Genetics and Genomics Commons^™

Evolutionary Patterns In Deep-Sea Mollusks, Elizabeth Ellen Boyle

Graduate Doctoral Dissertations

The evolution of deep-sea fauna is poorly known. We know little about the scales of population differentiation, radiation and colonization of the deep sea. This dissertation explores evolution in the deep sea at the level of population differentiation on ocean-wide scales in a gastropod species Benthonella tenella and phylogeny of a deep-sea subfamily of protobranch bivalves, Ledellinae. While working on the phylogeny of the Ledellinae the presence of mitochondrial heteroplasmy was discovered.

Genetic variation was quantified within and among populations of deep-sea gastropod Benthonella tenella to identify the extent of population differentiation and potential mechanisms that might isolate gene pools. …

Removing Technical Variability In Rna-Seq Data Using Conditional Quantile Normalization, Kasper D. Hansen, Rafael A. Irizarry, Zhijin Wu

Johns Hopkins University, Dept. of Biostatistics Working Papers

The ability to measure gene expression on a genome-wide scale is one of the most promising accomplishments in molecular biology. Microarrays, the technology that first permitted this, were riddled with problems due to unwanted sources of variability. Many of these problems are now mitigated, after a decade’s worth of statistical methodology development. The recently developed RNA sequencing (RNA-seq) technology has generated much excitement in part due to claims of reduced variability in comparison to microarrays. However, we show RNA-seq data demonstrates unwanted and obscuring variability similar to what was first observed in microarrays. In particular, we find GC-content has a …

Recombination Phenotypes Of The Nci-60 Collection Of Human Cancer Cells, Dawn M. Stults, Michael W. Killen, Brent J. Shelton, Andrew J. Pierce

Microbiology, Immunology, and Molecular Genetics Faculty Publications

BACKGROUND: The NCI-60 is a collection of tumor cell lines derived from a variety of human adult cancer tissue types and is commonly used for genetic analysis and screening of potential chemotherapeutic agents. We wanted to understand the contributions of specific mechanisms of genomic instability to the etiology of cancers represented by the NCI-60.

RESULTS: We screened the NCI-60 for dysregulated homologous recombination by using the gene cluster instability (GCI) assay we pioneered, and for defects in base excision repair by sensitivity to 5-hydroxymethyl-2'-deoxyuridine (hmdUrd). We identified subsets of the NCI-60 lines that either displayed the characteristic molecular signature of …

Expression Analysis Of The Imprinted Gene Transketolase-Like 1 In Mouse And Human, Amy F. Friss

Master's Theses

Genomic imprinting is an epigenetic phenomenon resulting in differential gene expression based on parental origin. Recently, transketolase-like 1 (TKTL1) has been identified as an X-linked imprinted gene. TKTL1 functions in the nonoxidative branch of the pentose phosphate pathway (PPP), which maintains glutathione in a reduced state through the generation of NADPH. Previous studies on transaldolase, the other critical enzyme in the nonoxidative branch of the PPP, suggest that TKTL1 may affect the cell’s ability to reduce glutathione. This study provides evidence that TKTL1 overexpression inhibits glutathione reduction. Intriguingly, aberrant glutathione levels are associated with autism. Additionally, studies involving …

Phylogeography Of The Polychaete Phragmatopoma Californica (Family Sabellarididae) In The Southern California Bight (Scb), Praneeth Kamana

Student Theses and Dissertations

This study measured the genetic differentiation among populations of Phragmatopoma californica and compared this genetic structuring with another polychaete species (Neries grubei) having a different life history. A total of 234 samples from 13 populations of the sabelleriid polychaete Phragmatapoma californica (Fewkes) from six mainland sites along the southern California coast and two Channel Islands were studied by way of inter simple sequence repeats (ISSRs) to determine genetic diversity, divergence and to estimate gene flow among populations. The results were compared to another polycheate species, Neries grubei which has a shorter larval development than P. californica, reducing potential dispersal and …

