Life Sciences Commons^™

Organelle Genome Evolution Within The Green Algal Genus Dunaliella, Michael Del Vasto

Electronic Thesis and Dissertation Repository

A large amount of diversity in genomic size and content exists within the mitochondrial and plastid genomes of green algae. However, there are still many un-sequenced green algal organelle genomes. In Smith et al. 2010, the green algal species Dunaliella salina, isolated from Western Australia (strain CCAP 19/18), had its organelle genomes fully sequenced. The genomes of this organism were found to contain large amounts of noncoding DNA. The lack of sequenced organelle genomes of green algae prevented the comparison of genomic architectures in other closely related species. In this study, I expanded on the information from the 2010 …

Planting The Chalcone Reductase Family Tree: Identification And Characterization Of Chalcone Reductase Genes In Soybean, Caroline Julia Sepiol

Electronic Thesis and Dissertation Repository

Soybean (Glycine max [L.] Merr) is an important crop grown in Canada, generating $2.4 billion in sales. Though this number may be promising, soybean farmers lose about $50 million worth of yield annually due to root and stem rot disease caused by Phytophthora sojae. Many strategies have been developed to combat the infection; however, these methods are prohibitively expensive. A ‘cost effective’ approach to this problem is to select a trait naturally found in soybean that can increase resistance. One such trait is the increased production of root glyceollins. One of the key enzymes exclusively involved in glyceollin …

Genetic Analysis Of A Non-Germinating Mutant Of Arabidopsis Thaliana, Md Jakir Hossan

Electronic Thesis and Dissertation Repository

Seed germination is partially controlled by plant hormone gibberellins (GAs). Chemical mutagenesis yielded an Arabidopsis thaliana mutant gm11, which has an absolute gibberellin requirement for seed germination. This mutant exhibited phenotypes of GA-rescuable dwarfs, including dark-green leaves, and reduced fertility. However, with repeated GA treatment, gm11 develops into fertile plants with a nearly wild type phenotype. Bulked-segregant analysis mapped gm11 to the bottom arm of chromosome 1, and subsequent next-generation mapping revealed that the mutation is a G → A transition in At1g79460 (GA2), creating a premature stop codon. This gene encodes an ent-kaurene synthase (KS) which catalyzes …

Determination Of The Phylogenetic Range Of Sex Combs Reduced Activity In Drosophila Melanogaster, Laura E. Garofalo

Electronic Thesis and Dissertation Repository

The homeotic selector (Hox) genes are required for body patterning in bilaterians. Sex combs reduced (SCR) is a HOX protein in Drosophila melanogaster with two activities: SCR^T1 and SCR^lab activity required for patterning the prothorax (T1) and labia, respectively. SCR^T1 is proposed to be conserved throughout bilaterians while the phylogenetic range of functional conservation of SCR^lab is comparatively unknown. The goal of this work was to elucidate the evolutionary time point at which SCR activity changed. CRISPR/Cas9 transgenesis was used to incorporate ɸC31 integrase recombination sites in Drosophila Scr. The ɸC31 integrase could …

Impact Of Estrogen Receptor Alpha On Sle1-Induced Loss Of Tolerance, Shayla D. Yoachim

Theses & Dissertations

The autoimmune disease lupus shows a significant female sex bias. This sex bias may be due to the ability of estrogens to promote loss of tolerance to chromatin, the initial loss of tolerance event in lupus. Previously, we demonstrated that the ability of estrogens to promote lupus in (NZBxNZW) F1 mice is dependent on signaling via estrogen receptor alpha (ERα). The Sle1 lupus susceptibility allele controls loss of tolerance to chromatin, and C57BL/6 (B6) mice carrying the Sle1 lose tolerance and develop anti-chromatin autoantibodies and spontaneously activated immune cells. Loss of tolerance occurs earlier and with a higher penetrance in …

Role Of The C-Terminus Of The Catalytic Subunit Of Translesion Synthesis Polymerase Ζ (Zeta) In Uv-Induced Mutagensis, Hollie M. Siebler

Theses & Dissertations

Cellular DNA is under constant attack by endogenous and exogenous DNA damaging agents that threaten genome integrity. Unrepaired DNA lesions often stall replicative DNA polymerases and are bypassed by translesion synthesis (TLS) to prevent replication fork collapse. TLS mechanisms are lesion- and species-specific, with prominent roles of specialized DNA polymerases with relaxed active sites. After incorporation of nucleotide(s) across from the lesion, the distorted primer termini are typically extended by DNA polymerase ζ (Pol ζ). As a result, Pol ζ is responsible for most DNA damage-induced mutations. Mechanisms of sequential polymerase switches and regulation of Pol ζ access to DNA …

