Essay
Functional analysis of the 152-156 amino acid region of yeast cytosine deaminase using alanine scanning mutagenesis
2004
Handle:
https://hdl.handle.net/2376/2415
Abstract
In order to accomplish this goal, the amino acids in residues 152-156 of the yCD genome were individually replaced with alanine. This substitution might allow greater binding of the yCD enzyme to 5FC. Five different mutants were generated in this process. They are W152A, F153A, E154A, D155A and Il56A. These mutants were tested using a plate assay for their ability to convert cytosine to uracil. This is the normal function of the yCD enzyme, so this test determined if the mutant yCD enzyme still had that ability. It was found that two of the mutant enzymes were able deaminate cytosine to create uracil, F153A and E154A. These mutants were further tested to see if they retained the ability to convert the prodrug 5FC to the toxic drug 5FU. It was determined that one mutant, E154A, had the ability to accomplish this conversion better than the normal (unmutated) enzyme. This signifies that is has great potential in suicide gene therapy and is undergoing further investigation.
Metrics
3 File views/ downloads
17 Record Views
Details
- Title
- Functional analysis of the 152-156 amino acid region of yeast cytosine deaminase using alanine scanning mutagenesis
- Creators
- Kathleen D. Bongiovanni (Author)
- Contributors
- Margaret E. Black (Advisor)
- Academic Unit
- Honors Theses (WSU Pullman, Passed with Distinction)
- Identifiers
- 99900590741301842
- Copyright
- http://rightsstatements.org/vocab/InC/1.0/; http://www.ndltd.org/standards/metadata; http://purl.org/eprint/accessRights/OpenAccess; In copyright; Publicly accessible; openAccess
- Language
- English
- Resource Type
- Essay