Date of Award

2016

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Walstrom, Katherine

Area of Concentration

Chemistry

Abstract

The nematode C. elegans has the capability of entering an alternative developmental pathway in its life cycle known as the dauer larval stage. Entry into this developmental stage is complemented by a metabolic anaerobic shift. C. elegans are triggered to enter this metabolic state due to harsh environmental conditions that include high population density and low food supply. The dauer phase allows C. elegans to live significantly longer than worms following the mainstream developmental pathway (2-3 weeks as opposed to days,). Dauers are also able to enter the L4 stage once environmental stressors subside. Through the use of mass spectroscopy, SODH-1 (a proposed sorbitol dehydrogenase protein,) has been found to be expressed in dauer worms upwards of 3 to 4 times more compared to their L3 worm counterparts. The C. elegans sodh-1 gene was inserted into the expression vector pET303/CT-His. and overexpressed in E. coli Rosetta cells. The gene vector included a 6x Histidine tag at the C-terminus region of the protein, allowing for purification via histidine binding resin chromatography. The purified protein was quantified by a Bradford assay. The purified protein was extensively tested using substrates and additives in kinetics assays to determine its reactivity. While low amounts of activity were detected in the presence of fructose and acetaldehyde, the kinetics assays still need to be optimized further.

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