Functional Characterization Of The Roles Of Endocytic Recycling Regulator Ehd1 Using In Vivo And In Vitro Analyses, Priyanka Arya

Theses & Dissertations

Endocytic recycling is a fundamental cellular process that allows the precise regulation of the membrane components and receptors at the cell surface. Recent studies have established that the C-terminal Eps15 homology domain-containing (EHD) proteins function as key regulators of this process. Four highly-conserved members of the EHD protein family in mammals, EHD1-EHD4, play shared as well as unique roles in endocytic trafficking. Studies presented here demonstrate a critical role of EHD1 in the normal ocular development in mice. Ehd1 knockout mice generated in our laboratory displayed gross ocular phenotypes including the anophthalmia, microphthalmia, and congenital cataracts. Hematoxylin and eosin (H&E) …

Genetic Modification Of Switchgrass (Panicum Virgatum L.) For Improvement Of Plant Architecture, Biomass Productivity And Sugar Release Efficiency For Biofuel, Wegi Aberra Wuddineh

Doctoral Dissertations

Switchgrass (Panicum virgatum L.) is a leading candidate bioenergy crop for sustainable biofuel production. To ensure its economic viability, tremendous improvements in switchgrass biomass productivity and recalcitrance to enzymatic saccharification are needed. Genetic manipulation of lignin biosynthesis by targeting transcriptional regulators of higher level domains of lignin biosynthesis and other complex traits could alter several bioenergy-desirable traits at once. A three-pronged approach was made in the dissertation research to target one plant growth regulator and transcription factors to alter plant architecture and cell wall biosynthesis.

Gibberellin (GA) catabolic enzymes, GA 2-oxidases (GA2oxs), were utilized to alternatively modify the lignin …

Evolution And Phylogeny Of The Parasitoid Subfamily Phasiinae (Diptera: Tachinidae), Jeremy Daniel Blaschke

Doctoral Dissertations

The first molecular phylogenetic analysis of the agriculturally important parasitoid subfamily Phasiinae (Diptera: Tachinidae) is presented, estimated from 128 worldwide taxa (80 genera) and approximately 7.6 kilobases of nuclear data. Special emphasis is placed on taxa with controversial taxonomic placement. The resultant phylogenetic tree is used to reconstruct ancestral character states, trace the evolution of significant adaptive traits within the Tachinidae, and test hypotheses about the classification of Phasiinae. Subfamily placements of the taxa Eutherini, Epigrimyiini, Litophasia, Strongygastrini, and Parerigonini are confidently resolved, the former three within Dexiinae and the latter two within Phasiinae. Due to sparse molecular evidence, …

Calmodulin-Like Protein 38: A Component Of Ribonucleoprotein Particles During Hypoxic Stress Responses In Arabidopsis, Ansul Lokdarshi

Doctoral Dissertations

Waterlogging stress leads to a crisis in energy metabolism and the accumulation of toxic metabolites due to the hypoxic and/or anoxic environment associated with this condition. To respond and adapt to this situation, higher plants employ an integrated genetic program that leads to the induction of anaerobic response polypeptide genes that encode metabolic and signaling proteins involved in altering metabolic flow and other adaptive responses. The study presented here shows that the Arabidopsis thaliana calmodulin-like protein CML38 is calcium sensor protein that serves as a member of the core anaerobic response gene family and is involved in modulating the survival …

Distinct Physiological Roles For The Two Isoforms Of The Er Chaperone Grp170 In Caenorhabditis Elegans, Yuanyuan Li

Biology Theses

GRP170 is a large molecular chaperone found in the ER of all eukaryotes. The nematode Caenorhabditis elegans has two loci encoding GRP170: T24H7.2 (grp170a) and T14G8.3 (grp170b). The phenotypes of nematodes genetically deficient for either grp170a or grp170b were compared to a standard laboratory strain with functional grp170 loci. Worms that were deficient for grp170a developed 32% slower than the control strain. The loss of grp170a had a significant but modest reduction on the life span compared to the control strain. Worms deficient for grp170a also displayed significantly increased embryonic lethality and resulted in 6.9% arrested embryos. The loss of …

Differential Regulation Of The Two Grp170 Paralogues Of Caenorhabditis Elegans, Antonio L. Rockwell

