Microbiology Commons^™

Engineering A Pathogen Specific High Affinity T-Cell Receptor Called Llo56, John Hancock, Dr. Scott Weber

Journal of Undergraduate Research

CD4+ helper T-cells play a vital role in the body’s immune response. When infectious agents attack the body, phagocytes engulf these invaders and present a peptide segment of the pathogen on a receptor (called MHCII). These receptors are located on the surface of the cell and the displayed peptide is termed an epitope. CD4+ T-cells with T-cell receptors (TCRs) specific for the displayed peptide bind to the MHCII complex. It is this binding that releases chemical signals to initiate an immune response. A disadvantage of TCRs is that their wild-type affinity for MHC is low and that they are membrane …

Discovery And Characterization Of Bacteriophage To Control Mrsa, Jacob B. Hatch, Bradford Berges

Journal of Undergraduate Research

About 20% of humans are carriers of Staphylococcus aureus (SA). There were an estimated 11,000 deaths in the United States in 2005 attributed to SA, with the majority caused by MRSA (Methicillin Resistant Staphylococcus Aureus) isolates [1]. Many MRSA isolates have developed resistance to all but one antibiotic drug: vancomycin. However, other bacteria have developed resistance to vancomycin, suggesting that in time MRSA will likewise become non-responsive to this last available drug and MRSA infections will be untreatable. This project looks to find an alternate method of MRSA treatment. Bacteriophage (phage) are viruses that infect bacterial cells in order …

Use Of Humanized Mice To Study The Production Of Human Antibodies, Steven Hallam, Dr. Bradford Berges

Journal of Undergraduate Research

The human immune system is composed of two main subsets: innate and adaptive immunities. Among the adaptive immune cells, B cells play a vital role in stopping infectious agents through the production of antigen specific antibodies. Human B cells are important targets of infection for many human-specific viruses that are poorly understood. Since the infection and study of human subjects is not possible, animal models have been developed to mimic the human immune environment. Rag2-/-γc-/- mice engrafted with human hematopoietic stem cells produce human B lymphocytes and these cells are capable of mounting antigen-specific human antibody responses. However, there is …

Mapping Interaction Through Compensatory Phou Mutants In Escherichia Coli, Tanner Dean, William Mccleary

Journal of Undergraduate Research

The purpose of this experiment was to identify the physical interaction site between two proteins in Escherichia Coli. These two proteins, PhoU and PhoR, are two proteins involved in Phosphate signal transduction. When these two proteins are mutated, so they no longer interact, the pathway is broken and signaling is inhibited. To break this pathway, we mutated one protein (PhoR) until we developed several mutations that no longer interact with PhoU. This loss of interaction is caused by a physical change to the site of interaction between the two proteins. We hypothesized that we could find a naturally occurring PhoU …

Fire Blight Treatment With Lytic Bacteriophage, Todd Jarvis, Julianne Grose

Journal of Undergraduate Research

Fire Blight, caused by the bacteria Erwinia amylovora, is a highly contagious fruit tree disease that is difficult to treat. This disease causes a tree to whither away, as if it had been burned. It is estimated that Fire Blight causes more than a 100 million dollar loss in agricultural goods each year¹. In an attempt to treat fire blight and prevent it from spreading, large doses of antibiotics are sprayed on infected trees. These antibiotics do little in the way of helping to treat the trees. In addition, the antibiotic run off from the treatment finds …

Lymphoblastoid Cell Line Generation From Sle Patient Lymphocytes, Vera Mayhew, Brian Poole

Journal of Undergraduate Research

Epstein-Barr Virus (EBV) is one of the viruses found most frequently in the human population. Approximately 95% of the population is infected in the United Sates by the age of 40. When primary infection occurs is young adulthood approximately 50% of cases result in infectious mononucleosis. Whether EBV infection results in mononucleosis manifestations or not, the virus stays latent in the infected individual’s B cells for the rest of their life.

How Chemokine Ccl28 Binds To Yersinia Pseudotuberculosis, Jeffrey Moffat, David Erickson

Journal of Undergraduate Research

Chemokines are tiny proteins that play a valuable role in defending our bodies against bacteria and other pathogens. Chemokines are best known for their role in attracting immune cells to areas of infection. Some chemokines, however, have demonstrated the ability to not only recognize pathogens and/or recruit white blood cells, but also to kill various classes of bacteria and fungi¹. One of these antimicrobial chemokines is CCL28.

