in the population is essential for understanding the genetics of human diseases. Generations of mutation studies have focused on a relatively small number, partly due to technological limitations. However, advances in next-generation sequencing have allowed for empirical assessments of genome-wide mutations. genome sequence of any individual contains extensive protein-altering genetic variation (missense mutations) and of which only a few are unambiguously deleterious. Characterizing the phenotypes of these …

Crispr/Cas9 Whole-Genome Screens Identify Genes Critical For Neutral Lipid Metabolism In Macrophages, Rifat Sultana

Electronic Theses and Dissertations

Lipid-laden macrophages contribute to atherosclerotic plaque formation in atherosclerosisrelated heart diseases. Thus, understanding formation and degradation of lipid would help identify therapeutic targets for the treatment of atherosclerosis. In the first chapter of this dissertation, I have explained the current understanding of cholesterol metabolism in macrophages and how mTOR could play a potential role in this process. Additionally, I have described how CRISPR/Cas9 whole-genome screens present an effective tool to identify genes regulating cholesterol metabolism including positive and negative regulators of lipid droplet formation and lipid droplet degradation. In Chapter 2, I have described the results of the CRISPR/Cas9 whole-genome …

Understanding Snow Leopard (Panthera Uncia) Population Structure, Diet, And Human-Wildlife Dimensions Using Noninvasive Genetic Approaches, Charlotte Hacker

Electronic Theses and Dissertations

Snow leopards (Panthera uncia) are a large cat endemic to the mountains of Central Asia. Despite their status as a flagship species, there are numerous knowledge gaps surrounding their population status, dietary ecology, and relationship with humans. Such knowledge gaps are prohibitive for effective conservation action for the species. Snow leopards are threatened by habitat loss, decreased prey availability, retaliatory killings, and large-scale negative impacts associated with climate change. In order to help bridge these gaps, noninvasive genetic methodologies were deployed in this work across several disciplines. Population genetic metrics were garnered at local, regional, and range-wide scales, …

Investigation Of Mitochondrial Inheritance In The Smut Fungus Sporisorium Reilianum., Hector Eduardo Mendoza

Electronic Theses and Dissertations

An important goal in evolutionary biology is to address the origin of Earth’s immense biodiversity through the evolution of complex sexual reproduction mechanisms in eukaryotes. Inheritance of mitochondria during sexual reproduction has received special attention in recent years, as these organelles cannot be synthesized de novo and must be transmitted from parent to offspring. The importance of these organelles far exceeds its common function as the energy-producing “powerhouse” of the cell, as it has been found to also be involved in fundamental processes like apoptosis, aging and metabolic homeostasis. Thus, appropriate inheritance of mitochondria is essential for growth and development …

Effects Of Respiratory Perturbations On Aging And Healthspan In Daphnia Magna, Millicent Nkiruka Ekwudo

Electronic Theses and Dissertations

Aging is a degenerative process characterized by a decline in physiological functions and cellular activities. Environmental and pharmacological interventions affecting longevity pathways have been extensively studied in model organisms. This study investigated the effect of chronic mild intermittent hypoxia (4 mg O2/L) or mild mitochondrial uncoupling with three doses of 0 (control), 0.1, 1, and 5 μM of 2,4-Dinitrophenol (DNP), on life history and gene expression in four clones of Daphnia magna. Interestingly, clones from intermittent ponds displayed better tolerance to hypoxia and DNP. Although neither treatments extended longevity, hypoxia increased fecundity and body size, and decreased food consumption and …

An Evaluation Of The Bighorn Sheep Population In Badlands National Park, Austin J. Wieseler

Electronic Theses and Dissertations

Within the last century, bighorn sheep (Ovis canadensis) in the badlands ecosystem of western South Dakota have been subjected to complete extirpation, reintroduction, disease die-offs, genetic bottlenecking, and population augmentation. Subsequently, the population in Badlands National Park (BNP) appears to have recovered, but it was unknown to what degree past events had influenced the population. From 2017-2019, we conducted research on 5 subherds within 2 management units in BNP to 1) survey for the presence of respiratory pathogens and estimate the prevalence of other potentially infectious diseases; 2) assess adult and lamb survival and cause-specific mortality; 3) estimate population size …

