Microbiology Commons^™

Acetobacter Fabarum Genes Influencing Drosophila Melanogaster Phenotypes, Kylie Makay White

Theses and Dissertations

Research in our lab has predicted hundreds of bacterial genes that influence nine different traits in the fruit fly, Drosophila melanogaster. As a practical alternative to creating site-directed mutants for each of the predicted genes, we created an arrayed transposon insertion library using a strain of Acetobacter fabarum DsW_054 isolated from fruit flies. Creation of the Acetobacter fabarum DsW_054 gene knock-out library was done through random transposon insertion, combinatorial mapping and Illumina sequencing. Successful mapping of transposon insertion was achieved for 6418 mutants with hits within 63% of annotated genes within Acetobacter fabarum DsW_054. Insertion sites were verified in …

Influence Of Epstein-Barr Virus On Systemic Lupus Erythematosus Disease Development And The Role Of Depression On Disease Progression, Caleb Cornaby

Theses and Dissertations

Systemic Lupus Erythematosus (SLE) is an autoimmune disease affecting 20 to 250 individuals per 100,000 worldwide. Symptomology includes dermatological manifestations such as discoid lesions, acute cutaneous rashes, and oral and nasal ulcers, along with musculoskeletal, pulmonary, and renal complications. Abnormal T and B lymphocyte function and apoptosis, immune complex clearance, complement function, and nucleosome processing are typical of disease pathophysiology. SLE is the result of both environmental and genetic factors, which together create the conditions leading to disease onset and progression. Of these environmental factors, Epstein-Barr virus (EBV) infection is known to cause the genesis of cross-reactive antibodies in SLE …

Phosphate Signaling Through Alternate Conformations Of The Pstscab Phosphate Transporter, Ramesh Krishna Vuppada

Theses and Dissertations

Phosphate is an essential compound for life. Escherichia coli employs a signal transduction pathway that controls the expression of genes that are required for the high-affinity acquisition of phosphate and the utilization of alternate sources of phosphorous. These genes are only expressed when environmental phosphate is limiting. The seven genes for this signaling pathway encode the two-component regulatory proteins PhoB and PhoR, as well as the high-affinity phosphate transporter PstSCAB and an auxiliary protein called PhoU. As the sensor kinase PhoR has no periplasmic sensory domain, the mechanism by which these cells sense environmental phosphate is not known. This paper …

Autoimmunity-Promoting B Cell Effects Of The Risk Factor Irf5, Brian Poole

Journal of Undergraduate Research

The academic objectives of this proposal were met. Using the funding provided by the MEG, we generated data that were used to publish two peer-reviewed journal articles:

Final Report For The 2015 Meg Entitled: Development Of Real-Time Polymerase Chain Reaction (Q-Pcr) Assays For The Detection And Identification Of Human Tick-Borne Pathogens, Richard Robison

Journal of Undergraduate Research

1. Develop singleplex q-PCR assays to identify the tick-borne pathogens Borrelia burgdorferi, Borrelia hermsii, Bartonella henselae, and Babesia microti. Primer generation software will be used to design primer and probe sequences that will theoretically perform optimally in PCR reactions. All sequences will be subjected to BLAST searches to confirm their specificity for a particular organism. Once primer sequences have been obtained, they will be synthesized by Integrated DNA Technologies, Inc. (IDT). These primers will be evaluated in q-PCR reactions using target DNA from multiple isolates and SyberGreen to detect nucleic acid amplification. Parameters for each reaction will be optimized using these …

The Chemotherapeutic Potential Of Resveratrol-Based Treatments, Connor Peck, Kim O'Neill

Journal of Undergraduate Research

Cancer is one of the most prevalent and deadly diseases in the world. In 2012 alone, there were approximately 14 million new cases of cancer and over 8 million cancer-related deaths worldwide. Despite extensive research in past decades, the burden of cancer continues to increase.

One of the most common treatments for cancer is chemotherapy, the use of various chemical substances that exhibit anti-cancer activity. Unfortunately, chemotherapy can be quite toxic and often produces inadequate results. There remains a great need for improved chemotherapeutic treatment strategies in order to improve outcomes for cancer patients.