Biology Theses

Abstract of a Thesis

Differential Regulation of the Two grp170 Paralogues of Caenorhabditis elegans

Caenorhabditis elegans has two loci encoding the large eukaryotic molecular chaperone Grp170, grp170a (T24H7.2) and grp170b (T14G8.3). To investigate expression of the two C. elegans grp170 loci during ER stress, the Unfolded Protein Response (UPR) was induced with the glycosylation inhibitor tunicamycin. Levels of grp170a mRNA did not significantly change in response to tunicamycin treatment while the levels of grp170b mRNA increased 6-fold. ER stress induction of grp170b was unaffected in worms defective for the ATF6 and PERK-1 UPR signal transduction pathways. However, worms defective …

Germline Mutation Detection In Next Generation Sequencing Data And Tp53 Mutation Carrier Probability Estimation For Li-Fraumeni Syndrome, Gang Peng

Dissertations & Theses (Open Access)

Next generation sequencing technology has been widely used in genomic analysis, but its application has been compromised by the missing true variants, especially when these variants are rare. We proposed a family-based variant calling method, FamSeq, integrating Mendelian transmission information with de novo mutation and sequencing data to improve the variant calling accuracy. We investigated the factors impacting the improvement of family-based variant calling in simulation data and validated it in real sequencing data. In both simulation and real data, FamSeq works better than the single individual based method.

In FamSeq, we implemented four different methods for the Mendelian genetic …

Detection Of Genes Influencing Chronic And Mendelian Disease Via Loss-Of-Function Variation, Alexander H. Li

Dissertations & Theses (Open Access)

A typical human exome harbors dozens of loss-of-function (LOF) variants predicted to severely disrupt or abolish gene function. These variants are enriched at the extremely rare end of the allele frequency spectrum (< 0.1%), suggesting purifying selection against these sites. However, most previous population-based sequencing studies have not included analysis of genotype-phenotype relationships with LOF variants. Thus, the contribution of LOF variation to health and disease within the general population remains largely uncharacterized.

Using whole exome sequence from 8,554 participants in the Atherosclerosis Risk in Communities (ARIC) study, we explored the impact of LOF variation on a broad spectrum of human phenotypes. First, we selected 20 common chronic disease risk factor phenotypes and performed gene-based association tests. Analysis of this sample verified two relationships in well-studied genes (PCSK9 and APOC3) and identified eight new loci. Novel relationships included …

Hippocalcin Response To Calcium: Do Conserved Tryptophans – W30 Or W103 – Matter?, Sunkesula K. Sagar

Graduate School of Biomedical Sciences Theses and Dissertations

Changes in intracellular calcium levels play a very important role in cell signaling, in turn, affecting neuronal functions such as memory, learning and cell death. A class of proteins called Neuronal Calcium Sensor (NCS) proteins serves to modulate the functioning of the neuronal cells in response to changes in calcium levels, and prevent neuronal apoptosis. Structurally, all NCS proteins have 4 calcium-binding EF hand motifs, although EF1 does not bind to calcium in many members. All NCS proteins have an acyl modification at the N- terminus – where a myristoyl group is added post-translationally. Hippocalcin (HPCA) is an NCS protein, …

Functional Analysis Of Synthetic Gene Circuits Controlling A Protein Pump In Yeast, Junchen Diao

Dissertations & Theses (Open Access)

Synthetic biology aims to build biological devices to understand living systems and explore new applications. Synthetic gene circuits such as genetic switches, oscillators and logic gates are at the core of many synthetic biology applications. These gene circuits often include a sensor/regulator protein capable to detect small molecules and then transduce them into a regulatory signal to generate measurable output. Similar signal transduction networks are also abundant in nature. However, in many natural and engineered scenarios, the output also affects the regulator/sensor protein. How such interactions between the regulator/sensor and the output affect synthetic gene circuit function has not been …

Population Structure Of Late Blight (Phytophthora Infestans) In Colombia And Ecuador And Downy Mildew (Peronospora Farinosa F. Sp. Spinaciae) On Spinach In Arizona And California, Rebecca Marie Lyon

Masters Theses

In this study, Phytophthora infestans and Peronospora farinosa f. sp. spinaciae populations were analyzed using single nucleotide polymorphisms (SNPs). In Ecuador and Colombia, Phytophthora infestans causes significant damage to potato and tomato and the epidemiology is known to be highly clonal. Our objective was to measure population structure within the context of this clonal epidemiology using both synonymous and nonsynonymous markers. Candidate SNP sites were selected by comparing the draft genomes of the Ecuadorian isolates EC1-3527 and EC1-3626. Genotypes were assessed directly from infected tissue using a targeted sequencing approach. A total of 54 polymorphic sites were assessed in 93 …

An Analysis Of The Differential Methylation And Expression Of Imprinted Genes In M. M. Musculus, M. M. Domesticus, And Their Hybrids, Anna P. Rice