Drosophila Model For Fetal Alcohol Syndrome Disorders: Role For The Insulin Pathway, Rachael L. French, K D. Mcclure, U Heberlein

Faculty Publications, Biological Sciences

Prenatal exposure to ethanol in humans results in a wide range of developmental abnormalities, including growth deficiency, developmental delay, reduced brain size, permanent neurobehavioral abnormalities and fetal death. Here we describe the use of Drosophila melanogaster as a model for exploring the effects of ethanol exposure on development and behavior. We show that developmental ethanol exposure causes reduced viability, developmental delay and reduced adult body size. We find that flies reared on ethanol-containing food have smaller brains and imaginal discs, which is due to reduced cell division rather than increased apoptosis. Additionally, we show that, as in mammals, flies reared …

Expression And Purification Of Recombinant Hemoglobin In Escherichia Coli, Chandrasekhar Natarajan, Xiaoben Jiang, Angela Fago, Roy E. Weber, Hideaki Moriyama, Jay F. Storz

Jay F. Storz Publications

Background: Recombinant DNA technologies have played a pivotal role in the elucidation of structure-function relationships in hemoglobin (Hb) and other globin proteins. Here we describe the development of a plasmid expression system to synthesize recombinant Hbs in Escherichia coli, and we describe a protocol for expressing Hbs with low intrinsic solubilities. Since the alpha- and beta-chain Hbs of different species span a broad range of solubilities, experimental protocols that have been optimized for expressing recombinant human HbA may often prove unsuitable for the recombinant expression of wildtype and mutant Hbs of other species.

Methodology/Principal Findings: As a test case …

Construction Of Papaya Male And Female Bac Libraries And Application In Physical Mapping Of The Sex Chromosomes, Andrea R. Gschwend, Qingyi Yu, Paul Moore, Christopher Saski, Cuixia Chen, Jianping Wang, Jong-Kuk Na, Ray Ming

Publications

Papaya is a major fruit crop in the tropics and has recently evolved sex chromosomes. Towards sequencing the papaya sex chromosomes, two bacterial artificial chromosome (BAC) libraries were constructed from papaya male and female genomic DNA. The female BAC library was constructed using restriction enzyme BstY I and consists of 36,864 clones with an average insert size of 104 kb, providing 10.3x genome equivalents. The male BAC library was constructed using restriction enzyme EcoR I and consists of 55,296 clones with an average insert size of 101 kb, providing 15.0x genome equivalents. The male BAC library was used in constructing …

The Role Of Historical And Contemporary Processes On Phylogeographic Structure And Genetic Diversity In The Northern Cardinal, Cardinalis Cardinalis, Brian T. Smith, Patricia Escalante, Blanca E. Hernandez-Banos, Adolfo G. Navarro-Siguenza, Sievert Rohwer, John Klicka

Ornithology Program (HRC)

Background

Earth history events such as climate change are believed to have played a major role in shaping patterns of genetic structure and diversity in species. However, there is a lag between the time of historical events and the collection of present-day samples that are used to infer contemporary population structure. During this lag phase contemporary processes such as dispersal or non-random mating can erase or reinforce population differences generated by historical events. In this study we evaluate the role of both historical and contemporary processes on the phylogeography of a widespread North American songbird, the Northern Cardinal, Cardinalis cardinalis …

The Basis And Implications Of Genetic Counseling In Patient Testing Choice, Yanina Tovpeko

Renée Crown University Honors Thesis Projects - All

Perinatal genetic counseling is a health service provided to patients who carry risk factors for genetic abnormalities for their offspring. There are several non-invasive and invasive tests offered to patients in order to provide genetic information about possible disorders. The non-invasive screens calculate a percentage of risk relative to average population risk. If the non-invasive screens predict an elevated risk, a more invasive test can be offered to obtain definitive results about possible disorders in the pregnancy. The most commonly used invasive test is called amniocentesis and it carries a risk for pregnancy complications. After receiving abnormal results on the …