P. Larvae Bacteriophages As A Treatment For American Foulbrood In Honeybees, Bryan Merrill, Sandra Burnett

Journal of Undergraduate Research

The deadly honey bee disease American Foulbrood (AFB) is caused by the bacterium Paenibacillus larvae. The widespread use of antibiotics to prevent or treat AFB has led to many drug-resistant strains of P. larvae. Bacteriophages can target and destroy specific strains of bacteria and can be used to treat bacterial infections. This study accomplished two main goals. We determined the efficacy of previously isolated P. larvae phages as a treatment for AFB by applying a phage spray to infected beehives. We also isolated P. larvae bacteria from the infected hives we treated and identified phages to better counteract …

Effect Of Ebi2 On B Cell Migration, Jeffrey Mella, Brian Poole

Journal of Undergraduate Research

My mentored research project was a terrific experience. I have been mentored with Dr. Brian Poole for the last three years. As a result of the project that was made possible by the ORCA grant, I was the first author on an abstract submitted and accepted by the International Immunology Summit. This last October I attended the International Immunology Summit in Las Vegas, Nevada. Dr. Poole and I presented our research and I responded to questions from the audience. I am currently an author on a manuscript that in progress that has been submitted for review. I have enjoyed my …

Histone Modifications And Their Effects On Nucleosome Positioning And Gene Expression, Jordon Ritchie, Steven Johnson

Journal of Undergraduate Research

Nucleosome positioning plays an important role in gene regulation and expression. Nucleosomes consist of DNA-histone interactions that comprise the first order of DNA compaction into chromatin in the cell. Modifications to the histone in the nucleosome have been hypothesized to influence the location of the nucleosome on the DNA and therefore the regulation of the gene the nucleosome is forming on. In our original proposal, we proposed to show the effects that different modifications had on the position of the nucleosome on the DNA and the DNA sequence that had the highest affinity for nucleosome formation given a specific histone …

Analysis Of The Adjuvant Effect Of Vitamin D3 In Enhancing Mucosal Immune Responses, Cameron Sargent, Dr. Eric Wilson

Journal of Undergraduate Research

Because skin provides an effective physical barrier, most pathogens that invade the body do so at mucosal sites like the nose and mouth. As such, finding methods of improving mucosal immunity is key to providing greater defense against communicable diseases, the cause of approximately one-quarter of all deaths worldwide [1]. Improving vaccination, the technique in which the body is exposed to a weakened or nonvirulent dosage of a pathogen or toxin to promote an immune response specific to that foreign material, is a potential way of enhancing mucosal immunity. Vaccination has been used for several decades in medicine, nearly eradicating …

How Yersinia Pseudotuberculosis Biofilm Formation Is Regulated By Global Regulator Csra, Jeffrey Schachterle, David Erickson

Journal of Undergraduate Research

The bacteria Yersinia pestis is the causative agent of bubonic plague, and its ability to form biofilm in fleas is essential for plague transmission by fleas [1]. Y. pestis recently evolved from Y. pseudotuberculosis (Y. pstb), and the two have nearly identical genomes. Y. pestis forms a biofilm in fleas that is capable of blocking the flea’s feeding. These biofilms are made up of a community of bacteria adhering to a polysaccharide matrix that is made by the hmsHFRS gene products. Y. pstb is capable of infecting fleas, but will not block the flea’s feeding [1]. Since the …

A Method Of Introducing Site Specific Mutations Into The Phor Gene Of The E. Coli Chromosome, Cameron Woodward, William Mccleary

Journal of Undergraduate Research

Phosphate intake by Escherichia coli serves as a good model system to study phosphate intake by other bacteria. The pho phosphate uptake system used by E. coli is well conserved in pathogens like Vibrio cholerae (1). To better understand this important mode of phosphate uptake a study was performed to examine the interactions between the phoR and phoU proteins of this system. PhoU interacts with the pstSCAB complex, which brings phosphate into the cell. After communicating with pstSCAB the phoU protein interacts with phoR to tell it if phosphate is present in the environment or not. PhoR has been shown …

Characterizing Dna2 In Model Plant Arabidopsis, Connor Woodward, Brent Nielsen

Journal of Undergraduate Research

Dna2 is a protein that is known to be essential for DNA replication in human and yeast mitochondria. In these organisms it acts as both a helicase and an endonuclease. In the model plant Arabidopsis thaliana an ortholog of Dna2 exists but has never been studied. Two questions were answered in this study. The first question is “What would happen to A. thaliana plants that lacked Dna2?”. The second is “Where is Dna2 localized in plant cells?”