The purpose of my project was …

Effect Of High Expression Of Irf5 On B Cell Activation After Antigen Stimulation, Kalarelyn Eberting, Brian Poole

Journal of Undergraduate Research

While autoimmune diseases are very prevalent in the United States, little is understood about the cause of autoimmunity, making it difficult to take preventative measures and develop treatments. Previous studies have determined that a common genetic risk factor across many autoimmune diseases leads to B cell overexpression of the transcription factor IRF5 (1,2,4). Researchers have also discovered that high IRF5 levels are strongly associated with autoantibody production, suggesting that IRF5 may play a role in actually causing autoimmunity. At this point, however, it is unclear how IRF5 promotes the production of autoantibodies.

Development Of A Model For Adult T Cell Leukemia In Htlv-1 Infected Humanized Mice, Matthew Smith, Dr. Brad Berges

Journal of Undergraduate Research

Millions of people in the world are infected with human T-lymphotropic virus type-1 (HTLV-1). As the name states, this virus attacks the T cells of the infected organism. Though many of these individuals will remain asymptomatic throughout life, a small percentage will develop adult T cell leukemia/lymphoma (ATLL), a rare and understudied form of cancer. The latency period, or amount of time between infection and detection of cancer, is generally 20-40 years before infected patients begin developing cancer.

Phenome-Wide Association Study Of Y Chromosome Genetic Markers, Matthew Durrant, Mary Davis

Journal of Undergraduate Research

The purpose of this project was to analyze genetic variants on the Y chromosome for significant association with various diseases. In Dr. Mary Davis’ lab we gained access to a unique data set that allows us to analyze many single-nucleotide polymorphisms (SNPs) on the Y chromosome. This was a hypothesis-generating study that has the potential to revolutionize the currently accepted hypothesis that genetic variants on the Y chromosome do not play a significant role in common disease.

Cellular Mechanism Of Thymidine Kinase 1 Secretion, Wei Meng, Kim O'Neill

Journal of Undergraduate Research

Cancer remains the second most common cause of death in the US, accounting for nearly 1 of every 4 deaths. Studies have shown that the early detection of cancer leads to better patient prognosis and a greater five-year survival rate. [1] Diagnostic and prognostic markers play a key role in classifying tumors and determining the best treatment plan for a patient. Among these biomarkers, Thymidine kinase 1 (TK1) has been studied extensively, primarily as a diagnostic biomarker for a variety of cancer types. TK1 plays a role in regulating the intracellular thymidine pools throughout the cell cycle, and thus regulates …

Expression Changes Of Dopamine Type 2 Receptor On The Surface Of Human Monocytes And Lymphocytes After Exposure To Dopamine In Vitro., Brandon Thomas Garcia, Sandra Hope, Scott Steffensen

Journal of Undergraduate Research

Dopamine (DA) is a neurotransmitter in the brain that plays a role in several functions such as reward, motivation, and dependence. The imbalance of DA in the central nervous system (CNS) can lead to several debilitating diseases like Parkinson’s Disease, Alzheimer’s, and Schizophrenia as well as drug dependence. Levels of DA in the brain are often determined by measuring the expression of Dopamine type 2 (D2) receptors by positron emission topography (PET) scans in the Nucleus Accumbens (NAc). This test which is often expensive and unavailable to many people without access to hospitals or regular healthcare.

Development Of Chimeric Antigen Receptor T Cells To Target Cancer, Justin Crandall, Dr. Scott Weber

Journal of Undergraduate Research

Cancer negatively affects the lives of millions of individuals, and remains a difficult ailment to treat. In 2014, approximately 585,720 deaths and 1,665,540 new cases of cancer were reported in the USA alone.1 Treatment of cancer is challenging due to cancer mutating to combat the body’s immune system. An example is shown in major histocompatibility complexes (MHCs) located on the surface of nearly every cell. These are patrolled by T cells to ensure that cells are properly functioning and healthy. With cancer however, these become downregulated allowing the cancer to grow and proliferate since T cells are unable to detect …

Creation Of Twelve Member Plasmid Library For Promoter Swapping To Control Chromosomal Gene Expression In E. Coli, Chris Nielson, William R. Mccleary

