Purifying Scorpion Antarease from E. coli
Hudson, Victoria Elizabeth
Vesicular trafficking is a system of intracellular transport that allows for the distribution and secretion of molecules. In the pancreas, normal secretory processes have been found to be interrupted by Antarease, a metalloprotease isolated from the venom of the Brazilian scorpion Tityus serrulatus. Antarease cleaves membrane proteins that take part in the transport and release of intracellular vesicles. The purpose of this project is to develop and optimize a production system for Antarease to provide higher concentrations of protein. Antarease was cloned into a cytoplasmic expression system and potential candidates were screened for protein expression. After identifying Antarease-expressing candidates, SDS-PAGE and Western Blotting were used to confirm this identification and determine relative concentration of Antarease in a small scale system. Once small scale growth conditions were determined, larger scale protein inductions were carried out and evaluated. It was found that the small scale conditions did not translate well to a larger scale. However, protein is still being expressed and minor optimization should allow higher level expression in a scaled up system.
Hudson, Victoria Elizabeth. (May 2018). Purifying Scorpion Antarease from E. coli (Honors Thesis, East Carolina University). Retrieved from the Scholarship. (http://hdl.handle.net/10342/6872.)
Hudson, Victoria Elizabeth. Purifying Scorpion Antarease from E. coli. Honors Thesis. East Carolina University, May 2018. The Scholarship. http://hdl.handle.net/10342/6872. September 23, 2020.
Hudson, Victoria Elizabeth, “Purifying Scorpion Antarease from E. coli” (Honors Thesis., East Carolina University, May 2018).
Hudson, Victoria Elizabeth. Purifying Scorpion Antarease from E. coli [Honors Thesis]. Greenville, NC: East Carolina University; May 2018.
East Carolina University