Genetics and Genomics Commons^™

Investigation Of The Individual Soybean Root Nodule Microbiome, Parris Mica Mayhood

Graduate Theses/Dissertations

Soybean is a valuable crop that establishes a symbiotic association with nitrogen-fixing microorganisms. Previous research suggests members of the genus Bradyrhizobium are the major nitrogen-fixing inhabitants of soybean root nodules. However, results from recent studies identified the presence of other bacteria, so called non-rhizobial endophytes (NREs), inside root nodules. All previous research has pooled nodules of different plants for bacterial community analysis, but this study investigated the presence and relative abundance of rhizobia and NREs within multiple root nodules of a single plant. Three sites were selected on Kendrick Farm in Springfield, MO, where three plants and rhizosphere soil were …

Effect Of Low Dose Irradiation On Expression Of Genes Involved In Ethylene Biosynthesis Of 'Granny Smith' Apples, Brian Nyakundi

Food Science (MS) Theses

Superficial scald is a physiological disorder characterized by skin browning that appears during or after storage of ‘Granny Smith’ apples. The mechanism for scald development is not fully understood, but it is hypothesized that scald formation is related to increased ethylene production. In apples, low dose irradiation induced ethylene reduction appears to be directly related to prevention of superficial scald. However, the molecular basis of such an irradiation induced effect is not known. In this study, effort was made to better understand the molecular basis of the decrease in ethylene production in apple fruit after treatment with x-ray irradiation. ‘Granny …

Smrt Sequencing Of Paramecium Bursaria Chlorella Virus-1 Reveals Diverse Methylation Stability In Adenines Targeted By Restriction Modification Systems, Samantha R. Coy, Eric R. Gann, Spiridon E. Papoulis, Michael E. Holder, Nadim J. Ajami, Joseph F. Petrosino, Erik R. Zinser, James L. Van Etten, Steven W. Wilhelm

James Van Etten Publications

Chloroviruses (family Phycodnaviridae) infect eukaryotic, freshwater, unicellular green algae. A unique feature of these viruses is an abundance of DNA methyltransferases, with isolates dedicating up to 4.5% of their protein coding potential to these genes. This diversity highlights just one of the long-standing values of the chlorovirus model system; where group-wide epigenomic characterization might begin to elucidate the function(s) of DNA methylation in large dsDNA viruses. We characterized DNA modifications in the prototype chlorovirus, PBCV-1, using single-molecule real time (SMRT) sequencing (aka PacBio). Results were compared to total available sites predicted in silico based on DNA sequence alone. SMRT-software detected …

Identifying Phenotypic Effects In Sporisorium Reilianum And Ustilago Maydis Lacking The Alternative Oxidase Gene., Caroline D. Culver

College of Arts & Sciences Senior Theses

Ustilago maydis and Sporisorium reilianum are both dimorphic fungi that cause infection in the host plant, corn (Zea mays). In order for infection to be successful, compatible haploid mating types must form a dikaryon that later leads to the development and dispersal of teliospores. In order to sustain growth, energy is produced via the electron transport chain within the mitochondrion. Host plants often produce harsh reactive oxygen species, resulting in the need to use an alternative respiration pathway such as employing an alternative oxidase. Therefore, this study attempted to investigate the role of the aox gene in pathogenicity …

Comparative Genomics Of Human Mesenchymal Stem Cells And Human Mesenchymal Stem Cells Derived Vascular Smooth Muscle Cells, Samia Ismail

Biomedical Engineering Undergraduate Honors Theses

As of 2017, vascular diseases contributed to 23.1% of all deaths in America. To address the need for more effective and sustainable treatment options for these ailments, stem cell differentiation and implantation has emerged as a viable alternative to standard bypass and graft insertions. A completely autologous treatment can be achieved by extracting adult stem cells, differentiating them into vascular smooth muscle cells (VSMCs), and then reimplanting these cells at the affected tissue site. This study aims to investigate the efficiency of the VSMC differentiation from human mesenchymal stem cells (hMSCs) by comparing 4 cell lines of untreated hMSCs with …

The Significance Of Mfd And Oxidative Damage In Stationary-Phase Mutagenesis In Bacillus Subtilis, Holly Anne Martin

UNLV Theses, Dissertations, Professional Papers, and Capstones

The process of stationary-phase mutagenesis, also known as adaptative or stress-induced mutagenesis, is a phenomenon where bacterial cells accumulate mutations in non-replicative conditions. This process has mainly been studied in Escherichia coli and Bacillus subtilis; however, the underlying mechanisms found in each of these systems differ. Here, I use B.msubtilis to study previously understudied aspects of stationary-phase mutagenesis. In this dissertation, I describe work that has led to three major discoveries which are described below.