Journal of Undergraduate Research

Metabolic engineering is becoming a very important area of research, allowing researchers to harness metabolic pathways to either eliminate or synthesize desired compounds. Understanding metabolic pathways by altering expression of proteins involved in the pathway helps to uncover thermodynamic bottlenecks which render certain pathways inefficient or infeasible. Altering expression of genes through the use of promoter swapping is a useful research technique used to understand important metabolic and physiologic pathways. Engineered promoters contained in plasmids can be inserted into chromosomal DNA through the utilization of promoter swapping to alter the expression of a given gene in order to better understand …

The Effects Of Nucleosome Positioning And Chromatin Architecture On Transgene Expression, Colton E. Kempton

Theses and Dissertations

Eukaryotes use proteins to carefully package and compact their genomes to fit into the nuclei of their individual cells. Nucleosomes are the primary level of compaction. Nucleosomes are formed when DNA wraps around an octamer of histone proteins and a nucleosome's position can limit access to genetic regulatory elements. Therefore, nucleosomes represent a basic level of gene regulation. DNA and its associated proteins, called chromatin, is usually classified as euchromatin or heterochromatin. Euchromatin is transcriptionally active with loosely packed nucleosomes while heterochromatin is condensed with tightly packed nucleosomes and is transcriptionally silent. In order to become active, heterochromatin must first …

Thymidine Kinase 1: The Key To Predicting Cancer Recurrence, Rachel Brog, Dr. Kim L. O'Neill

Journal of Undergraduate Research

Currently it is estimated that over 230,000 women will be diagnosed with breast cancer this year in the United States alone. Although treatments are improving, an alarming number of these cases will show cancer recurrence after treatment. A method to consistently predict cancer recurrence is crucial to better treat cancer and improve patient care. Thymidine Kinase 1 (TK1) has been found in abnormally high levels in serum and tissue of cancer patients compared to healthy patients. There is question to why TK1 is present in such high quantities when normally it aids in the DNA salvage pathway during S phase …

Characterization Of Neuronal Nicotinic Acetylcholine Receptors And Their Positive Allosteric Modulators, Doris Clark Jackson

Theses and Dissertations

Neuronal nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that are necessary in memory and cognition. They are pentameric and consist of α and β subunits. They are most commonly heteromeric but, can sometimes be homomeric. nAChRs are activated by many ligands including nicotine (exogenous) and acetylcholine (endogenous).nAChRs are located on hippocampal interneurons. The interneurons, although sparse, control the synchronous firing of the pyramidal cells. However, the hippocampal interneuron structure and function is quite diverse and not fully characterized. Therefore, we sought to quantify nAChR subunit mRNA levels using real-time PCR of CA1 hippocampal interneurons.Surprisingly we found that the α3 …

Comparison Of Cytokine Expression And Bacterial Growth During Periparturient And Mid Lactation Mastitis In A Mouse Model, Rhonda Nicole Chronis

Theses and Dissertations

Clinical cases of bovine mastitis are most severe in the early stages of lactation. The causes of this increased propensity for severe mastitis during early lactation, compared to mid and late lactation are unclear. In order to better understand the early lactation immune response to mastitis, a murine model of mastitis was employed. Intramammary inoculation of a mastitis causing Escherichia coli strain was performed in lactating mice at various stages of lactation to model the immune response seen in cows during lactation. In our experiments, mice in the early stages of lactation exhibited altered mRNA expression of cytokines IL-1β, IL-6, …

The Role Of Pas Kinase And Cbf1 In Cellular Metabolism, Jenny Adele Pattison, Julianne H. Grose

Journal of Undergraduate Research

The most recent National Institute of Health report concludes that 68.8% of American adults are considered to be overweight or obese1. With this trend in obesity, metabolic diseases such as heart disease, stroke, cancer, and diabetes are on the rise. The critical yet basic function of cellular homeostasis is a central component in controlling these diseases. Sensory protein kinases are essential in the phosphorylation of many protein substrates, allowing them to control several metabolic functions and appropriately allocate glucose, maintaining homeostasis in cells. PAS kinase is a sensory protein kinase that is highly conserved and plays a crucial role in …

Bacteriophages As A Biocontrol Agent Of Soft Rot In Potatoes, Trevor Hodson, Don Breakwell

Journal of Undergraduate Research

Pectobacterium carotovorum (Pcc) is a principle causative agent of soft rot in potatoes. It causes losses of up to 60% in potato yields in the USA (Mantsebo et. Al, 2014) and approximately $50-$100 million dollars’ worth of losses yearly in multiple types of crops. Because potatoes grow beneath the soil surface, there is no way of treating the potatoes until they are harvested. Even then, no postharvest methods of controlling this pathogen exist (Wood et. Al, 2013). A spray-able bacteriophage cocktail specific to Pcc could save millions of dollars-worth of crops each year by minimizing soft rot.

