Life Sciences Commons^™

The Sos Response In Escherichia Coli K12: An Exploration Of Mutations In Lexa And Reca Using Fluorescence Microscopy, Steven Van Alstine

Doctoral Dissertations

Faithful replication of the genome is paramount for maintaining the fitness of an organism. Therefore, life has evolved inducible mechanisms to be able to repair damaged DNA and maintain evolutionary fitness. The SOS response is a highly conserved DNA damage inducible response that is tightly regulated. Multiple factors contribute to the ability of the cell to perform proper DNA repair and induction of the SOS response including the amount of RecA, mutations in RecA that affect competition for DNA, and other proteins that interact with the RecA filament. The complex relationship between RecA and LexA is the subject of this …

Screening For Binding Partners And Protein-Protein Interactions Of A Fungal Transcription Factor- Xdr1, Nishadi Punsara Gallala Gamage

Masters Theses

Clarireedia spp. (formerly Sclerotinia homoeocarpaF.T. Bennett) is the causal agent dollar spot, the most economically important turfgrass disease impacting golf courses in North America. The most effective strategy for dollar spot control is repeated application of multiple classes of fungicides. However, reliance on chemical application has led to resistance to four classes of fungicides as well as multidrug resistance (MDR). Fungi are known to detoxify xenobiotics, like fungicides, through transcriptional regulation of three detoxification phases: modification, conjugation and secretion. Little is known, however, of the protein-protein interactions that facilitate these pathways. Following next-generation RNA sequencing of Clarireedia spp., a …

The Influence Of Climate Change On The Ecology Of The Arctic Ground Squirrel In Denali National Park, Ak., Nigel A. Golden

Doctoral Dissertations

This dissertation research focuses on the ecology of the Arctic ground squirrel (Urocitellus parryii) in Denali National Park and Preserve, AK. The Arctic ground squirrels is a species of interest for monitoring efforts under the National Park Services’ Vital Signs Monitoring Program under the Vital Signs Monitoring program. The focus of this program is to monitor what is considered to be the most significant indicators of ecological conditions of the specific park resources that are of the greatest concern. The CAKN designated the Arctic ground squirrel (Urocitellus parryii) as one indicator species of park ecosystems. Despite being easy to observe …

Mapping Selected Polyphenols Metabolism By Gut Bacteria And Their Genes, Ermin Zhao

Doctoral Dissertations

The human gut microbiome is a huge enzyme repository for dietary polyphenols metabolism, especially considering most of the polyphenols cannot be digested in the host and their biological functions are limited. Poor bioaccessibility based on traditional pharmaceutical ADME (absorption, distribution, metabolism, and excretion) assessment is the main problem facing the widely medical application of most polyphenols. Gut bacteria have the potential to mediate a wide range of biotransformation reactions of polyphenols, which leads to the production of many bioactive metabolites. In the past decades, mounting evidence in traditional ADME study have demonstrated gut bacteria play an irreplaceable role in dietary …

Discovery Of Mirnas And Development Of Heat-Responsive Mirna-Ssr Markers For Characterization Of Wheat Germplasm For Terminal Heat Tolerance Breeding, Pooja Sihag, Vijeta Sagwal, Anuj Kumar, Priyanka Balyan, Reyazul Rouf Mir, Om Parkash Dhankher, Upendra Kumar

Stockbridge Faculty Publication Series

A large proportion of the Asian population fulfills their energy requirements from wheat (Triticum aestivum L.). Wheat quality and yield are critically affected by the terminal heat stress across the globe. It affects approximately 40% of the wheat-cultivating regions of the world. Therefore, there is a critical need to develop improved terminal heat-tolerant wheat varieties. Marker-assisted breeding with genic simple sequence repeats (SSR) markers have been used for developing terminal heat-tolerant wheat varieties; however, only few studies involved the use of microRNA (miRNA)-based SSR markers (miRNASSRs) in wheat, which were found as key players in various abiotic stresses. In the …

Species And Population Specific Gene Expression In Blood Transcriptomes Of Marine Turtles, Shreya M. Banerjee, Jamie Adkins Stoll, Camryn D. Allen, Jennifer M. Lynch, Heather S. Harris, Lauren Kenyon, Richard E. Connon, Eleanor J. Sterling, Eugenia Naro-Maciel, Kathryn Mcfadden