Comparison Of Software Packages For Detecting Differentially Expressed Genes From Single-Sample Rna-Seq Data, Rong Zhou

Electronic Theses and Dissertations

RNA-sequencing (RNA-seq) has rapidly become the tool in many genome-wide transcriptomic studies. It provides a way to understand the RNA environment of cells in different physiological or pathological states to determine how cells respond to these changes. RNA-seq provides quantitative information about the abundance of different RNA species present in a given sample. If the difference or change observed in the read counts or expression level between two experimental conditions is statistically significant, the gene is declared as differentially expressed. A large number of methods for detecting differentially expressed genes (DEGs) with RNA-seq have been developed, such as the methods …

Population Genetic Structure Of Bobcats (Lynx Rufus) In South Dakota: Using Harvested Samples To Inform Management, Stuart C. Fetherston

Electronic Theses and Dissertations

A primary objective of state wildlife management agencies is to establish sustainable harvest levels for game species. An important component of sustainable management practices is the identification of appropriate management units for monitoring and establishing defensible harvest levels. Across their range, bobcats (Lynx rufus) are an ecologically and economically important species. Despite their importance, little is known about the genetic structure of bobcat populations in South Dakota. We used tissue sampled from n = 1,215 bobcats harvested across the state from 2014–2019 to infer population genetic structure. We used 17 microsatellite loci and a sex identification marker to assign individuals …

Characterization Of Adiposity And Inflammation Genetic Pleiotropy Underlying Cardiovascular Risk Factors In Hispanics., Mohammad Yaser (Anwar)

Electronic Theses and Dissertations

The observed overlap between genetic variants associated with both adiposity and inflammatory markers suggests that changes in both adiposity and inflammation could be partially mediated by common pathways. The pervasive but sparsely characterized “pleiotropic” genetic variants associated with both adiposity and inflammation have been hypothesized to provide insight into the shared biology. This study explored and characterized the genetic pleiotropy underpinning adiposity and inflammation using genetic and phenotypic observations from the Cameron County Hispanic Cohort (CCHC). A total of 3,313 samples and >9 million single nucleotide polymorphisms (SNPs) were examined in this study. Mixed model genome-wide association studies (GWAS) were …

Statistical Approaches Of Gene Set Analysis With Quantitative Trait Loci For High-Throughput Genomic Studies., Samarendra Das

Electronic Theses and Dissertations

Recently, gene set analysis has become the first choice for gaining insights into the underlying complex biology of diseases through high-throughput genomic studies, such as Microarrays, bulk RNA-Sequencing, single cell RNA-Sequencing, etc. It also reduces the complexity of statistical analysis and enhances the explanatory power of the obtained results. Further, the statistical structure and steps common to these approaches have not yet been comprehensively discussed, which limits their utility. Hence, a comprehensive overview of the available gene set analysis approaches used for different high-throughput genomic studies is provided. The analysis of gene sets is usually carried out based on …

Genes Involved In Nitrogen Assimilation In Ustilago Maydis., Sunita Khanal

Electronic Theses and Dissertations

U. maydis can utilize nitrate as a source of nitrogen from its environment, a process requiring a nitrate transporter to bring the nitrate inside the cell. This is followed by a two-step reduction of nitrate to ammonium. At first, nitrate reductase reduces nitrate into nitrite; in the second reaction, nitrite is reduced to ammonium by nitrite reductase. The goal of the study was to identify the nitrate transporter and nitrite reductase genes in U. maydis and characterize them based on their role in mating and virulence. The genes um03848 and um03849 were identified bioinformatically as encoding, respectively, the nitrite reductase …

Comparative Secretomics And Functional Analysis Of Effectors Utilized By The Microbotryum Genus Of Anther-Smut Fungal Pathogens, And Their Role In Host-Specificity., William Christopher Beckerson

Electronic Theses and Dissertations

Understanding how pathogens evolve in response to changes in their host is paramount to combating the spread of emergent strains of disease. This is particularly true for plant pathogens that cause billions of dollars of damages to crops globally, every year. Understanding the molecular interactions between pathogens and their hosts therefore sheds light on the coevolutionary arms race that can result in host-specificity and host-shifts in plant pathogens. This research approaches the question of how fungal pathogens interact with their plant hosts utilizing both unique and shared arsenals of secreted proteins (SPs) during infection, and addresses the question of whether …

