CRISPR-Cas9 mediated xylose auxotrophy in Scheffersomyces stipitis

dc.contributor.advisor Robertson, James JBR
dc.contributor.author Johnson, Samuel Sheldon
dc.contributor.committeemember Elrod-Erickson, Matthew ME
dc.contributor.committeemember Nelson, David DN
dc.date.accessioned 2024-04-24T22:02:28Z
dc.date.available 2024-04-24T22:02:28Z
dc.date.issued 2024
dc.date.updated 2024-04-24T22:02:28Z
dc.description.abstract Scheffersomyces stipitis is a top xylose fermenter. The yeast efficiently ferments xylose into ethanol, making it industrially important. In this study, I (1) create a new toolset to knock out and replace any gene in the S. stipitis genome and (2) show the phenotypic alterations in the yeast's xylose fermentation after knocking out the HXK1 gene which encodes HXK1. This experiment compares sugar utilization in wild-type and HXK1 knockout strains. The unique toolset BLINCAR (Bioluminescent Indicator Nullified by Cas9 Actuated Recombination) demonstrated that I could knock out HXK1 in S. stipitis. I tested this utilizing molecular and yeast biotechnology. The results reveal that HXK1 knockouts utilize glucose similarly to the wild type. Since knocking out HXK1 does not cause xylose auxotrophy, other glucose phosphorylation enzymes might be responsible.
dc.description.degree M.S.
dc.identifier.uri https://jewlscholar.mtsu.edu/handle/mtsu/7181
dc.language.rfc3066 en
dc.publisher Middle Tennessee State University
dc.source.uri http://dissertations.umi.com/mtsu:11836
dc.subject Biology
dc.subject Bioengineering
dc.thesis.degreelevel masters
dc.title CRISPR-Cas9 mediated xylose auxotrophy in Scheffersomyces stipitis
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