Life Sciences Commons^™

Chemosensory Receptors In Berghia Stephanieae: Bioinformatics And Localization, Kelsi L. Watkins

Masters Theses

Chemosensation is achieved through the binding of chemical signals to chemoreceptor proteins embedded in the membranes of sensory neurons. The molecular identity of these receptors, as well as the downstream processing of chemosensory signals, has been well studied in arthropods and vertebrates. However, very little is known about molluscan chemosensation. The identity of chemoreceptor proteins in the nudibranch mollusc Berghia stephanieae are unknown. Data from other protostome and molluscan studies suggest Berghia may use ionotropic receptors for some forms of chemoreception. This study used a bioinformatics approach to identify potential chemosensory ionotropic receptors in the transcriptome of Berghia. A …

Cloning Of Root-Specific Promoters In Helianthus Annuus And Populus Trichocarpa, Jacob Murphy

Masters Theses

Tissue-specific promoters are a valuable tool in plant biotechnology because they allow for precise spatial control of transgene expression. To identify root-specific promoters in sunflower (Helianthus annuus), sunflower RNA sequence (RNA-seq) data were screened in silico by Pavlidis Template Matching (PTM) with a root-specific template. Eight root-specific genes were identified in sunflower. The 2.5kbp promoter regions of six of these root-specific sunflower genes (HaTOR1L3, HaGOXL3A, HaGOXL3B, HaAPA1, HaPOE1, and HaMOP10) were successfully cloned as well as the 3kbp promoter regions of four root-specific genes previously identified in poplar (Populus trichocarpa …

Molecular Markers Within And Beyond Viburnum: A Single Set Of Ssrs For An Immense Genus, Trinity Paige Hamm

Masters Theses

Comprised of more than 160 species, Viburnum is the largest genus in the Viburnaceae (formerly Adoxaceae) alongside Adoxa and Sambucus. The native range of Viburnum species spans much of the Northern Hemisphere and extends into the mountains of South America and Southeastern Asia. Their wide geographic range has made Viburnum particularly interesting to phylogeographers and phylogeneticists. This diverse genus is also horticulturally valuable with varying traits such as fragrance, fruit color, and bud and inflorescence form. There is sufficient morphological diversity in the genus for there to be more than 70 species and intraspecific hybrids in cultivation, and in …

Structure-Function Studies Of The Trypanosome Mitochondrial Replication Protein Polib, Raveen Armstrong

Masters Theses

Trypanosoma brucei and related protists are distinguished from all other eukaryotes by an unusual mitochondrial genome known as kinetoplast DNA (kDNA) that is a catenated network composed of minicircles and maxicircles. Replication of this single nucleoid involves a release, replicate, and reattach mechanism for the thousands of catenated minicircles and requires at least three DNA polymerase (POLIB, POLIC and POLID) with similarity to E. coli DNA polymerase I. Like other proofreading replicative DNA polymerases, POLIB has both an annotated polymerase domain and an exonuclease domain. Predictive modelling of POLIB indicates that it has the canonical right hand …

Pedigree And Molecular Assessment Of Relatedness In A Captive Chimpanzee Population, Francesca Golus

Masters Theses

As a crucial component of biodiversity, genetic diversity contributes to variability among individuals, allowing populations of endangered species to be resilient in the face of changing environmental conditions. Zoological institutions have become a cornerstone of conservation efforts and a refuge for endangered species given threats imposed on wild populations by climate change, habitat fragmentation and degradation, and overexploitation. Chimpanzees (Pan troglodytes) are an endangered species experiencing drastic population decline in the wild, yet are common residents in zoos and wildlife sanctuaries. To sustain long-term, genetically viable populations of species of concern, zoos use studbooks and paternity testing to …

Genome-Wide Characterization Of Lateral Organ Boundaries Domain Genes In Sunflower (Helianthus Annuus L.), Saleh M. Aljurbua

