Matching Items (70)
Description
Heterotrophs such as E. coli contain metabolic pathways with enzymes called carboxylases that are capable of fixing CO2 gas to form metabolites, which has implications for aiding with CO2’s role in climate change. The reductive branch of the tricarboxylic acid (TCA) cycle serves as an important pathway for NAD+ regeneration in enteric bacteria in anaerobic conditions and leads to the production of succinate, a useful industrial product. The enzyme phosphoenolpyruvate carboxykinase is responsible for fixing CO2 in the conversion of PEP to OAA within this pathway and has potential to be a significant carbon fixation module in heterotrophic organisms. This project explored pck genes from select organisms by transforming plasmids to test if these variants have improved kinetics compared to the native E. coli Pck and to investigate their ability to improve succinate bioproduction.
ContributorsModukuri, Shree (Author) / Wang, Xuan (Thesis director) / Godar, Amanda (Committee member) / Barrett, The Honors College (Contributor) / School of Molecular Sciences (Contributor) / School of International Letters and Cultures (Contributor)
Created2024-05
Description
The symbiosis between termites and their parabasalid hindgut protists centers around the wood digestion that is needed for both species to acquire the nutrients from wood. One of the important carbohydrate-active proteins required for the wood breakdown are glycoside hydrolase (GH) families. Previous studies have looked at the phylogeny of some of these protein families from a termite whole gut transcriptome or in a different context than lignocellulose digestion. In this study, we attempt to understand the function and evolution of these GH families in the context of protist evolution by using protist single cell transcriptomes. 14 families of interest were chosen to create phylogenetic trees: GH2, GH3, GH5, GH7, GH8, GH9, GH10, GH11, GH26, GH43, GH45, GH55, GH67, GH95 for their interesting expressions across different protists such as being present in all protists or being present in only termite-associated protists. The dbCAN2 (automated Carbohydrate-active enzyme ANnotation) program was used to find GH families in each of the protist single cell transcriptomes and additional characterized sequences registered on the National Center for Biotechnology Information to create phylogenetic trees for each of the GH families of interest. Results show that many of the GH families expressed in protists were acquired through horizontal gene transfer from fungi and bacteria. Additionally, comparison to the parabasalid phylogeny indicates most GH families evolved independently from the protists. Based on the pattern of expression of these GH families throughout different protist orders, conclusions can be made about whether the specific family was vertically or horizontally acquired in the termite symbionts.
ContributorsJahan, Israa (Author) / Gile, Gillian (Thesis director) / Wang, Xuan (Committee member) / Swichtenberg, Kali (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor) / Department of Psychology (Contributor)
Created2023-05
Description
Current industrial production of petrochemicals releases CO2 as a byproduct into the
atmosphere, contributing to climate change. The sustainable alternative, microbial carbon
capture, has primarily focused on phototrophs that have naturally occurring carbon fixation
pathways, but are slow-growing, difficult to genetically engineer, and require sunlight, which
limits their large-scale production capacity. Using a heterotroph such as Escherichia coli allows for chemical production at high titers, rates, and yields (TRY) while being fast growing and easy to genetically engineer. Under fermentation conditions, the carboxylases in E. coli fix inorganic carbon in the reductive branch of the TCA cycle, producing industrially relevant chemical precursors such as succinate. However, the carboxylase’s access to CO2 is limited by the conditions surrounding it; most of the inorganic carbon inside the cell is in the form of
bicarbonate. Increasing the local concentration of CO2 near the carboxylase may improve the
kinetics of the pathway. To do this, a fusion protein that colocalizes carbonic anhydrase and
phosphoenolpyruvate carboxykinase (Pck) was created. However, since strains expressing this fusion protein did not grow above OD600 = 1 under fermentation conditions, further design optimization and investigation is needed.
ContributorsConway, Dalton (Author) / Nielsen, David (Thesis director) / Wang, Xuan (Committee member) / Barrett, The Honors College (Contributor) / Chemical Engineering Program (Contributor) / School of Mathematical and Statistical Sciences (Contributor)
Created2024-05
Description
With dwindling water resources due to drought and other pressures, water utilities are seeking to tap into alternative water sources as a means to improve water sustainability. Reclaimed water consists of treated wastewater and is widely used for non-potable purposes, such as irrigation, both agricultural and recreational. However, the reclaimed water distribution system can be subject to substantial regrowth of microorganisms, including opportunistic pathogens, even following rigorous disinfection. Factors that can influence regrowth include temperature, organic carbon levels, disinfectant type, and the time transported (i.e., water age) in the system. One opportunistic pathogen (OP) that is critical to understanding microbial activity in both reclaimed and drinking water distribution systems is Acanthamoeba. In order to better understand the potential for this amoeba to proliferate in reclaimed water systems and influence other OPs, a simulated reclaimed water distribution system was studied. The objective of this study was to compare the prevalence of Acanthamoeba and one of its endosymbionts, Legionella, across varying assimilable organic carbon (AOC) levels, temperatures, disinfectants, and water ages in a simulated reclaimed water distribution system. The results of the study showed that cooler temperatures, larger water age, and chlorine conditions yielded the lowest detection of Acanthamoeba gene copies per mL or cm2 for bulk water and biofilm samples, respectively.
