Honors Theses

Date of Award

Spring 5-22-2020

Document Type

Undergraduate Thesis

Department

Biology

First Advisor

Bradley Jones

Second Advisor

Josh Bloomekatz

Third Advisor

Gregg Roman

Relational Format

Dissertation/Thesis

Abstract

Apoptosis is the process of programmed cell death. Increasing knowledge of this biological process can lead to understanding of neurodegenerative diseases and cancer alike. In mammals, this process occurs through mechanisms that are localized to the mitochondria. The release of cytochrome c is from the mitochondrial outer membrane (MOM) is regulated by pro and anti-apoptotic Bcl-2 family proteins. If the latter are successful, then cytochrome c’s release into the cytosol activates a caspase cascade, resulting in apoptosis of the cell. Pro-apoptotic Bax is present in complexes that contain voltage dependent anion channels (VDACs). We hypothesized that in healthy cells, VDACs act as a restraint by creating a complex in the MOM that interferes with Bax’s ability to form pores with Bak.

Drosophila Melanogaster was used as the genetic platform to create and induce mutations in the porin gene. The UAS/GAL4 system is used to target gene expression within any tissue of the Drosophila organism. We used transgenic flies using this system to enhance the expression of Bax in the eye or wing tissues of the fly. When Bax is expressed it induces apoptosis. We also used the CRISPR/Cas9 system to introduce point mutations within the BH3 domain of the Drosophila form of VDAC (Porin) to investigate the Bax-Porin relationship.

We then used polymerase chain reaction (PCR) and gel electrophoresis to determine the presence of the mutation.

Accessibility Status

Searchable text

Included in

Biology Commons

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.