First in B. subtilis, Mfd is important for stationary-phase mutagenesis and its mutagenic function at regions of the genome that are transcriptionally upregulated has been …

Editing Fusarium Graminearum Using Crispr/Cas9, Grace Marie Sack

Honors Program Theses

Fusarium graminearum is a pathogenic fungus that causes scab or Fusarium head blight disease in barley and wheat. Alternative approaches to protect barley from this devastating disease are badly needed. In recent years, an RNA-guided genome-editing system known as clustered regularly interspaced short palindromic repeat and CRISPR-associated protein 9 (CRISPR/Cas9), has become a popular method for genome modification (Jinek et al., 2012). We applied CRISPR/Cas9 technology in an effort to make F. graminearum less infectious. We used CRISPR/Cas9 plasmids constructed by a former UNI student (Akers, 2019) to mutate three genes in F. graminearum. One of the genes ( …

Identification Of Genome-Linked Bacillus Bacteriophage Proteins, Katherine Dyer

Honors Program Theses

Small viruses that can infect bacteria, called bacteriophages, can be found in the soil. These viruses are being examined as a potential treatment against bacterial infections when antibiotics are not available or non-effective. Before viruses can be used as a medical treatment, they must be studied extensively. Previous attempts to sequence the terminal end of the genome of a bacteriophage QCM-11 (Quartz-Crystal-Microbalance-11: Q11) resulted in error. It is thought that a covalently bound protein may be attached to Q11’s DNA, blocking sequencing of the genome from the 5’ end. Proteins bound to the DNA of phages is not unheard of, …

Construction Of A Target-Specific Crispr-Cas9 Vector For The Purpose Of Introducing Mutations Into The Fum1 Gene Of Fusarium Verticillioides, Jennifer Petsche

Honors Program Theses

In molecular biology, CRISPR-Cas9 is a useful gene editing tool that takes advantage of a powerful cleaving enzyme and DNA sequences found in prokaryotes. In this study, a target-specific CRISPR-Cas9 vector containing an RNA-guided Cas9 nuclease specific for the FUM1 gene was constructed for the purpose of introducing mutations into the FUM1 gene of Fusarium verticillioides. Plasmids pFC332 and pFC334 were isolated. A guide RNA sequence was designed and amplified using Polymerase Chain Reaction (PCR) with pFC334 as the template. Sticky ends were created through restriction enzyme digestion of pFC332. These pieces were combined using Uracil-Specific Excision Reagent (USER) …

The Effects Of Intercalating Molecules On The Polymer Properties Of Dna, Joseph Tibbs

Honors Program Theses

This project used Atomic Force Microscopy (AFM) imaging to examine the effects of different intercalation conditions upon the elastic properties of intrinsically straight DNA molecules. Three intercalating molecules (chloroquine, ethidium bromide, and acridine) were used to demonstrate the dependence of persistence length and mean polymer extension on the intercalator type. It was found that all three intercalators significantly increased persistence length, and that this effect was most pronounced for ethidium bromide. In addition, an analysis of the normal bending modes of the static molecules corroborated these results. The only intercalator which resulted in significant extension of the DNA polymers was …

Assessing The Influence Of Riparian Invasion By The Shrub Lonicera Maackii On Terrestrial Subsidies To Headwater Streams, Erin C. Rowekamp, Julia I. Chapman, Ryan Mcewan

Biology Faculty Publications

Invasion of Amur honeysuckle (Lonicera maackii) into riparian areas of headwater streams creates strong potential for alterations of terrestrial subsidies that may drive bottom-up effects on aquatic biota. In this study, we analyzed effects of L. maackii on terrestrial subsidies in stream sites that represented a gradient of invasion intensity in temperate deciduous forests of southwestern Ohio (USA). Leaf litter biomass, terrestrial and aquatic fine woody debris (0.5–9.9 cm diameter) volume and count, and terrestrial and aquatic coarse woody debris (>9.9 cm diameter) volume were analyzed. Based on previous research, we hypothesized that sites with a higher …