Role Of Epstein-Barr Virus Induced Gene Ii (Ebi2) In Epstein-Barr Virus Infection And B-Cell Chemotaxis, 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 was used to publish three peer-reviewed journal articles:

Post-Translational Modifications Of Thymidine Kinase 1, Dr. Kim O'Neill

Journal of Undergraduate Research

Thymidine kinase 1 (TK1) is one diagnostic and prognostic marker which has shown promising potential in a variety of solid tumors and hematological malignancies. Unfortunately there have been challenges associated with developing a clinically useful immunoassay which have prevented TK1 from being fully utilized in the clinical setting at present. One of the major goals of this study was to produce a dependable, reproducible, immunoassay to detect TK1 in serum from cancer patients. There have been many attempts at detecting protein or genetic material in the serum or sputum of lung cancer patients but these all have failed due to …

Elucidating A Precise Role For The Small Heat Shock Proteins Cryab And Hspb2 In Cardiac Robustness, Julianne H. Grose

Journal of Undergraduate Research

Evaluation of how well the academic objectives of the proposal were met

• Specific Aim 1: Perform yeast two-hybrid screens on wild type CryAB, R120G CryAB, and HspB2 in order to create a protein-protein interaction network and identify potential substrates.
• Achievements: Large-scale yeast-two hybrid screens were performed on wild type CryAB, R120G CryAB and HspB2 in order to identify their protein-protein interaction networks and to identify putative substrates.
• Specific Aim 2: Validate the protein-protein interaction network via Y2H dependency tests (A), affinity purification from mammalian cells (B) and chaperone activity assays (C).
• Achievements: Y2H dependency tests were performed for all three …

Validation Of An Nbmp2 Mutant Mouse Model, Laura C. Bridgewater

Journal of Undergraduate Research

Work in the Bridgewater Lab over the last several years has focused on elucidating the function of a novel protein we discovered, nBmp2. The initial discovery presenting work performed in cultured cells (rather than whole animals) was published back in 2010 and we subsequently constructed a mutant mouse line in which nBmp2 is disrupted. Much evidence was accumulating to show that the mice have problems with muscle function, neurological function, and immune function, but some fundamental experiments necessary to validate the mouse model had been giving us problems. The absence of that data prevented us from publishing any reports on …

Tissue Specific Isolation Of Nucleosomes In Caenorhabditis Elegans, Hyrum Shumway, Dr. Steven Johnson

Journal of Undergraduate Research

Hershey and Chase proved DNA to be the genetic material of life in 1952. The structure of DNA largely eluded investigators until Watson and Crick discovered it a year later. These two landmark experiments laid a foundation so that questions in transcription, translation, gene regulation, and inheritance could be answered.

Knockout Nbmp2 Mouse Embryonic Fibroblast Cell Line, Josh Yates, Dr. Laura Bridgewater

Journal of Undergraduate Research

Bone morphogenetic proteins (BMPs) contribute to many different aspects of development including mesoderm formation, heart development, neurogenesis, skeletal development, and axis formation. They have previously been recognized only as secreted growth factors. Recently, researchers in the Bridgewater lab discovered a nuclear variant of Bone Morphogenic Protein 2 (nBMP2). To better understand the function of nBMP2, our lab generated a mutant mouse line in which nBMP2 is missing from the nucleus. These mice display learning and memory deficits, muscle dysfunction, and abnormal immune responses. In order to validate the mouse model and to study the molecular mechanisms of nBMP2 we have …

Inhibition Of Epstein-Barr Virus Infection By Kalanchoe Pinnata Extract, Lance A. Stutz, Dr. Brian Poole

Journal of Undergraduate Research

Epstein-Barr virus (EBV) is a common infectious agent which infects more than 95% of adults worldwide (1). EBV is related to a number of diseases, including cancers such as Burkitt’s lymphoma and nasopharyngeal carcinoma, and is the causative agent of infectious mononucleosis (2). Upon infection, EBV invades antibody-producing immune cells known as B cells. Most people do not show signs or symptoms of the infection because EBV tends to hide in B cells in a latent state. Latent EBV infection contributes to the development of EBV-associated cancers. However, EBV can also be in an active or lytic state. In this …

Ebi2 Expression In Different Latency Stages And Primary B Cells, Jillian Markham, Dr. Brian Poole

Journal of Undergraduate Research

Epstein-­‐Barr Virus (EBV) causes infectious mononucleosis upon primary infection, commonly known as “mono.” Less commonly known is that EBV doesn’t get eradicated from your body after you recover from mono. EBV quietly occupies a small portion of B cells in 90% of human adults [1]. Besides mononucleosis, EBV infection is linked to cancers, lymphomas, and several autoimmune diseases [2.3].

The Effects Of Dna Methylation On Nucleosome Positioning, Marcus Vranes, Dr. Steven Johnson

Journal of Undergraduate Research

Recent studies have attempted to discover the correlation that exists between DNA methylation and nucleosome positioning, but none have explored the direct effect of DNA methylation on nucleosome formation and positioning. This research is directly testing the effects of DNA methylation on nucleosome positioning, and whether the histone octamer has preferred sequences to which it binds, which will add to our understanding of gene expression and regulation. A better understanding of these concepts will help to aid efforts in gene therapy to improve the quality of life for many people who suffer from various genetic conditions.