All NMU Master's Theses

Epigenetics has been found to have an effect on many aspects of biology. Epigenetics refers to modifications of the double-stranded DNA molecule, which do not change the nucleotide sequence but do affect gene expression. DNA methylation is a type of epigenetic modification. Genomic imprinting is a pattern of gene expression that is primarily achieved through DNA methylation, and it results in the expression of only one allele at a particular locus. In this study, I analyzed the methylation patterns of five imprinted genes in the hybrids of two different lab strains of the house mouse subspecies, M. m. musculus and …

Identification Of Familial Wilms Tumor Predisposition Genes Using Whole Genome Sequencing, Timothy B. Palculict

Dissertations & Theses (Open Access)

Wilms tumor, a childhood tumor arising from undifferentiated renal mesenchyme, is diagnosed in North America at a frequency of 1 in 10,000 live births and accounts for 5% of all pediatric cancers. The etiology of Wilms tumor is heterogeneous with multiple genes known to have an effect on Wilms tumor development; however, these genes are rarely associated with familial Wilms tumor. Gene mutations in WT1, WTX, CTNNB1 and TP53 are observed in a third of sporadic tumors, while the causative gene(s) responsible for familial Wilms tumor are largely unknown. Approximately 2% of Wilms tumor patients have a family …

In Vivo Significance Of The Mdm4 And P73 Interaction During Development And Tumorigenesis, Mehrnoosh Tashakori

Dissertations & Theses (Open Access)

The tumor suppressor protein p53 is negatively regulated by Mdm4 protein. The significance of such regulation was determined from mouse models. Mdm4-deficient mice are embryonic lethal at E7.5 in a p53-dependent manner. p73, a member of the p53-family, is a transcription factor with tumor suppressor activity. In vitro studies show that Mdm4 binds to p73 and, further, comprehensive biochemical studies revealed that Mdm4 has higher affinity for p73 than p53. However, little is known about the significance of the Mdm4 and p73 interaction in vivo. This study aimed to elucidate the biological consequences of this interaction during embryogenesis …

Clinical And Mechanistic Insights Into Novel Probiotic Functions And Formulations, Jordan Bisanz

Electronic Thesis and Dissertation Repository

Using a combination of hypothesis and discovery based approaches, the goal of this thesis was to better describe novel probiotic functions and their mechanisms while striving to better understand the effect of formulation on Bifidobacterium animalis subsp. lactis, Lactobacillus paracasei and L. rhamnosus.

Using RNA-Seq, a bacterial metatranscriptome analysis of a commonly consumed probiotic yogurt showed that the organisms adapted to storage time and flavor additions. This led to the discovery that in addition to the probiotic health benefits, members of the L. casei group (L. rhamnosus and L. paracasei) produce volatile sulfur compounds mediated by a novel …

Assessment Of Genome-Wide Genetic And Epigenetic De Novo Variation In Families With Monozygotic Twins Discordant For Schizophrenia, Christina A. Castellani

Electronic Thesis and Dissertation Repository

Schizophrenia (OMIM: 181500) is a common, debilitating and life-altering disorder. It affects 1% of the population worldwide and most often presents in early adulthood leading to devastating effects for patients, their families and society. Despite thousands of studies performed on the underlying mechanisms of schizophrenia, the causes of the disease remain unknown. However, what is known is that environmental, genetic and epigenetic factors contribute to the development of this complex disorder. Although a genetic role in schizophrenia is well established, the search for schizophrenia genes using traditional approaches has remained challenging. Interestingly, monozygotic twins show concordance for schizophrenia only 50% …

Experimental Test Of Genetic Rescue In Isolated Populations Of Brook Trout, Zachary L. Robinson

Masters Theses

Translocations are an important aspect of the management of natural populations in an increasingly fragmented landscape. Maintaining connectivity and gene flow is beneficial for both contemporary fitness and adaptive potential in the face of environmental change. Genetic rescue (GR) can alleviate inbreeding depression, genetic load, and increase adaptive potential of populations. Here, I have translocated 10 (5 of each sex) brook trout (Salvelinus fontinalis) to four geographically proximate and environmentally similar fragmented stream-dwelling populations of brook trout in Virginia to test for genetic rescue. The translocated brook trout contributed to more families than would be expected under neutral …

Population Structure Of Central Stoneroller (Campostoma Anomalum) On The Ozark Plateau In Arkansas And Missouri, Mallory Jane Jeffers