Meta-Analysis Of Biological Research Literature, Evan Suggs

Honors Theses

Comparative studies have been powerful tools in generating a broad understanding about the evolution of animal social systems but they currently rely on the slow, manual process of reading thousands of abstracts and papers from research databases. A web application was created for researchers conducting a comparative survey, in order to speed up their research. This web application automates the retrieval of research papers and their selection process. Using previously obtained data sets on the orders Artiodacytla and Lagomorph, a machine learning application was created to classify the papers. These techniques and tools should greatly increase the speed at which …

Discrimination Of Monozygotic Twins Using Dna Methylation Levels Of One Cpg Site At Chromosome 3, Dino O. Robinson

Student Theses

Conventional STR typing, commonly used in forensics for human identification, poses a problem in criminal cases and paternity disputes involving monozygotic (MZ) twins because they share identical DNA sequences. To date, no routine method is available in forensics to differentiate between individuals of MZ pairs. Recently, epigenetic methods measuring differential DNA methylation patterns have been applied to MZ twin differentiation. In this study, we investigated the potential to identify MZ twins using a previously identified DNA methylation site in chromosome 3, cg18562578, in a sample of 129 MZ and 37 dizygotic (DZ) twin pairs. We used bisulfite converted saliva DNA …

Characterization Of A Mycoplasma Pneumoniae Cards Toxin Mutant, Nikaash Pasnoori

Honors Scholar Theses

Mycoplasma pneumoniae is a high-burden pathogen which causes mild to significant infections of the respiratory system. According to the CDC, an estimated two million cases occur yearly in the United States alone, demonstrating the widespread effect of the pathogen. In addition to being the cause of respiratory infections, M. pneumoniae has also been implicated in exacerbating pre-existing asthma conditions. These morbidities make finding a vaccine candidate a vital part of easing the healthcare burden caused by the pathogen. The current mechanism of infection is unknown, but recent evidence points to the Community Acquired Respiratory Distress Syndrome (CARDS) toxin as being …

Identification And Analysis Of Feather Degrading Bacteria: A Search For Keratinase Genes, Nehal Navali

Honors Scholar Theses

Over two million tons of feather waste is generated annually by the poultry industry, the majority of which goes into landfills due to the difficulty of degrading its major component keratin. Although a portion of feather waste is eliminated via incineration or chemical treatment, the use of Feather Degrading Bacteria (FDB) has been proposed as a cheap and eco-friendly alternative. FDBs have been consistently isolated from the feather microbiome of birds and contain genes coding for the specialized protein keratinase which is able to degrade feathers. By doing so, feather waste, which is rich in nutrients, can be repurposed as …

Antibiotic Resistant Bacteria, Antibiotic Resistance Genes And Potential Drivers In The Aquatic Environments, Shuo Shen

Dissertations

As antibiotic resistance genes in aquatic environment have been increasing across the world, affecting water quality and public health, many studies documented concentrations of antibiotic resistance genes and some studies discussed their potential drivers. However, systematic and quantitative reviews that link antibiotic resistance genes (ARGs) to anthropogenic and environmental factors are limited. Nevertheless, this information will be important for developing regulation policy on controlling antibiotic use and therefore reducing potential risks to antibiotic resistance. I conducted meta-analysis of ARGs concentration at a global scale using Bayesian inference to explore climatic and socio-economic factors as drivers. I found local-scale climatic variables …

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 …

Genetic Variation Of Clonal Ananas Comosus Var. Md2 Plants After Mutagenic Treatment At Different Post-Recovery Periods, Mohd Raffi Anis Norsyahira

Student Works (2020-2029)

Studies on genetic variation in plants had become extensively employed in research for plant improvement and conservation. Due to their sessile nature, plants are exposed to various environmental stressors such as exposure to enhanced levels of ultraviolet (UV), ionizing and non-ionizing radiations. Exposure to these harmful radiations will result in various damages, ranging from DNA and chromosomal aberrations to phenotypic abnormalities. However, plants have evolved efficient DNA repair mechanisms to detect and repair any DNA damages caused by the exposure to these harmful stressors, therefore ensuring their survival. In this study, the effect of gamma radiation (as a source of …

Multiplexed Crispr Libraries For Cancer Functional Genomics, Jintan Liu

Dissertations and Theses (Open Access)