Illuminating Novel Genes For Escherichia Coli Growth In Varying Phosphate Conditions Using Tnseq And Illumina Sequencing, Tanner Robinson, Dr. William R. Mccleary

Journal of Undergraduate Research

The purpose of my project is to identify novel genes in the genome of Escherichia coli ( E coli) associated with phosphate homeostasis under high phosphate conditions. The previous efforts that have been employed were great and innovative for their time, but both lacked the technology currently available, and focused solely on lowphosphate response. Other labs have used variations of the TnSeq protocol that I have, to address a variety of questions in many organisms, with great success (Subashchandrabose et al., 2013).

A Phenome Wide Association Study Of Multiple Sclerosis And Comorbidities, Scott Frodsham, Mary Davis

Journal of Undergraduate Research

Genome wide association studies (GWAS) have identified relationships between many different genes and diseases. GWAS studies scan whole genomes of many individuals and then associate genetic variants with diseases that the individuals have. In contrast, a phenome wide association study (PheWAS), looks at the phenotypes of many individuals and associates those phenotypes with one or more genetic variants. Electronic medical records (EMRs) linked to DNA biobanks provide both clinical and genetic data of patients. This study utilizes one such EMRlinked DNA biobank called BioVU from Vanderbilt University.

Development Of An Animal Model In Which To Study Hiv-2, Taalin Rasmussen, Bradford Berges

Journal of Undergraduate Research

Human Immunodeficiency Virus (HIV) attacks and destroys the cells of the immune system necessary to mounting an effective immune response to successfully fight off infection. Unchecked, an HIV infection leads to a gradual deterioration of the immune system, giving rise to many opportunistic infections not normally seen in people with a functioning immune system. There are two subtypes of HIV: HIV-1, which accounts for the most HIV infections worldwide, and HIV-2, which is most commonly seen in West Africa and countries with strong socioeconomic ties to West Africa, such as France, Portugal, Spain, Angola, Mozambique, and Brazil (1). There is, …

Visualizing Hemocyte Extracellular Traps In Galleria Mellonella, Timothy Siebach, Davidson Erickson

Journal of Undergraduate Research

Neutrophil extracellular traps (NETs) were recently found to be an additional method that neutrophils use to combat pathogens. They are composed of nuclear DNA and proteins that possess anti-microbial properties. Some studies suggest that in addition to killing pathogens, NETs also serve as a barrier to prevent pathogens from spreading. Mammalian infection models are known to possess NETs, but these models often raise ethical issues and can be quite costly. Using an insect such as Galleria mellonella is a valuable alternative. Insects possess an intricate innate immune response comparable to mammals in many ways. There is already substantial evidence indicating …

Membrane On The Frontier, Erin Wu, David Erickson

Journal of Undergraduate Research

Antimicrobial chemokines is a type of host defense protein that play an important role in the natural defenses of many organisms such as defending skin and mucosal tissues against bacterial infections.These peptides have a net positive charge, which contributes to membrane permeabilization and/or cell penetration. It has been observed that CCL28, an antimicrobial chemokine, binds to the surface of bacteria, however, it is not known how they bind, or what they do, to cause bacterial death. Research has demonstrated that Yersinia pseudo tuberculosis with mutations in lipopolysaccharide Oantigen and core synthesis genes (rfaD mutation) exhibit increased binding affinity for the …

Unusual Hprt Expression In Prostate Cancer Cells And Its Impact On Potential Treatment, Michelle H. Townsend, Abi M. Felsted, Michael D. Anderson, Richard A. Robison, Kim L. O'Neill

Biomedical Engineering Western Regional Conference

Unusual HPRT Expression in Prostate Cancer Cells and its Impact on Potential Treatment