Environmental Conservation Faculty Publication Series

Background: Transcriptomic data has demonstrated utility to advance the study of physiological diversity and organisms' responses to environmental stressors. However, a lack of genomic resources and challenges associated with collecting high-quality RNA can limit its application for many wild populations. Minimally invasive blood sampling combined with de novo transcriptomic approaches has great potential to alleviate these barriers. Here, we advance these goals for marine turtles by generating high quality de novo blood transcriptome assemblies to characterize functional diversity and compare global transcriptional profiles between tissues, species, and foraging aggregations.ResultsWe generated high quality blood transcriptome assemblies for hawksbill (Eretmochelys imbricata), loggerhead …

Navigating The U.S. Health Insurance Landscape For Children With Rare Diseases: A Qualitative Study Of Parents' Experiences, Tai L. S. Pasquini, Sarah L. Goff, Jennifer M. Whitehill

Health Promotion and Policy Faculty Publication Series

Parents of children with rare diseases often face uncertainty about diagnosis, treatment, and costs associated with healthcare for their child. Health insurance status impacts each of these areas, but no U.S. study has explored parents' perceptions of the health insurance impacts on their child's care. This study aimed to qualitatively explore how these parents navigate the complex health insurance system for their children and their experiences in doing so. Methods Semi-structured interviews were conducted with parents of children with metachromatic leukodystrophy (MLD) and spinal muscular atrophy (SMA), chosen for specific disease characteristics and orphan drug status. Participants were recruited via …

Population Viability And Connectivity Of The Federally Threatened Eastern Indigo Snake In Central Peninsular Florida, Javan Bauder

Doctoral Dissertations

Understanding the factors influencing the likelihood of persistence of real-world populations requires both an accurate understanding of the traits and behaviors of individuals within those populations (e.g., movement, habitat selection, survival, fecundity, dispersal) but also an understanding of how those traits and behaviors are influenced by landscape features. The federally threatened eastern indigo snake (EIS, Drymarchon couperi) has declined throughout its range primarily due to anthropogenically-induced habitat loss and fragmentation making spatially-explicit assessments of population viability and connectivity essential for understanding its current status and directing future conservation efforts. The primary goal of my dissertation was to understand how …

Protein Degradation Regulates Phospholipid Biosynthetic Gene Expression In Saccharomyces Cerevisiae, Bryan Salas-Santiago

Doctoral Dissertations

Transcriptional regulation of most phospholipid biosynthetic genes in Saccharomyces cerevisiae is coordinated by inositol and choline. Inositol affects phosphatidic acid (PA) intracellular levels. Opi1p interacts physically with PA and is the main repressor of the phospholipid biosynthetic genes. It is localized in the endoplasmic reticulum (ER) bound to the ER membrane protein Scs2p. When PA levels drop, Opi1p is translocated into the nucleus repressing most phospholipid biosynthetic genes. The OPI1 locus was identified in a screen looking for overproduction and excretion of inositol (Opi^-). Opi^- mutants are generally associated with a defect in …

An Integrative Approach To Understanding Morphological Novelties: Anatomy, Development, Genetics, And Evolution Of An Extreme Craniofacial Trait In East African Cichlids, Moira R. Conith

Doctoral Dissertations

Phenotypic novelties are an important but poorly understood category of morphological diversity that are often associated with elevated rates of diversification and/or ecological success. The aim of this dissertation is to explore a phenotypic novelty at many levels to contribute to our understanding of how these unique traits can arise (e.g., genetically, developmentally, and evolutionarily) as well as their ecological consequences (e.g., trait function). The extreme snout of the Lake Malawi cichlid fish Labeotropheus is used as a case study. The first chapter establishes the Labeotropheus snout as a model of phenotypic novelty by characterizing the gross morphology, genetic architecture, …

Variation And Evolution Of Fruit Ripening Traits In Tomato Species, Ian M. Gillis

Doctoral Dissertations

As angiosperm seeds mature within their ovaries, ovary tissue tends to grow and transform itself into fruit, which aids the success of the seeds. Fruits that are fleshy provide numerous ways to aid in the protection and the dispersal of seeds. First, they keep seeds hidden, encased in hard walls, surrounded by poisons and unpalatable compounds, and second, they undergo developmental changes that facilitate seeds’ release. Tomatoes, a model fleshy fruit, have all these protective traits, and over the course of ripening they become the familiar fruit that is a staple crop around the world. The wild relatives of cultivated …

Clpxp-Regulated Proteins Suppress Requirement For Reca In Dam Mutants Of Escherichia Coli K-12, Amie Savakis