High-throughput forward genetic screenings are invaluable tools to systematically explore genetic interactions and to link gene disruption with disease contexts. The adaptation of CRISPR/Cas9 has improved the sensitivity and specificity of functional screenings. Despite this advance, there remains a long-standing need to improve functional screenings with smaller and more versatile pooled libraries. Capitalizing on the inherent multiplexing capability of a class 2 CRISPR enzyme AsCpf1, we developed a multiplexed, high throughput screening strategy that has avoided the usual trade-off between library size and library penetration, allowing library minimization without sacrificing gene targeting efficiency. We optimized the AsCpf1 protein for functional …

Factors That Impact Uptake Of Carrier Screening By Male Reproductive Partners Of Female Prenatal Patients, Wendi Betting

Dissertations and Theses (Open Access)

Carrier screening is a genomic technology that is used to identify individuals who are carriers of autosomal recessive conditions. Despite published recommendations, the majority of male partners do not complete carrier screening after their female partner is identified to be a carrier. Previous studieshave examined reasons why women elect or decline carrier screening, but there have been few published studies that examine factors that influence a male partner’s decision to elect or decline carrier screening, particularly when the female has been identified as a carrier. The aim of the study was to determine the factors that influence the uptake of …

Heterogeneous Nuclear Ribonucleoprotein K (Hnrnp K) Overexpression And Its Interaction With Runx1 Rna In Acute Myeloid Leukemia, Marisa Aitken

Dissertations and Theses (Open Access)

Acute myeloid leukemia (AML) is an often devastating hematologic malignancy with 5-year overall survival lingering near 20%. Acquiring a deeper understanding of molecular underpinnings of leukemogenesis will provide a basis for developing more effective therapeutic strategies for patients with AML.

Here, we identified overexpression of hnRNP K as a recurrent abnormality in a subset (~20%) of AML patients. High levels of this RNA-binding protein associated with inferior clinical outcomes in de novo AML. Thus, to evaluate its putative oncogenic capacity in myeloid disease, we overexpressed hnRNP K in murine hematopoietic stem and progenitor cells isolated from fetal liver cells (FLCs). …

Loss Of Caspase-8 Function In Combination With Smac Mimetic Treatment Sensitizes Head And Neck Squamous Carcinoma To Radiation Through Induction Of Necroptosis., Burak Uzunparmak

Dissertations and Theses (Open Access)

Caspase-8 (CASP8) is one of the most frequently mutated genes in Head and Neck Squamous Carcinomas (HNSCC), and mutations of CASP8 are associated with poor overall survival. The distribution of these mutations in HNSCC suggests that they are likely to be inactivating. Inhibition of CASP8 has been reported to sensitize cancer cells to necroptosis, a unique cell death mechanism. Here, we evaluated how CASP8 regulates necroptosis in HSNCC using cell line models and syngeneic mouse xenografts. In vitro, knockdown of CASP8 rendered HNSCCs susceptible to necroptosis induced by a second mitochondria-derived activator of caspase (SMAC) mimetic, Birinapant, when combined …

Liver Transcriptomic Analysis After Short- And Long-Term Feeding Of Soy Protein Isolate And Its Ability To Reduce Liver Steatosis In Obese Zucker Rats, Melisa Kozaczek

Graduate Theses and Dissertations

According to the Centers for Disease Control and Prevention (CDC) the prevalence of obesity in adults in the United States during 2017-2018 was a 42.4%, a high number considering all the risks factors associated with this disorder, such as cardiovascular disease, insulin resistance, diabetes type 2, and fatty liver disease, among others. Fatty liver disease is the accumulation of lipids in the liver that can account for more than 5 to 10% of the liver’s weight. There are two types of fatty liver disease, alcoholic fatty liver disease (AFLD), and non-alcoholic fatty liver disease (NAFLD). AFLD is the detrimental accumulation …

Characterization Of The Broad-Spectrum Inhibitory Capability Of Alcaligenes Faecalis And A. Viscolactis Against Potential Pathogenic Microorganisms, Andrew Fuqua

Undergraduate Honors Theses

The recent rise of multidrug resistant microorganisms has grown from an isolated concern to a massive public health crisis. It has become imperative that scientists look for new ways to combat this issue. Due to the selective pressures of competition, bacteria and other microbes possess a host of defenses and weapons designed to exploit vulnerabilities in other microorganisms. Consequently, the study of these systems and microbial interactions has much to reveal in the search for novel antimicrobial treatments. Previous research from our laboratory has discovered that both Alcaligenes faecalis and Alcaligenes viscolactis, two rarely studied and generally non-virulent bacteria, …