Masters Theses

Lateral Organ Boundaries (LBD) gene family encodes transcription factors found only in plants with essential roles in growth, development, and response to biotic and abiotic factors. The sunflower (Helianthus annuus L., Asteraceae family) LBD genes were studied using different in silico and phylogenetic approaches including four major steps: gene identification, phylogenic analysis, structural analysis, and expression analysis. Sixty-four sunflower LBD genes (HaLBDs) were identified using query search of all Arabidopsis LBD genes against sunflower genome (version r1.2) followed by LBD domain sequence verification. A phylogenetic tree was reconstructed using MEGA-X program. The identified genes were phylogenetically classified into two main …

Carbon Metabolism In Cave Subaerial Biofilms, Victoria E. Frazier

Masters Theses

Subaerial biofilms (SABs) grow at the interface between the atmosphere and rock surfaces in terrestrial and subterranean environments around the world. Multi-colored SABs colonizing relatively dry and nutrient-limited cave surfaces are known to contain microbes putatively involved in chemolithoautotrophic processes using inorganic carbon like carbon dioxide (CO₂) or methane (CH₄). However, the importance of CO₂ and CH₄ to SAB biomass production has not been quantified, the environmental conditions influencing biomass production and diversity have not been thoroughly evaluated, and stable carbon and nitrogen isotope compositions have yet to be determined from epigenic cave SABs. …

The Transcriptome Response Of The White-Rot Fungus Trametes Versicolor To Wild-Type And Lignin-Modified Hybrid Poplar, Anbarah Alzabaidi

Masters Theses

Plant biomass is a renewable and sustainable feedstock for biofuel production that can reduce societal dependence on fossil fuels. However, the production of liquid biofuels from the non-starch (i.e. lignocellulosic) material through fermentation technology is limited due to the complexity of the cell wall structure. This necessitates the use of chemical, thermal, and/or mechanical pretreatment technologies, which adds significant capital, operational, and environmental costs. Biological pretreatment strategies have the potential to mitigate these expenses by harnessing the innate ability of specialized bacteria and fungi to deconstruct lignocellulose. White-rot fungi (e.g. Trametes versicolor) have been shown to be …

Detection Of Beta-Lactamase Variants In Municipal Wastewater And Fresh Water, Sunil Pandey

Masters Theses

The occurrence and spread of antibiotic-resistant genes (ARGs) are pressing public health problems worldwide. A key factor contributing to the spread of ARGs is lateral gene transfer. Wastewater treatment plants (WWTPs) are measured hot spots of microbial diversity and resistance because they receive polluted wastewater from diverse sources and contain a variety of different environments with dense bacterial loads. Due to the overuse of antibiotics the genetic capacities of microbes have profited. This helps every source of resistance gene and every means of horizontal gene transmission to develop the multiple mechanism of resistance to each antibiotic used clinically, agriculturally, or …

Stressing Out: Dynamics Of Chromatin Insulator Body Formation In Eukaryotes Under Osmotic Stress, Shannon Marie Garland

Masters Theses

Higher order genome organization and the role it plays in governing cell dynamics and protein expression has become a widely studied field. Chromatin insulators are important to this organization in their ability to form long range contacts between distant regions on the genome. During times of osmotic stress, insulator proteins leave their binding sites on the DNA to form insulator bodies in the nucleus. This phenomenon relieves the DNA of its structure and is rapidly reversible. Using a variety of immunofluorescent staining methods, this work looked to further characterize this process while attempting to identify molecular mechanisms that mediate chromatin …

Detection Of Antibiotic Resistance Loci In Wastewater And Fresh Water Samples, Mohammed Hamed Almalki

Masters Theses

Proper treatment of municipal wastewater is important not only for prevention of eutrophication but also for public health concerns. One consideration for wastewater treatment is the presence of organisms carrying antibiotic resistance genes. If these organisms survive the wastewater treatment process and are released into the streams, they can pose significant risks to public health. In addition, their presence also raises concerns for lateral gene transfer events at the wastewater plant. The spread of antibiotic loci has a major impact in public health and clinical therapy because resistant organisms render conventional treatments of important illnesses difficult. Beta-lactam drugs are known …