Analysis Of Long-Term Selection (28 Generations) For Reproduction, Growth, And Carcass Traits In Swine, Wan-Ling Hsu

Department of Animal Science: Dissertations, Theses, and Student Research

The objectives were 1) to estimate responses from 28 generations of selection for increased pigs born alive (BA) with additional selection for increased 180 d weight (WT) and longissimus muscle area (LMA) and decreased backfat (BF) in the last eight generations, 2) to estimate genetic parameters for an array of traits, and 3) to investigate whether a plateau in response for BA has occurred. All lines were derived from the same Large White/Landrace composite population. Index selection for ovulation rate and embryo survival (G0-11) was initiated in L2 in 1981; L1 was selected randomly control line for L2 (G0-23). L2 …

Population Genetics Of The Western Bean Cutworm (Striacosta Albicosta Smith) Across The United States, Erica Lindroth

Department of Entomology: Dissertations, Theses, and Student Research

The western bean cutworm, Striacosta albicosta (Smith), is a secondary pest of maize (Zea mays L.) and dry beans (Phaseolus vulgaris L.) in the western United States. Recently, this insect has undergone a major territory expansion into the eastern US and has become a pest throughout much of the Corn Belt. This study was instigated to examine the population genetics of this pest to facilitate control and resistance management, as well as to shed light on the current habitat expansion. To this end, western bean cutworm individuals were collected from 28 different locations across the traditional and expanded …

Design Of A Factorial Experiment With Randomization Restrictions To Assess Medical Device Performance On Vascular Tissue, Wiebke Diestelkamp, Carissa M. Krane, Margaret Pinnell

Biology Faculty Publications

Background: Energy-based surgical scalpels are designed to efficiently transect and seal blood vessels using thermal energy to promote protein denaturation and coagulation. Assessment and design improvement of ultrasonic scalpel performance relies on both in vivo and ex vivo testing. The objective of this work was to design and implement a robust, experimental test matrix with randomization restrictions and predictive statistical power, which allowed for identification of those experimental variables that may affect the quality of the seal obtained ex vivo.

Methods: The design of the experiment included three factors: temperature (two levels); the type of solution used to perfuse the …

Endo-Porter–Mediated Delivery Of Phosphorodiamidate Morpholino Oligos (Pmos) In Erythrocyte Suspension Cultures From Cope's Gray Treefrog Hyla Chrysoscelis, Venkateshwar Mutyam, Matthew V. Puccetti, James Frisbie, David L. Goldstein, Carissa M. Krane

Biology Faculty Publications

Cope's gray treefrog, Hyla chrysoscelis, is a freeze-tolerant anuran that accumulates cryoprotective glycerol during cold acclimation. H. chrysoscelis erythrocytes express the aquaglyceroporin HC-3, which facilitates transmembrane glycerol and water movement. Aquaglyceroporins have no pharmacological inhibitors, and no genetic knockout tools currently exist for H. chrysoscelis. A phosphorodiamidate morpholino oligo (PMO)–mediated expression knockdown approach was therefore pursued to provide a model for testing the role of HC-3. We describe a novel procedure optimized for specific, efficient knockdown of HC-3 expression in amphibian erythrocyte suspensions cultured at nonmammalian physiological temperatures using Endo-Porter. Our protocol includes three critical components: pre-incubation at 37°C, …

Microarray Data Mining And Gene Regulatory Network Analysis, Ying Li

Dissertations

The novel molecular biological technology, microarray, makes it feasible to obtain quantitative measurements of expression of thousands of genes present in a biological sample simultaneously. Genome-wide expression data generated from this technology are promising to uncover the implicit, previously unknown biological knowledge. In this study, several problems about microarray data mining techniques were investigated, including feature(gene) selection, classifier genes identification, generation of reference genetic interaction network for non-model organisms and gene regulatory network reconstruction using time-series gene expression data. The limitations of most of the existing computational models employed to infer gene regulatory network lie in that they either suffer …

