Date of Award

6-2014

Document Type

Open Access

Degree Name

Bachelor of Science

Department

Biochemistry

First Advisor

Kristen Fox

Language

English

Keywords

plants, fungi, protists, cell death, proteins

Abstract

Metacaspases are caspase homologs that are found in plants, fungi, and protists. They are involved in programmed cell death, a structured and regulated process to break down cells. There is limited information available on metacaspases and further research is necessary to understand how they work. One reason metacaspases are studied is that they are considered to be a potential drug target for pathogenic microorganisms. Five metacaspases scp1-5 were identified in S. commune through their similarity to the yeast metacaspase Yca1. The overall goal of the lab is to study and characterize these proteins. This thesis outlines the path taken to express the Schizophyllum commune metacaspase scp3 in yeast. Previous efforts in our lab to express metacaspases in E. coli have not been successful. I hypothesized that yeast would be a more desirable expression vehicle due to its closer relationship to S. commune; a more similar environment could yield more active protein. Scp3 has been successfully cloned into a yeast expression vector pYES2.1/V5-His-TOPO® and β-galactosidase expression has been obtained in yeast with the expression control vector pYES1.2/V5-His/lacZ. Future work on this project will be to perform a point mutation in the expression plasmid to remove a stop codon before the scp3 sequence.

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