A Population Genetics Study Of Discula Destructiva, The Causal Agent Of Dogwood Anthracnose, Kristie Lynn Mantooth

Masters Theses

Flowering dogwood (Cornus florida L.) is prized as an ornamental and contributes millions of dollars to the economy through tourism and sales each year. Dogwood anthracnose, caused by Discula destructiva Redlin, was observed in the late 1970s on the east and west coasts of the United States and by 1991 had quickly spread throughout much of the native ranges of C. florida and C. nuttalli. The objectives of this study were to investigate the genetic diversity and population structure of D. destructiva using simple sequence repeats (SSRs), and to test the cross-transferability of these markers to other Discula …

Population Genetics And Genomics Within The Genus Pityopsis, Elizabeth Anne Hatmaker

Masters Theses

Pityopsis (Asteraceae) includes seven species; one species, P. ruthii, is federally endangered. The genus exhibits a range of ploidy levels, widespread hybridization among species with overlapping ranges, and interesting adaptive traits such as fire-stimulated flowering. However, taxonomy of Pityopsis has remained unresolved. Resolving interspecific relationships can lead to a deeper understanding of the inheritance and hybridization patterns, as well as the evolution of adaptable traits. Our first objective was to examine population structure and gene flow within Pityopsis ruthii. Polymorphic microsatellite markers (7 chloroplast and 12 nuclear) were developed and used to examine genetic diversity of 814 P. …

Impact Of Transcription Factors On Fibroblast Specific Gene Silencing In Somatic Cell Hybrids, Tania Ray

Masters Theses

As totipotent cells differentiate to specific cell types in multicellular organisms, certain sets of genes are turned off (gene silencing/gene extinction) while others are turned on (gene activation). Cell type specific gene silencing and gene activation is the basis of cell differentiation. Somatic cell hybrids which are produced by fusing different types of somatic cells from two different tissues have been used as a valuable resource to discern the phenomenon of lineage- specific gene extinction. Identification of regulatory factors that mediate gene extinction in hybrids are key to understanding regulatory mechanisms that govern cell differentiation in a multicellular organism. Transcription …

Rare Occurrences Of Free-Living Bacteria Belonging To Sedimenticola From Subtidal Seagrass Beds Associated With The Lucinid Clam, Stewartia Floridana, Aaron M. Goemann

Masters Theses

Lucinid clams and their sulfur-oxidizing endosymbionts comprise two compartments of a three-stage, biogeochemical relationship among the clams, seagrasses, and microbial communities in marine sediments. A population of the lucinid clam, Stewartia floridana, was sampled from a subtidal seagrass bed at Bokeelia Island Seaport in Florida to test the hypotheses: (1) S. floridana, like other lucinids, are more abundant in seagrass beds than bare sediments; (2) S. floridana gill microbiomes are dominated by one bacterial operational taxonomic unit (OTU) at a sequence similarity threshold level of 97% (a common cutoff for species level taxonomy) from 16S rRNA genes; …

Sequence Analysis Of Maize Yellow Stripe3 Candidate Genes, Dennis B. Depaolo

Masters Theses

The work presented here focuses on the molecular mechanism of phytosiderophore secretion in graminaceous plants. In maize, yellow stripe3 (ys3) is a mutant that is deficient in its ability to secrete iron-chelating compounds of the mugineic acid family known as phytosiderophores. Phytosiderophores are specific to grasses and are used for the acquisition of iron. Genetic linkage mapping of the ys3 locus lead to a region of interest on chromosome 3 defined by marker UMC1773. The sequence of eleven candidate genes (GRMZM2G390345, GRMZM2G390374, GRMZM2G342821, GRMZM5G800764, GRMZM2G502560, GRMZM5G849435, GRMZM2G105766, GRMZM5G876835, GRMZM2G036976, GRMZM2G502563, miR167g) revealed several small deletions …

Rapid Method Of Processing Sperm For Nucleic Acid Extraction In Clinical Research, Matthew K. De Gannes

Masters Theses

Background: Sperm contain highly compact nuclei, inhibiting DNA extraction using traditional techniques. Current methods extracting sperm DNA involve lengthy lysis and no means of stabilizing DNA, hindering clinical research.

