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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
The objective for Under the Camper Shell was to build a prototype of a full living environment within the confines of a pickup truck bed and camper shell. The total volume available to work with is approximately 85ft3. This full living environment entails functioning systems for essential modern living, providing shelter and spaces for cooking, sleeping, eating, and sanitation. The project proved to be very challenging from the start. First, the livable space is extremely small, being only tall enough for one to sit up straight. The truck and camper shell were both borrowed items, so no modifications were allowed for either, e.g. drilling holes for mounting. The idea was to create a system that could be easily removed, transforming it from a camper to a utility truck. The systems developed for the living environment would be modular and transformative so to accommodate for different necessities when packing. The goal was to create a low-water system with sustainability in mind. Insulating the space was the largest challenge and the most rewarding, using body heat to warm the space and insulate from the elements. Comfort systems were made of high density foam cushions in sections to allow folding and stacking for different functions (sleeping, lounging, and sitting). Sanitation is necessary for healthy living and regular human function. A composting toilet was used for the design, lending to low-water usage and is sustainable over time. Saw dust would be necessary for its function, but upon composting, the unit will generate sufficient amounts of heat to act as a space heater. Showering serves the functions of exfoliation and ridding of bacteria, both of which bath wipes can accomplish, limiting massive volumes of water storage and waste. Storage systems were also designed for modularity. Hooks were installed the length of the bed for hanging or securing items as necessary. Some are available for hanging bags. A cabinetry rail also runs the length of the bed to allow movement of hard storage to accommodate different scenarios. The cooking method is called "sous-vide", a method of cooking food in air-tight bags submerged in hot water. The water is reusable for cooking and no dishes are necessary for serving. Overall, the prototype fulfilled its function as a full living environment with few improvements necessary for future use.
ContributorsLimsirichai, Pimwadee (Author) / Foy, Joseph (Thesis director) / Parrish, Kristen (Committee member) / Barrett, The Honors College (Contributor) / Materials Science and Engineering Program (Contributor) / School of Sustainability (Contributor)
Created2014-12
Description
Gle1 is an mRNP export mediator with major activity localized to the nuclear pore complex in eukaryotic cells. The protein's high preservation across vast phylogenetic distances allows us to approximate research on the properties of yeast Gle1 (yGle1) with those of human Gle1 (hGle1). Research at Vanderbilt University in 2016, which provides the research basis of this thesis, suggests that the coiled-coil domain of yGle1 is best crystallized in dicationic aqueous conditions of pH ~8.0 and 10\u201420% PEG 8000. Further exploration of crystallizable microconditions revealed a favorability toward lower pH and lower PEG concentration. Following the discovery of the protein's native crystallography conditions, a comprehensive meta-analysis of scientific literature on Gle1 was conducted on the association of Gle1 mutations with neuron disease.
ContributorsGaetano, Philip Pasquale (Author) / Foy, Joseph (Thesis director) / Dawson, T. Renee (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12