Barrett, The Honors College Thesis/Creative Project Collection
Barrett, The Honors College at Arizona State University proudly showcases the work of undergraduate honors students by sharing this collection exclusively with the ASU community.
Barrett accepts high performing, academically engaged undergraduate students and works with them in collaboration with all of the other academic units at Arizona State University. All Barrett students complete a thesis or creative project which is an opportunity to explore an intellectual interest and produce an original piece of scholarly research. The thesis or creative project is supervised and defended in front of a faculty committee. Students are able to engage with professors who are nationally recognized in their fields and committed to working with honors students. Completing a Barrett thesis or creative project is an opportunity for undergraduate honors students to contribute to the ASU academic community in a meaningful way.
Displaying 1 - 2 of 2
Description
Cell-free protein synthesis (CFPS) is becoming an increasingly popular method of in vitro protein expression for biotechnology applications. However, there is still no comprehensive resource that outlines the most effective lysate and template combinations for efficient eukaryotic CFPS. To address this issue, expression vectors were constructed and assayed in order to determine their activity within three commercial eukaryotic CFPS systems: Wheat Germ Extract (WGE), Rabbit Reticulocyte Lysate (RRL), and HeLa Cell Lysate (HCL). Previously in the Chaput lab, a luciferase reporter vector was expressed in each lysate system, testing different template variables impacting protein expression, including the 5' UTR sequence, presence of poly(A) tail, and DNA type. It was found that plasmid DNA templates generally yielded ~500-fold greater amount of protein than linear DNA templates and the inclusion of a poly(A) tail did not significantly increase protein expression in the plasmid systems. Additionally, the incorporation of a viral translation enhancing sequence into the 5' UTR increased translation in a lysate-specific manner. The HCL system had a strong preference for the EMCV sequence, WGE had a preference for the sequences from AMV and TMV, and RRL showed no specific preference. Overall, the HCL-EMCV system generated the greatest amount of protein per volume, producing 10-fold more protein than the second best template-lysate combination tested. Here, four human genes fused with a c-Myc tag were expressed in each lysate using the EMCV 5' UTR sequence in order to test the generality of the previous results. Protein synthesis was assayed using a luciferase construct with a c-Myc tag to recapitulate the previous luminometer data and western blotting of the human proteins. These analyses showed the same EMCV expression trends across all systems, with the HCL system synthesizing the greatest amount of each protein. In the future, when choosing commercial eukaryotic CFPS systems for gene expression, these template variables should be considered when performing cost analysis for cell-free protein production.
ContributorsHartsough, Emily Mae (Author) / Chaput, John (Thesis director) / Chandler, Douglas (Committee member) / Larsen, Andrew (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor) / School of Arts, Media and Engineering (Contributor)
Created2015-05
Description
Birds have unusually high plasma glucose concentrations compared to mammals of similar size despite their high metabolic rate. While birds use lipids as their main source of energy, it is still unclear how and why they maintain high plasma glucose concentrations. To investigate a potential underlying mechanism, this study looks at the role of lipolysis in glucose homeostasis. The purpose of this study is to examine the effects of decreased glycerol availability (through inhibition of lipolysis) on plasma glucose concentrations in mourning doves. The hypothesis is that decreased availability of glycerol will result in decreased production of glucose through gluconeogenesis leading to reduced plasma glucose concentrations. In the morning of each experiment, mourning doves were collected at the Arizona State University Tempe campus, and randomized into either a control group (0.9% saline) or experimental group (acipimox, 50mg/kg BM). Blood samples were collected prior to treatment, and at 1, 2, and 3 hours post-treatment. At 3 hours, doves were euthanized, and tissue samples were collected for analysis. Acipimox treatment resulted in significant increases in blood glucose concentrations at 1 and 2 hours post- treatment as well as renal triglyceride concentrations at 3 hours post-treatment. Change in plasma free glycerol between 0h and 3h followed an increasing trend for the acipimox treated animals, and a decreasing trend in the saline treated animals. These results do not support the hypothesis that inhibition of lipolysis should decrease blood glycerol and blood glucose levels. Rather, the effects of acipimox in glucose homeostasis appear to differ significantly between birds and mammals suggesting differing mechanisms for glucose homeostasis.
ContributorsKouteib, Soukaina (Author) / Sweazea, Karen (Thesis director) / Deviche, Pierre (Committee member) / Chandler, Douglas (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2015-05