A Genomic Analysis Of Bobcat Populations In North America With A Comparison To The Canada Lynx: An Assessment Of Local Adaptation To Unique Ecoregions And Phylogeography, Jennifer C. Broderick

Electronic Theses and Dissertations

Bobcats (Lynx rufus) are an ecologically and genetically diverse species with a large contiguous range throughout North America. The species not only has a wide array of phenotypic variation compared to other mammals, but shows marked adaptability across ecozones with differing ecological influences. It is these various selective pressures in distinctive parts of the continent that have likely led to localized adaptations within the bobcat metapopulations. The species is also marked by its ability to maintain connectivity and populations in anthropogenically developed areas, an advantage it has over other felids, including its close relative the Canada lynx ( …

Efficient Conditional Inactivation Of Rdh10 Reveals Important Role Of Retinoic Acid During Lung Branching Morphogenesis., Nhut Quang Huy Tran

Electronic Theses and Dissertations

Birth defects are complicated pathological processes. Many birth defects can be traced back to disrupted regulation of molecular signaling pathways during embryogenesis. One of the most important gene regulatory factors during embryo development is retinoic acid (RA). However, studying the specific roles of RA in each organ during embryogenesis in vivo is challenging. The Sandell laboratory has a conditional mutant mouse model that can potentially be used for studying the role of RA during embryogenesis by inducing stage-specific RA deficiency. This model allows retinol dehydrogenase 10 (Rdh10), a gene required for RA production, to be inactivated at a …

Human Genetics, Psychotropic Drugs, And Acts Of Violence, Selma Jolanda Eikelenboom-Schieveld

Electronic Theses and Dissertations

From the start of the use of psychoactive prescription medications in the 1950s, physicians reported paradoxical adverse reactions, ranging from newly developing depressions to an increase in existing mood disorders, and extremely violent and bizarre acts of suicide and homicide. In this research, it is hypothesized that the pharmacological properties of the prescribed drugs or the interaction between the drugs and the enzymes that are primarily responsible for their metabolism (cytochrome P450s) could cause these reactions. Given that acts of violence could be medication-induced, the role of the rate of drug metabolism is discussed. Genetic testing of certain CYP450s could …

In Situ Analysis Of Line-1 Promoter Activity Using Lacz Transgenic Mice, Partha Sarathi Saha

Electronic Theses and Dissertations

Apart from an evolutionary role, transposable elements have been implicated in animal development and also in pathophysiology. Non-LTR retrotransposons– LINE-1, Alu and SVA - are responsible for over 120 cases of human genetic diseases as heritable insertions, and are emerging as an important etiological factor for cancer and neurological disorders as somatic mutations. It is estimated that among the total number of 500,000 LINE-1s presents in the human genome, 80-100 LINE-1s remain competent for retrotransposition. Retrotransposition is only possible when LINE-1 is expressed. Because LINE-1 transcription is regulated by its 5’UTR promoter, it is essential to understand the spatiotemporal control …

Functional Analysis Of Squamosa-Promoter Binding Protein Like 13 In Controlling Flowering Time, Plant Architecture, Grain Size, And Grain Number In Wheat, Ajay Gupta

Electronic Theses and Dissertations

Higher and stable yield is always a major objective of wheat genetic improvement programs. The SQUAMOSA-promoter binding protein-like (SPL) genes constitute a small family of plant-specific transcription factors with diverse functions in plant development and growth and have great potential in improving yield and other major agronomic traits. However, the functional characterization of the SPL gene family in wheat is far behind other cereal crops such as rice. Using phylogenetic analysis, we identified 56 wheat orthologues of rice SPL genes belonging to 19 homoeologous groups. Among these 19 orthologous TaSPL genes, nine harbor the micro RNA 156 recognition element (MRE) …

Examining Biodiversity Metrics And The Utility Of Dna Barcoding In The Northern Great Plains, Sarah Herzog

Electronic Theses and Dissertations

Due to heavy threats to remaining global floral and faunal diversity, it is imperative we can identify species and quantify ecosystem health to find best practices for land management and conservation. The tallgrass prairies of the Northern Great Plains are one example of a heavily imperiled ecosystem. The tallgrass prairies have been reduced to less one percent of their historical extent and are facing continued loss. Genetic approaches and evolutionary theory offer insights for identifying species and assessing how biodiversity metrics may correlate with ecosystem processes. My two projects aim to address two facets imperative to conservation in the tallgrass …