Masters Theses

Double strand breaks (DSB) are a common source of DNA damage in both prokaryotes and eukaryotes. If they are not repaired or are repaired incorrectly, they can lead to cell death (bacteria) or cancer (humans). In Escherichia coli, repair of DSB are typically accomplished via homologous recombination and mediated by RecA. This repair pathway, among others, is associated with activation of the SOS response. DNA adenine methyltransferase (dam) mutants have an increased number of DSB and, therefore, are notorious for being RecA-dependent for viability. Here, we show that the synthetic lethality of Δdam/ΔrecA is suppressed when clpP is removed, suggesting …

Characterization Of She1 Spindle Role Using Ceullular, Biochemical, And Biophysical Methods, Yili Zhu

Doctoral Dissertations

During development, metaphase spindles undergo large movement and/or rotation to determine the cell division axis. While it has been shown that spindle translocation is achieved by astral microtubules pulling and/or pushing the cortex, how metaphase spindle stability is maintained during translocation remains not fully understood. In budding yeast, our lab has previously proposed a model for spindle orientation wherein the mitotic spindle protein She1 promotes spindle translocation across the bud neck by polarizing cortical dynein pulling activity on the astral microtubules. Intriguingly, She1 exhibits dominant spindle localization throughout the cell cycle. However, whether She1 has any additional role on the …

Impacts Of Genome And Nuclear Architecture On Molecular Evolution In Eukaryotes, Xyrus Maurer-Alcalá

Doctoral Dissertations

The traditional view of genomes suggests that they are static entities changing slowly in sequence and structure through time (e.g. evolving over geological time-scales). This outdated view has been challenged as our understanding of the dynamic nature of genomes has increased. Changes in DNA content (i.e. polyploidy) are common to specific life-cycle stages in a variety of eukaryotes, as are changes in genome content itself. These dramatic genomic changes include chromosomal deletions (i.e. paternal chromosome deletion in insects; Goday and Esteban 2001; Ross, et al. 2010), developmentally regulated genome rearrangements (e.g. the V(D)J system in adaptive immunity in mammals; Schatz …

All Roads Lead To Weediness: Stories About Weedy Rice Origins, Weedy Genes And Weed Competitiveness, Zhongyun Huang

Doctoral Dissertations

Weedy rice (Oryza spp.), a weedy relative of cultivated rice (O.sativa), infests and persists in cultivated rice fields worldwide. Many weedy rice populations have evolved similar adaptive traits, considered part of the ‘agricultural weed syndrome’, making this an ideal model to study the genetic basis of parallel evolution. Using population genetics analyses of South Asian and US weedy rice, my research reveals multiple independent evolution events giving rise to weed groups in the two geographic areas. Weeds in South Asia have highly heterogenous genetic backgrounds, with contributions from both cultivated varieties (aus and indica) …

Uncovering Tasselsheath3. A Genomic And Phenotypic Analysis Of A Maize Floral Mutant., Thompson Zhang

Masters Theses

In the modern era, maize has become the most successful crop grown in the United States. According to the USDA over 90 million acres of land are planted to corn and 96.2% of the U.S feed grain production is made up of the cereal. Part of the success of maize is due to its floral architecture, and its pollination technique in which the flower opens, exposing stamens containing pollen into the air. A unique organ called the lodicule functions as a release mechanism, forcing the flower to open. Lodicules from grasses and eudicot petals are homologous, yet there is little …

Epigallocatechin-3-Gallate Reduces Fat Accumulation In Caenorhabditis Elegans, Jinning Liu

Masters Theses

Epigallocatechin gallate (EGCG), also known as epigallocatechin-3-gallate, is a polyphenol that is most abundant in tea. It has been shown from many studies that consumption of EGCG can contribute to weight loss, however, the underlying mechanism is not fully understood. To determine how EGCG acts to reduce fat, an organism model Caenorhabditis elegans (C. elegans) is introduced, which is a useful animal system in exploring crucial biological mechanisms that are readily applicable to humans. In this study, different strains were raised for two days on a diet with or without 100µM and 200µM EGCG treatment: N2 (i.e., wild …

The Key Question In Symbiotic Nitrogen Fixation: How Does Host Maintain A Bacterial Symbiont?, Onur Oztas

Doctoral Dissertations

The fact that plants cannot use nitrogen in the gaseous form makes them dependent on the levels of usable nitrogen forms in the soil. Legumes overcome nitrogen limitation by entering a symbiotic association with rhizobia, soil bacteria that convert atmospheric nitrogen into usable ammonia. In root nodules, bacteria are internalized by host plant cells inside an intracellular compartment called the symbiosome where they morphologically differentiate into nitrogen-fixing forms by symbiosome-secreted host proteins. In this project, I explained the host proteins required to maintain bacterial symbionts and described their delivery to the symbiosome. I showed that the SYNTAXIN 132 (SYP132) gene …