Graduate Theses and Dissertations

Stream fishes on the Ozark Plateau have been influenced both by historic events (i.e. vicariance versus dispersal) and ecological forces (i.e. flow regime). To examine the role of these processes, genetic structure of Central Stoneroller (Campostoma anomalum), an ecologically important omnivorous minnow with a broad distribution and elevated abundance, was evaluated across populations in the White River drainage of the Ozark Plateau in Arkansas and Missouri. Fin clips of five to eight individuals were taken at each of 20 sites (N=138 individuals; average=6.9), selected so as to represent two different flow regimes: intermittently flashy (N=10 sites; N=73 individuals; average=7.3/ site) …

Genetic Variation And Species Distribution Of Subterranean Termites In The Southeastern U.S. Focusing On Family Structure And The Invasive Formosan Subterranean Termite, Mark Allan Janowiecki

Graduate Theses and Dissertations

This work applies molecular genetic tools to distinguish the identity and understand the biology of termites, particularly Reticulitermes Holmgren and Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) in the southeastern U.S. Termites are important economic pests costing billions of dollars annually to Americans but also are important wood decomposers in natural settings. Molecular tools are essential for species identification because of the morphological ambiguities among species. The cryptic nest structure of subterranean termites which prevents adequate sampling makes molecular genetic tools essential to examine an entire colony.

A molecular diagnostic technique was created to differentiate Coptotermes formosanus, an invasive species in the …

Cell Wall Mutants In Arabidopsis Thaliana, Christy Jane Moore

Theses and Dissertations

Plant cell walls are versatile structures, playing important roles in communication, defense, organization and support. The importance of each of these functions varies by cell type, with specialized cells often utilizing one or two functions more than others. Trichomes, or leaf hairs, and hypocotyl cells for instance, exhibit distinct cell wall characteristics. Trichomes have developed very thick cell walls with several raised structures, known as papillae, on their surfaces. It is believed that these cells function in defense against predators, making it difficult to crawl on the leaf surface, and in protection against ultra violet radiation, through refraction of light …

The Effects Of Sustained, High-Velocity Exercise On Gene Expression In California Yellowtail (Seriola Lalandi), Kelli Hatter

Undergraduate Honors Theses

California Yellowtail muscle fibers have been observed to exhibit two drastically different development patterns resulting from the speeds at which they are exercised. When fish are exercised at a moderate rate their epaxial fast-twitch muscle fibers grow in diameter—hypertrophy; when they are exercised at a fast speed, more new epaxial fast-twitch muscle fibers are produced—hyperplasia. To determine the underlying reason for this difference in muscle development, my summer research project and honors thesis exercised fish at: fast, moderate, and control speeds for a sustained amount of time to determine what is happening on a cellular level to cause the observed …

Investigating The Role Of Wolbachia Endosymbionts In The Expansion Of The F Element In Drosophila Ananassae, Elizabeth J. Chen

Undergraduate Theses—Unrestricted

At 4.2 Mb overall, the Drosophila melanogaster Muller F element (dot chromosome) is an unusual autosome; it is broadly heterochromatic, but the distal 1.3 Mb has a gene density and expression pattern similar to other autosomes. More intriguing is the large expansion of the D. ananassae F element (~20 Mb). Elucidating the factors that contribute to this expansion could improve our understanding of how heterochromatic domains are maintained and amplified.

Previous analyses show that the lateral gene transfer (LGT) of Wolbachia (the most widespread intracellular bacteria in the Rickettsiales order) into the D. ananassae genome is an important contributor to …

Addressing The Black Box Phenomenon Of Genome Sequencing And Assembly, Brandon Carter

Senior Honors Projects, 2010-2019

Genomics, a study of all genetic material in an organism, is a new discipline having a great impact on medicine, agriculture, and environmental phenomena. Most undergraduate faculty members were not formally trained in genomics and must retool themselves in order to stay current with these evolving technologies. Advances in sequencing technology have resulted in an explosion of “big data” that can only be managed and analyzed using digital methods. Multiple complex computer programs are required to teach students the concepts using hands-on methods. These programs are challenging to use, especially since the same faculty members lacking genomics training were not …

Divergence Of Vertebrate And Insect Specific Toxin Genes Between Three Species Of Widow Spider, Shannon G. Kahn

Honors College Theses

The Brown widow spider, Latrodectus geometricus, is an introduced species to the southern United States (Brown 2008). The Brown widow is a member of the widow spider genus Latrodectus which includes the southern Black widow (L. mactans) and 29 other venomous species. All species of widow spiders produce venom which is used against both predator and prey. These venoms are composed of several different species-specific toxins, each encoded by a different gene (Graudins 2012). Previous research has shown that positive selection pressures affect the venom of snakes and snails, thus aiding in adaptive potential of the species …