Office of Undergraduate Research

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Emphasizing equitable and inclusive access to research opportunities, the University of Maryland's Office of Undergraduate Research (OUR) empowers undergraduates and faculty to engage and succeed in inquiry, creative activity, and scholarship. This collection includes materials shared by undergraduate researchers during OUR events. It also encompasses materials from Undergraduate Research Day 2020, Undergraduate Research Day 2021, and Undergraduate Research Day 2022, which were organized by the Maryland Center for Undergraduate Research.

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    INVESTIGATING THE ROLE OF E. COLI TCA CYCLE METABOLISM IN BACTERIOPHAGE REPLICATION
    (2024) Kavalov, Lilith; Weaver, Trinity; O'Hara, Jessica
    Bacteriophages are viruses that specifically infect and hijack Escherichia coli's metabolic processes to proliferate, ultimately destroying the host cell in the process. The Tricarboxylic Acid Cycle (TCA Cycle) is a multi-step aerobic enzyme-catalyzed pathway that occurs in the cytoplasm of E. coli and is responsible for generating the electron-carrier molecules NADH and FADH2 which are crucial for generating ATP for the cell in future steps of cellular respiration. The E. coli genes, acnB, and acnA, encode enzymes that catalyze different reactions that are necessary for the TCA cycle. We hypothesize that the removal of these genes would negatively impact the growth rate and ATP levels of E. coli and, as a result, inhibit or slow the replication of bacteriophage. To determine the effects of the removal of these genes, enzyme assays, comparative growth curves of the knockout strains, and plaque assays of bacteriophage replication were measured and investigated. Furthermore, we quantified the knockout’s effects by collecting lysis curves as well as performing an ATP assay using bioluminescence.
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    Decreased Host-Cell ATP Levels Affects Bacteriophage Replication in Knockout E. coli Strains
    (2024) Alumyar, Marrie; Beitzell, Lauren; Bradish, Kristin; Kato, Rion; Vozna, Alina; O'Hara, Jessica
    Bacteriophages are viruses that use host cell metabolic resources for replication. Altering Escherichia coli's ATP production pathway can inhibit bacteriophage replication, offering a new approach to bacteriophage therapy.The atp genes encode ATP synthase subunits crucial for ATP generation in E. coli. Knockouts ΔatpA, B, D, E, and H, alongside the parent strain, were studied. Focus narrowed to ΔatpA and B due to significant deviations from the parent strain. It is hypothesized that these knockout strains reduce growth in E. coli and bacteriophage due to decreased ATP production, vital for metabolism and phage replication. Comparative growth assays of E. coli parent and ATP knockout strains were conducted in LB-rich media and M9 minimal media. T4 bacteriophage replication was measured through lysis curves, plaque assays, and two time-point phage titer experiments, chosen for consistent replication. Characterization of T4 bacteriophage replication revealed ΔatpA's crucial role, showing difficulties in growth and lysing. ΔatpA required 10-4 dilutions in phage titer experiments due to low PFU/mL, contrasting with 10-7 dilutions for other strains. ATP assay data showed significantly lower ATP concentration (319nM) in ΔatpB compared to the parent strain, also implying its crucial role in ATP synthesis.Future research will focus on characterizing phage replication in ATP synthase knockout strains using E. coli ATP synthase inhibitors to deepen understanding of phage-host interactions. Controlled bacteriophage manipulation can be studied further to have a better understanding of the application of bacteriophage therapy and to potentially improve its clinical efficacy.