Investigating ybeX/corC Mutation and Phenotypic Changes in Escherichia coli and Salmonella typhimurium

Patel, Tanvi

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Year:: 2023
Contributor:: Patel, Tanvi (creator); Nau, Gerard (Advisor); Schell, Jacquelyn (Reader); Fawzi, Nicolas (Reader); Brown University. Department of Molecular Pharmacology, Physiology and Biotechnology (sponsor)
Genre:: theses
Subject:: Genomics; antimicrobial resistance
Extent:: , None p.

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Abstract:: Antimicrobial resistance is a major problem and according to Centers for Disease Control and Prevention (CDC) it causes about 2.8 million antimicrobial-resistant infections every year in the U.S. Hence, identifying newer antimicrobial targets is essential. Genomic studies are useful in identifying genes that contribute to the pathogenicity of microbial infections. Often drugs are designed to target genes that are conserved across various bacterial species but absent in humans. ybeX, and its homologous gene corC, is found in various bacteria such as Escherichia coli, Salmonella typhi, Shigella flexneri, Klebsiella pneumoniae, and Pseudomonas aeruginosa. In this research, ybeX/corC gene was studied in E. coli BW25113 and S. typhimurium LT2 strains. A previous Nau Laboratory student mutated ybeX in E. coli using a two-step homologous recombination strategy which inserted the tetA-sacB cassette, followed by an ybeX allele with a stop codon (ybeXstop) into the ybeX locus of the wild-type E. coli strain. ybeX mutation in E. coli was verified via techniques like allele-specific PCR and Sanger sequencing. Parallel studies of antibiotic susceptibility testing were conducted in E. coli. In addition to that, a substitution mutation in corC of S. typhimurium was also studied in this work and an attempt to complement the corC mutation in S. typhimurium using a plasmid with an arabinose-inducible promoter 5’ to the wild-type E. coli ybeX was made. Swimming motility phenotype was tested in wild-type, mutated and genetically complemented strains of S. typhimurium. Results indicated that tetA-sacB was not successfully replaced by the ybeXstop allele, instead a more complex recombination event occurred at the ybeX locus. It was also revealed that although the E. coli ybeX and S. typhimurium corC are 97.3% similar, complementing the corC mutation in S. typhimurium strain with E. coli ybeX does not return the wild-type swimming motility phenotype.
Notes:: Thesis (Sc. M.)--Brown University, 2023

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Citation

Patel, Tanvi, "Investigating ybeX/corC Mutation and Phenotypic Changes in Escherichia coli and Salmonella typhimurium" (2023). Molecular Pharmacology, Physiology, and Biotechnology Theses and Dissertations. Brown Digital Repository. Brown University Library. https://repository.library.brown.edu/studio/item/bdr:7k7kvavq/

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Molecular Pharmacology, Physiology, and Biotechnology Theses and Dissertations

Theses and Dissertations for the Molecular Pharmacology, Physiology, and Biotechnology department.
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