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Access Type
WSU Access
Date of Award
January 2024
Degree Type
Thesis
Degree Name
M.S.
Department
Biochemistry and Molecular Biology
First Advisor
Jeffrey H. Withey
Abstract
Vibrio cholerae is a gram-negative, aquatic bacterium responsible for the deadly diarrheal disease cholera, which affects millions worldwide each year. The zebrafish (Danio rerio), indigenous to cholera endemic regions, serves as an ideal model organism due to its natural susceptibility to V. cholerae infection. However, the specific genetic factors important for Vibrio cholerae colonization in zebrafish are unknown. Previous studies in the lab utilized transposon sequencing analysis (Tn-Seq) to identify potential colonization factors required by V. cholerae in the zebrafish intestine. Some genes identified in this manner were observed to be associated with transport and transcriptional regulation. A few selected genes were subjected to a complete gene knockout (KO) in the El Tor C6706 wild type (WT) strain. The KO strains were generated using Splicing by Overlap Extension method, followed by chitin-induced natural transformation. Using zebrafish as a model, parallel infections were conducted between the KO and WT strains to determine if the suspected colonization factor is important for colonization. We hypothesize that the mutants created will have significant changes in colonizing the zebrafish intestine compared to the wild type. However, our colonization experiments with Δ TCS sensor histidine kinase and Δ Cation-proton antiporter revealed that the gene deletions have no significant impact on intestinal colonization. Identifying genes important for fish colonization holds immense potential and will contribute to our broader understanding of bacterial pathogenesis.
Recommended Citation
Rao, Apoorva, "Characterization Of Vibrio Cholerae Colonization Factors In Zebrafish" (2024). Wayne State University Theses. 955.
https://digitalcommons.wayne.edu/oa_theses/955