Biofilm Regulation In Yersinia Pseudotuberculosis Through Elements Downstream Of The Bara/Uvry Two-Component System, Jeffrey Schachterle, Dr. David Erickson

Journal of Undergraduate Research

The bacteria Yersinia pestis is the causative agent of bubonic plague, and its ability to form biofilm in fleas is essential for plague transmission by fleas [1]. Y. pestis recently evolved from Y. pseudotuberculosis (Y. pstb), and the two have nearly identical genomes. Y. pestis forms a biofilm in fleas that is capable of blocking the flea’s feeding. These biofilms are made up of bacteria stuck to a polysaccharide matrix that is made by the hmsHFRS gene products. Y. pstb is capable of infecting fleas, but will block the flea’s feeding [1]. Since the hmsHFRS genes themselves are identical, this …

Detecting Endogenous Ccl28, Sara Mason, Dr. Eric Wilson

Journal of Undergraduate Research

Chemokines play a well-characterized and significant role in the immune system as chemoattractants for white blood cells. Some chemokines have also been shown to have antimicrobial properties in addition to their chemoattractant functions. CCL28 is a chemokine that aids in the recruitment of antibody secreting cells to mucosal surfaces of the body, especially in the large intestine and lactating mammary glands.1 Recent research has shown that, in addition to recruiting lymphocytes, CCL28 also exhibits antimicrobial activity against gram positive and gram negative bacteria, and antifungal activity.2

Development Of A Traveler’S Vaccine Against Dengue Infection, Arthur Lee, Dr. Bradford Berges

Journal of Undergraduate Research

There are currently no vaccines available against dengue virus. Each year, about 400 million people become infected and half a million of them, especially children, develop complications such as dengue hemorrhagic fever or dengue shock syndrome.

Genome Annotation Of Novel Viruses And Discovery Of Critical Sequences In Genes Via Comparative Analysis Of B4 Mycobacteriophage, Cameron Sargent, Dr. Sandra Burnett

Journal of Undergraduate Research

Over the past few years at Brigham Young University and other colleges nationwide, members of the Phage Hunters program have endeavored to find and analyze novel mycobacteriophage in an attempt to create new treatment and research methods for the pathogen Mycobacterium tuberculosis

Comparative Genome Analysis Of Novel Paenibacillus Larvae Bacteriophage, Bryan Merrill, Dr. Sandra Burnett

Journal of Undergraduate Research

Honey bees pollinate one third of the crops in the U.S. diet. The spore-forming bacterium Paenibacillus larvae causes American Foulbrood (AFB), a highly contagious disease that is lethal in honey bee larvae. P. larvae is the most serious pathogen affecting honey bees. The spores can be found in any healthy beehive, are stable for 40 years or more, and cannot be killed by antibiotics. The disease manifests itself when worker bees feed P. larvae spores to larvae and the spores germinate. The bacteria then consume the larvae and turns back into a spore that the worker bees then transfer throughout …

Determining Gene Expression Levels In Healthy And Degraded Alpaca Spinal Discs, Loyd Christensen, Dr. Laura C. Bridgewater

Journal of Undergraduate Research

Intervertebral Disc Degeneration (IDD) is a disease estimated to affect between 60 and 90 percent of the total population at some point in life. Evidence of spinal disc degeneration has been shown in 30 percent of individuals under 40 years old, with earliest symptoms visible at 21 years of age; its prevalence only increases with age. Symptoms of spinal disc degeneration account for more than 13 million doctor visits each year and result in costs of more than $200 billion in lost productivity and health-care related expenses in the US alone. Unfortunately, despite these staggering costs, no widely effective treatment …

Understanding The Irf5 Gene Through Characterization Of Its Four Promoters, Lissenya Argueta, Dr. Brian Poole

Journal of Undergraduate Research

Interferon Regulatory Factor 5 (IRF5) is a transcription factor involved with the innate antiviral immune response and primarily expressed in immune cells1. This protein is responsible for the regulation of interferon activity and cytokine signaling, namely that of pro-inflammatory cytokines. These processes play major roles in autoimmune diseases. IRF5 is also involved with control of cell cycle and apoptosis upon activation2, therefore in certain cancers it can be inactivated3,4. The IRF5 gene has four different promoters for each first exon: 1A, 1B, 1C, and 1D. Each promoter is distinct, displaying unique correlations with certain proteins. However, the resulting IRF5 protein …