Objective: We sought to optimize an efficient method of extracting high quality human sperm DNA.

Methods: Sperm from three volunteers were isolated using PureCeption. We tested 1) proteinase K with DNA/RNA Shield, 2) DTT and TCEP as reducing agents, 3) QIAshredder homogenization, and 4) stability of sperm DNA fresh (baseline) or after 4 weeks of storage at 4^OC in DNA/RNA Shield using modified Quick-gDNA MiniPrep. DNA was PCR amplified …

Molecular Marker Assisted Backcross Development And Evaluation Of An Environmentally Friendly, Commercially Acceptable Low Seed Phytate Soybean, Jeffrey David Boehm Jr.

Masters Theses

Soybean [Glycine max (L). Merrill] is the world’s top oilseed crop. High protein soymeal is a primary soybean use and staple food of livestock and fish. The storage form of phosphorus (P) in the seed is phytic acid, or phytate [myo-inositol 1,2,3,4,5,6 hexakisphosphate] which binds and chelates key nutritional cations forming stable salts that are non-digestible. Livestock waste laden with phytate P is an environmental concern, and a source of nonpoint P pollution causing eutrophication and surface water quality deterioration. The primary objective of this research was the development of a commercially acceptable low phytate (LP) soybean …

Molecular Systematics Of The Subfamily Phasiinae (Diptera: Tachinidae), Jeremy Daniel Blaschke

Masters Theses

The subfamily Phasiinae (Diptera: Tachinidae) is composed of obligate endoparasitoid flies that attack heteropteran insects, many of which are important agricultural pests. To lay a foundation for future studies in phasiine life history and their potential for use as biological control agents, the phylogenetic relationships within Phasiinae were explored using nucleotide sequences of two genes, CAD and LGL. A total of 63 taxa for CAD and 73 taxa for LGL were included in the analyses, representing 58 genera. Maximum likelihood inference methods were employed to reconstruct phylogenetic relationships in separate analyses of each gene and in a combined dataset. Phylogenetic …

Trends In Borrelia Spp. Prevalence In Ixodes Spp. Ticks From The Southeastern Coastal United States, Lauren Paul Maestas

Masters Theses

The Lyme borreliosis (LB) cycle, involving Ixodes scapularis and Borrelia burgdorferi sensu stricto (Bbss), is well documented in the northeastern US, where LB is becoming increasingly prevalent. In coastal North Carolina, I. affinis has been shown to have a higher incidence of Bbss than I. scapularis. My objectives were, to assess changes in prevalence of Bbss in Ixodes spp. along a transect from Virginia to Florida, and to assess the value of dogs and mesomammals as sentinels for spread of Borrelia burgdorferi sensu lato.

Ixodes spp. were collected at sites from 37.4^o N to 30.0^{o …}

Genomic Analysis And Functional Characterization Of The Terpene Synthase Gene Family In Brachypodium Distachyon, Ayla Marie Norris

Masters Theses

Brachypodium distachyon is a small, temperate grass native to the Mediterranean region. While not agriculturally important, it possesses many characteristics such as rapid generation time, self-pollination, a small genome size, and ease of genetic transformation, that make it an attractive model organism for genetic studies. Brachypodium is closely related to other grasses of economical importance such as rice, sorghum, and especially wheat, and in fact it is the first member of the Pooideae grass sub-family to have its genome fully sequenced. Cereal grasses such as rice, sorghum, and maize have been reported to produce a class of secondary metabolites called …