Subcellular Localization Of Tobacco Sabp2 Under Normal And Stress Conditions, Sanjeev Das

Undergraduate Honors Theses

Subcellular Localization of Tobacco SABP2 under Normal and Stress Conditions

Salicylic acid (SA), a phytohormone, plays an important role in plant physiology. SA mediated innate immune pathway is an important pathway for plant immunity against pathogens. Plants resisting pathogen infection synthesize higher levels of Methyl Salicylate (MeSA), which is then converted to SA by the esterase activity of Salicylic Acid Binding Protein 2 (SABP2). The high level of the converted SA leads to enhanced pathogen resistance. The study of subcellular localization of a protein is critical in explaining its potential biochemical functions. SABP2 tagged with eGFP was expressed transiently in …

A Review Of The Characteristics Of Super-Utilizers And Evidence-Based Approaches To Reduce Healthcare Utilization, Suswara Mandala Rayabandla

Capstone Experience: Master of Public Health

A significant fraction of health care resources in the United States of America is utilized by a comparatively small number of people. An examination of the attributes and patterns and associated evidence-based interventions for such high-utilizing patients might aid clinicians to improve interventions to address the distinctive needs of these patients, decrease their risks for numerous hospitalizations, and contribute to reducing the costs. This study aims at exploring the existing literature on the characteristics of super-utilizers and interventions to reduce avoidable use of health care among this population. The method used for this research is a comprehensive literature review. Search …

Differences In Learning And Gene Expression In Brains Of Male And Female Bicyclus Anynana, Gabrielle Agcaoili

Biological Sciences Undergraduate Honors Theses

One way to understand the variation in the behavior of animals is by looking at the genes involved. We were particularly interested in behavioral differences between the sexes. How might these differences be manifested in the brain? This study worked to answer this question by using male and female butterflies of the species Bicyclus anynana, examining what is going on in the brains of males and females as they learned from a social exposure. We focused in on how sex plays a role in behavior and learning; it has been seen that males and females respond to the same social …

Understanding Cytochrome C Maturation In Anaerobic Archaea, Blake Wojciechowski

Biological Sciences Undergraduate Honors Theses

Methanoperedens nitroreducens (MPEBLZ), an archaeal methanotroph and close relative of Methanosarcina acetivorans, contain numerous cytochrome c proteins. However, difficulty in using these organisms as a model for cytochrome cresearch has created a pressure to express cytochrome c proteins in an organism that is much easier to work with. A punitive cytochrome c protein (MPEBLZ_04274) from M. nitroreducens was attempted to be cloned into a model methanogen M. acetivorans as well as Escherichia coli. Cytochrome c proteins are important for many metabolic processes within anaerobic archaea. In order for a mature cytochrome c to be formed heme must …

Development Of Arkansas Net Energy Equation, Katie Hilton

Graduate Theses and Dissertations

The modern broiler is growing at a rapid rate generating tremendous amounts of heat. A sensitive Net Energy (NE) system is needed to measure body heat production (HP) generated primarily by daily maintenance and synthesis and degradation of myofibrillar and sarcoplasmic protein. The first two chapters present evaluation of the classic way to calculate NE versus a new methodology; the Arkansas NE (Ark NE) system, with birds from two genetic lines fed diets with different AA content or different ME content in two different environmental temperatures.

Utilizing together the Net Energy maintenance (NEm), determined from indirect calorimetry, and Net Energy …

Evolutionary Inference From Admixed Genomes: Implications Of Hybridization For Biodiversity Dynamics And Conservation, Tyler Chafin

Graduate Theses and Dissertations

Hybridization as a macroevolutionary mechanism has been historically underappreciated among vertebrate biologists. Yet, the advent and subsequent proliferation of next-generation sequencing methods has increasingly shown hybridization to be a pervasive agent influencing evolution in many branches of the Tree of Life (to include ancestral hominids). Despite this, the dynamics of hybridization with regards to speciation and extinction remain poorly understood. To this end, I here examine the role of hybridization in the context of historical divergence and contemporary decline of several threatened and endangered North American taxa, with the goal to illuminate implications of hybridization for promoting—or impeding—population persistence in …