Analysis Of Two Maize Mutations That Arose After Exposure To Radiation From A Plutonium-Beryllium Source And Developing Molecular Markers For Antimorph Aei1-5180, An Alternative To Aei To Increase Amylose Content In Maize Endosperm, Prameela Awale

Electronic Theses and Dissertations

This thesis consists of two independent projects dealing with two different aspects of maize. The first project is about the maize reproduction, where we try to uncover the candidate genes of two mutations that affect pollen viability and successful fertilization. The second project is about maize endosperm starch, where we developed molecular markers to characterize a mutant allele that knocks a major starch branching enzyme in maize.

Analysis of two maize mutations that arose after exposure to radiation from a plutonium-beryllium source
Maize is a monoecious and diclinous plant. Sexual reproduction takes place by the fusion of haploid male and …

Crispr/Cas9 Whole Genome Screens Reveal Novel Regulators Of Endocytic Processes In Macrophages, Jared Wollman

Electronic Theses and Dissertations

Macrophages maintain tissue homeostasis by identifying and eliminating threats within their tissue microenvironment. Pattern recognition receptors allow macrophages to recognize and internalize specific ligands while macropinocytosis allows the internalization of all extracellular solutes from their environment. Without pattern recognition receptors, pathogens may grow unchecked if they cannot be detected, and without macropinocytosis, macrophages struggle to detect and move around their environment. This thesis presents the results of CRISPR/Cas9 whole-genome screens used to identify the regulators of both endocytosis (Chapter 2) and macropinocytosis (Chapter 3). In Chapter 2, we report genes regulating dextran uptake in primary murine macrophages and reveal Mrc1-mediated …

Genetic And Biochemical Analysis Of A Conserved, Multi-Gene System Regulation Spore-Associated Proteins In Streptomyces Coelicolor, Joseph Sallmen

Electronic Theses and Dissertations

Streptomyces coelicolor is a Gram positive, filamentous soil dwelling bacterium that exhibits a complex life cycle including the transition from a vegetative mycelium to a sporulating, aerial mycelium. Early genetic studies identified two classes of genes that resulted in developmental blocks, bald and white. The bald (bld) phenotype occurs when colonies cannot produce aerial hyphae. White (whi) colonies exhibit incomplete sporulation and/or loss of production of the concurrently produced grey pigment. While some of the original mutants have been explored, the developmental functions of many of the identified genes are not well understood. Of particular interest …

Genetic Analysis Of A Novel Ftsk Homolog, Hfka, In Streptomyces Coelicolor Development-Associated Chromosome Segregation, Sumedha Sethi

Electronic Theses and Dissertations

A quintessential phenomenon occurring during prokaryotic development is accurate segregation of the replicated genomes into the daughter cells. Key energy-dependent processes like chromosome condensation and subcellular partitioning of the genomes are driven by conserved proteins like SMC, ParB, FtsK. During its complex developmental cycle, Streptomyces coelicolor segregates its genomes into chains of unicellular spores when its multigenomic syncytial aerial hyphae undergo division.

A novel ftsK-like gene, hfkA (Homolog of FtsK protein A), was examined for function and localization during development-associated chromosome segregation. Individual deletions did not affect segregation, but a ΔhfkA ΔftsK mutant exhibited 8% anucleate …

A Possible Luxr Solo Type Regulator Of An Antibiotic-Like Compound From The Soil Bacterium Rhodococcus, Katelyn Sellick

Electronic Theses and Dissertations

Rhodococcus, a species of bacteria commonly found in the soil, is an under-explored producer of small bioactive compounds including siderophores, pigments and antibiotics. MTM3W5.2 is a strain of Rhodococcus that was previously discovered to produce an antibiotic-like compound that has inhibitory effects on other Rhodococcus strains, including the veterinary pathogen, R. equi. The biosynthetic gene cluster responsible for production of the antibiotic has been identified, and a small gene, BTZ20_3964 at the start of the operon is believed to be a luxR solo regulator of the gene cluster. The goal of this project was to determine this gene’s …