Phylogenetic Diversification Of The Globin Gene Superfamily In Chordates, Jay F. Storz, Juan C. Opazo, Federico G. Hoffmann

Jay F. Storz Publications

Phylogenetic reconstructions provide a means of inferring the branching relationships among members of multigene families that have diversified via successive rounds of gene duplication and divergence. Such reconstructions can illuminate the pathways by which particular expression patterns and protein functions evolved. For example, phylogenetic analyses can reveal cases in which similar expression patterns or functional properties evolved independently in different lineages, either through convergence, parallelism, or evolutionary reversals. The purpose of this article is to provide a robust phylogenetic framework for interpreting experimental data and for generating hypotheses about the functional evolution of globin proteins in chordate animals. To do …

Cocaine Dependence: The Role Of Serotonin Genes In, Lorena Maili

Dissertations and Theses (Open Access)

Animal studies have shown that behavioral responses to cocaine-related cues are altered by serotonergic medications. The effects of pharmacological agents on serotonin receptors 2a (5-HT2A) and 2c (5-HT2C), have yielded results suggesting that selective 5-HT2A antagonists and 5-HT2C agonists promote the disruption of cocaine-associated memories. One measure of cocaine related cues in humans is attentional bias, in which cocaine dependent individuals show greater response latency for cocaine related words than neutral words. Data from our laboratory shows that cocaine dependent subjects have altered attentional bias compared to controls. The purpose of this thesis was to investigate the role of the …

The Role And Mechanism Of The Homeobox Gene Dlx4 In Transforming Growth Factor-B Resistance In Cancer, Bon Q. Trinh

Dissertations and Theses (Open Access)

Transforming growth factor-b (TGF-b) is a cytokine that plays essential roles in regulating embryonic development and tissue homeostasis. In normal cells, TGF-b exerts an anti-proliferative effect. TGF-b inhibits cell growth by controlling a cytostatic program that includes activation of the cyclin-dependent kinase inhibitors p15^Ink4B and p21^WAF1/Cip1 and repression of c-myc. In contrast to normal cells, many tumors are resistant to the anti-proliferative effect of TGF-b. In several types of tumors, particularly those of gastrointestinal origin, resistance to the anti-proliferative effect of TGF-b has been attributed to TGF-b receptor or Smad mutations. However, these mutations are absent from many …

Cell Polarity Regulates Organ Growth Through The Hippo Pathway, Chiao-Lin Chen

Dissertations and Theses (Open Access)

Defects in apical-basal cell polarity and abnormal expression of cell polarity determinants are linked to human cancer. Loss of polarity is highly correlated with malignancy. In Drosophila, perturbation of apical-basal polarity, including overexpressing the apical determinant Crumbs, can lead to uncontrolled tissue growth. Cells mutant for the basolateral determinant scribble overproliferate and can form neoplastic tumors. Interestingly, scribble mutant clones that arise in wild-type tissues are eliminated and therefore do not manifest their tumorigenic potential. However, the mechanisms by which cell polarity coordinates with growth control pathways in developing organs to achieve appropriate organ size remain obscure.

To investigate …

Gene Discovery In Nonsyndromic Cleft Lip With Or Without Cleft Palate, Brett T. Chiquet

Dissertations and Theses (Open Access)

Nonsyndromic cleft lip with or without cleft palate (NSCLP), a common, complex orofacial birth defect that affects approximately 4,000 newborns each year in the United States, is caused by both genetic and environmental factors. Orofacial clefts affect the mouth and nose, causing severe deformity of the face, which require medical, dental and speech therapies. Despite having substantial genetic liability, less than 25% of the genetic contribute to NSCLP has been identified. The studies described in this thesis were performed to identify genes that contribute to NSCLP and to demonstrate the role of these genes in normal craniofacial development. Using genome …