A Genetic Analysis Of Cichlid Scale Morphology, Kenta C. Kawasaki

Masters Theses

Epidermal appendages are found on every vertebrate this world has to offer. In fish, these are commonly represented by scales. While we have a solid grasp of how scales develop, little is known about the underlying genetic mechanisms behind these phenotypic changes. Using two species of African cichlids (Labeotropheus fuelleborni and Tropheops “red cheek”) with varying scale phenotypes, we sought to examine their F2 hybrid offspring and statistically link the responsible genetic elements to their respective parental phenotypes through Quantitative Loci Trait (QTL) analysis.

Scales were removed from six different locations across the midline of each individual. Then, numerous …

Population Genetic Analysis Of Atlantic Horseshoe Crabs (Limulus Polyphemus) In Coastal Massachusetts., Katherine T. Johnson

Masters Theses

Atlantic horseshoe crabs (Limulus polyphemus) have endured decades of intense harvest pressure. Genetics studies have shown evidence of distinct sub-groups spanning the coast, although few fine-scale studies have been done to delineate these groups on a local level. Massachusetts lies directly between two of these sub-groups. With documented differences in prosomal widths of horseshoe crabs from either side of Cape Cod, it is possible that Cape Cod is a barrier to gene flow and that there are two distinct genetic groups within Massachusetts. Regulations currently consider all horseshoe crabs to be of one stock. I examined 6 microsatellite …

Evolvability Of The Skull: A Study Of Genetic Basis And Integration In The Teleost Craniofacial Skeleton, Yinan Hu

Doctoral Dissertations

As the field of evolutionary biology pivots away from a gene-centric view of how adaptive evolution proceeds, renewed emphasis is placed on the origin of phenotypic variation. Understanding the developmental processes that underlie the production of novel traits, and how they might influence evolvability, is considered a primary goal in the on-going “extended evolutionary synthesis”. The following dissertation explores these questions in the context of adaptive radiations in fish, with a focus on morphological variation in the craniofacial skeleton. Specifically, the first chapter investigates the genetic and developmental basis of shape (co-)variation in the feeding apparatus of African cichlid fishes, …

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 …

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 …

Evolution Of Hybrid Incompatibilities In Gene Regulatory Networks, Alexander Y. Tulchinsky

Open Access Dissertations

Under the Dobzhansky-Muller model, postzygotic isolation results from incompatibility between interacting genes. Evidence points to regulatory networks as a rich source of incompatibilities that impact hybrid fitness. Pleiotropy is a natural feature of regulatory networks because regulatory elements generally have multiple targets. Both pleiotropy and hybrid incompatibility arise due to genetic interactions; therefore we can expect an intimate association between them. In the following chapters, I investigate the relationship between pleiotropy and hybrid incompatibility in the context of regulatory networks.

In chapter one, I extend a general network-based study of hybrid incompatibility by incorporating a sequence-based thermodynamic model of transcriptional …

Gene Number Determination And Genetic Polymorphism Of The Gamma Delta T Cell Co-Receptor Wc1 Genes, Chuang Chen, Carolyn Ta Herzig, Leeson J. Alexander, John W. Keele, Tara G. Mcdaneld, Janice C. Telfer, Cynthia L. Brown

Janice C. Telfer

Background: WC1 co-receptors belong to the scavenger receptor cysteine-rich (SRCR) superfamily and are encoded by a multi-gene family. Expression of particular WC1 genes defines functional subpopulations of WC1+ γδ T cells. We have previously identified partial or complete genomic sequences for thirteen different WC1 genes through annotation of the bovine genome Btau_3.1 build. We also identified two WC1 cDNA sequences from other cattle that did not correspond to sequences in the Btau_3.1 build. Their absence in the Btau_3.1 build may have reflected gaps in the genome assembly or polymorphisms among animals. Since the response of γδ T cells to bacterial …

Identification Of Pmt, Tr1, And H6h Gene Polymorphism And Tropane Alkaloid Chemotypes In Hyoscyamus Niger L. (Black Henbane), Lawrence Kramer

Masters Theses 1911 - February 2014

No abstract provided.

Land Use Laws And Policies Model Biogenetics Bylaw, Umass Amherst Center Economic Development

Center for Economic Development Technical Reports

This report explains the bylaw that dictates the use of RDNA in the town of Grafton, Massachusetts.