ContributorsDonaldson, Kandace (Author) / Ankeny, Casey (Thesis director) / Edwards, Marc (Committee member) / Pruden, Amy (Committee member) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12
Description
Translating research has been a goal of the Department of Health and Human Services since 1999. Through two years of iteration and interview with our community members, we have collected insights into the barriers to accomplishing this goal. Liberating Science is a think-tank of researchers and scientists who seek to create a more transparent process to accelerate innovation starting with behavioral health research.
ContributorsRaghani, Pooja Sioux (Author) / Hekler, Eric (Thesis director) / Buman, Matthew (Committee member) / Pruthi, Virgilia Kaur (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / Biomedical Informatics Program (Contributor)
Created2014-05
Description
Methane (CH4) is very important in the environment as it is a greenhouse gas and important for the degradation of organic matter. During the last 200 years the atmospheric concentration of CH4 has tripled. Methanogens are methane-producing microbes from the Archaea domain that complete the final step in breaking down organic matter to generate methane through a process called methanogenesis. They contribute to about 74% of the CH4 present on the Earth's atmosphere, producing 1 billion tons of methane annually. The purpose of this work is to generate a preliminary metabolic reconstruction model of two methanogens: Methanoregula boonei 6A8 and Methanosphaerula palustris E1-9c. M. boonei and M. palustris are part of the Methanomicrobiales order and perform hydrogenotrophic methanogenesis, which means that they reduce CO2 to CH4 by using H2 as their major electron donor. Metabolic models are frameworks for understanding a cell as a system and they provide the means to assess the changes in gene regulation in response in various environmental and physiological constraints. The Pathway-Tools software v16 was used to generate these draft models. The models were manually curated using literature searches, the KEGG database and homology methods with the Methanosarcina acetivorans strain, the closest methanogen strain with a nearly complete metabolic reconstruction. These preliminary models attempt to complete the pathways required for amino acid biosynthesis, methanogenesis, and major cofactors related to methanogenesis. The M. boonei reconstruction currently includes 99 pathways and has 82% of its reactions completed, while the M. palustris reconstruction includes 102 pathways and has 89% of its reactions completed.
ContributorsMahendra, Divya (Author) / Cadillo-Quiroz, Hinsby (Thesis director) / Wang, Xuan (Committee member) / Stout, Valerie (Committee member) / Barrett, The Honors College (Contributor) / Computing and Informatics Program (Contributor) / School of Life Sciences (Contributor) / Biomedical Informatics Program (Contributor)
Created2014-05
Description
This study focused on the connection between the EnvZ/OmpR two-component regulatory system and the iron homeostasis system in Escherichia coli, specifically how a mutant form of EnvZ11/OmpR is able to reduce the expression of fepA::lacZ, a reporter gene fusion in E. coli. FepA is one of several outer membrane siderophore receptors that allow extracellular siderophores bound to iron to enter the cells to power various biological processes. Previous studies have shown that in E. coli cells that expressed a mutant allele of envZ, called envZ11, which led to altered expression of various iron genes including down regulation of fepA::lacZ. The wild type EnvZ/OmpR system is not considered to regulate iron genes, but because these envz11 strains had downregulated fepA::lacZ, this study was undertaken to understand the connection and mechanisms of this downregulation. A large number of Lac+ revertants were obtained from the B32-2483 strain (envz11 and fepA::lacZ) and 7 Lac+ revertants that had reversion mutations not directly correcting the envZ11 allele were further characterized. With P1 phage transduction genetic mapping that involved moving a kanamycin resistance marker linked to fepA::lacZ, two Lac+ revertants were found to have their reversion mutations in the fepA promoter region, while the other five revertants had their mutations mapping outside the fepA region. These two revertants underwent DNA sequencing and found to carry two different single base pair mutations in two different locations of the fepA promoter region. Each one is in the Fur repressor binding region, but one also may have affected the Shine-Dalgarno region involved in translation initiation. All 7 reveratants underwent beta-galactosidase assays to measure fepA::lacZ expression. The two revertants that had mutations in the fepA promoter region had significantly increased fepA activity, with the revertant with the Shine-Dalgarno mutation having the most elevated fepA expression. The other 5 revertants that did not map in the fepA region had fepA expression elevated to the same level as that found in the wild type EnvZ/OmpR background. The data suggest that the negative effect of envZ11 can be overcome by multiple mechanisms, including directly correcting the envZ11 allele or changing the fepA promoter region.