Microarray Analysis Of Mrna Translation State In Arabidopsis Thaliana, Bayu Sisay Tiruneh

Masters Theses

The synthesis of proteins – translation – is one of the cellular processes that consume a great deal of energy. Genome-wide assays performed in Arabidopsis and other organisms have revealed that the translation status of mRNAs responds dramatically to different stresses and environmental and growth signals. Similar assays have revealed how translation of specific mRNAs is perturbed by genetic mutations in eukaryotic translation initiation factors (eIFs). I performed genome-wide analyses of translation state in A. thaliana seedlings with mutant genotypes for a specific eIF (subunit h of eIF3) and the large ribosomal protein, L24 (RPL24B). Using data from prior microarray …

Functional Analysis Of The Cyp6a8 Gene Promoter Of Drosophila Melanogaster For Caffeine- And Phenobarbital-Inducibility By Site-Directed Mutagenesis, Olivia Nichole Hill

Masters Theses

Cytochrome P450 enzymes (CYPs), found in almost all organisms, are involved in endobiotic metabolism and detoxification of xenobiotic compounds, such as drugs, pollutants, and insecticides. In insects, CYPs play a major role in conferring resistance to various insecticides including DDT. In Drosophila and other insects, DDT-resistant strains exhibit increased expression of multiple P450 genes; however, the mechanism of overexpression is unknown. Since many CYP genes including Cyp6a8 of Drosophila are induced by caffeine and other xenobiotics, these chemicals are used as tools to understand the regulation of these genes. Previously it was shown that the 0.8-kb (-1/-732) and 0.2-kb (-1/-170) …

Mitochondrial Dna And Microsatellite Genetic Variation Of Dollar Sunfish (Lepomis Marginatus), Nancy A. Schable

Masters Theses

Dollar sunfish (Lepomis marginatus) at the Savannah River Site were sampled from two radiocesium contaminated ponds with a history of thermal elevation and from reference ponds with no history of radioactive or thermal contamination. Fluorescent sequencing of a portion of the mitochondrial control region showed no genetic variation within or between populations. Centrarchidae microsatellite loci Lma 120 (6 alleles, N=124 from 5 populations), Lma 20 (10 alleles, N=37 from 3 populations) and Rb 7 (14 alleles. N=17 from 2 populations) were variable for dollar sunfish. An additional 9 microsatellite primer pairs were designed for dollar sunfish. Testing of the designed …

Partial Characterization Of Lpss From Rhizobium Fredii Usda 205 Before And After Nod Gene Induction, Bradley Lawrence Reuhs

Masters Theses

The lipopolysaccharides (LPS) from Rhizobium fredii USDA 205 and its nod- mutant, HC 205, which has been cured of the 192-MDa symbiotic plasmid, were isolated by the phenol-water extraction method. The LPS from the wild-type strain which was grown in association with host-plant root extract or a flavone analog to a constituent of the root extract, which is required for the induction of at least a portion of the nodulation genes, was also isolated. The conditions required for the induction of the nod genes were determined by work with a nod- mutant of R. fredii USDA 201 which …

A Comparison Of The Acidic Extracellular Polysaccharides From Rhizobium Trifolii Mutants Affected In Trifolium Repens Root Hair Curling, Brian Andrew Hanley

Masters Theses

The compositions and sugar linkages of the acidic extracellular polysaccharide (EPS) from several Rhizobium trifolii mutants were compared. These mutants were derived from a wild type Rhizobium trifolii, ANU843, and are unable to cause normal root hair curling, Hac-, or nodulate clover, Nod-. Three of the mutants contain a Tn5 insertion in a 5.3 kb BglII fragment of the symbiotic plasmid (pSym). A fourth mutant is missing the pSym, which includes much of the 5.3 kb BglII fragment, both of these deletions result in Hac-Nod- phenotypes. When the 5.3 kb BglII fragment is cloned (pRT150) and reinserted into the …