ContributorsKalinkin, Victor Arkady (Co-author) / Misra, Rajeev (Co-author, Thesis director) / Mason, Hugh (Committee member) / Foy, Joseph (Committee member) / Biomedical Informatics Program (Contributor) / School of Life Sciences (Contributor) / W. P. Carey School of Business (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
Many bacteria actively import environmental DNA and incorporate it into their genomes. This behavior, referred to as transformation, has been described in many species from diverse taxonomic backgrounds. Transformation is expected to carry some selective advantages similar to those postulated for meiotic sex in eukaryotes. However, the accumulation of loss-of-function alleles at transformation loci and an increased mutational load from recombining with DNA from dead cells create additional costs to transformation. These costs have been shown to outweigh many of the benefits of recombination under a variety of likely parameters. We investigate an additional proposed benefit of sexual recombination, the Red Queen hypothesis, as it relates to bacterial transformation. Here we describe a computational model showing that host-pathogen coevolution may provide a large selective benefit to transformation and allow transforming cells to invade an environment dominated by otherwise equal non-transformers. Furthermore, we observe that host-pathogen dynamics cause the selection pressure on transformation to vary extensively in time, explaining the tight regulation and wide variety of rates observed in naturally competent bacteria. Host-pathogen dynamics may explain the evolution and maintenance of natural competence despite its associated costs.
ContributorsPalmer, Nathan David (Author) / Cartwright, Reed (Thesis director) / Wang, Xuan (Committee member) / Sievert, Chris (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
The purpose of this study is to analyze the stereotypes surrounding four wind instruments (flutes, oboes, clarinets, and saxophones), and the ways in which those stereotypes propagate through various levels of musical professionalism in Western culture. In order to determine what these stereotypes might entail, several thousand social media and blog posts were analyzed, and direct quotations detailing the perceived stereotypical personality profiles for each of the four instruments were collected. From these, the three most commonly mentioned characteristics were isolated for each of the instrument groups as follows: female gender, femininity, and giggliness for flutists, intelligence, studiousness, and demographics (specifically being an Asian male) for clarinetists, quirkiness, eccentricity, and being seen as a misfit for oboists, and overconfidence, attention-seeking behavior, and coolness for saxophonists. From these traits, a survey was drafted which asked participating college-aged musicians various multiple choice, opinion scale, and short-answer questions that gathered how much they agree or disagree with each trait describing the instrument from which it was derived. Their responses were then analyzed to determine how much correlation existed between the researched characteristics and the opinions of modern musicians. From these results, it was determined that 75% of the traits that were isolated for a particular instrument were, in fact, recognized as being true in the survey data, demonstrating that the stereotypes do exist and seem to be widely recognizable across many age groups, locations, and levels of musical skill. Further, 89% of participants admitted that the instrument they play has a certain stereotype associated with it, but only 38% of people identify with that profile. Overall, it was concluded that stereotypes, which are overwhelmingly negative and gendered by nature, are indeed propagated, but musicians do not appear to want to identify with them, and they reflect a more archaic and immature sense that does not correlate to the trends observed in modern, professional music.
ContributorsAllison, Lauren Nicole (Author) / Bhattacharjya, Nilanjana (Thesis director) / Ankeny, Casey (Committee member) / School of Life Sciences (Contributor) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
This project aims to address the current protocol regarding the diagnosis and treatment of traumatic brain injury (TBI) in medical industries around the world. Although there are various methods used to qualitatively determine if TBI has occurred to a patient, this study attempts to aid in the creation of a system for quantitative measurement of TBI and its relative magnitude. Through a method of artificial evolution/selection called phage display, an antibody that binds highly specifically to a post-TBI upregulated brain chondroitin sulfate proteoglycan called neurocan has been identified. As TG1 Escheria Coli bacteria were infected with KM13 helper phage and M13 filamentous phage in conjunction, monovalent display of antibody fragments (ScFv) was performed. The ScFv bind directly to the neurocan and from screening, phage that produced ScFv's with higher affinity and specificity to neurocan were separated and purified. Future research aims to improve the ScFv characteristics through increased screening toward neurocan. The identification of a highly specific antibody could lead to improved targeting of neurocan post-TBI in-vivo, aiding researchers in quantitatively defining TBI by visualizing its magnitude.
ContributorsSeelig, Timothy Scott (Author) / Stabenfeldt, Sarah (Thesis director) / Ankeny, Casey (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2015-05