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Development of a Plasmid Containing the Bacillus anthracis cotE Gene

Development of a Plasmid Containing the Bacillus anthracis cotE Gene

dc.contributor.advisor Peterson, Nancy
dc.contributor.author Widel, Melissa
dc.contributor.editor Visick, Jonathon
dc.date.accessioned 2018-01-19T16:14:27Z
dc.date.available 2018-01-19T16:14:27Z
dc.date.issued 2017-05-15
dc.date.submitted 2017-05-15
dc.identifier.uri http://hdl.handle.net/10969/1211
dc.description.abstract Complex structures are formed and regulated by a network of gene and protein interactions. One such structure, the bacterial spore coat, is formed by sporulating members of the phyla Firmicutes, including species in the genus Bacillus. Formed in response to a harsh environment, a bacterial endospore is highly resistant due to this spore coat. The spore coat of Bacillus species can be used as a model system to study the role of genes in the formation and regulation of complex structures. In this study, development of a tool to study the morphogenic role of the Bacillus spore coat protein CotE began. Specifically, I worked to create a plasmid that would allow me to delete the cotE gene by homologous recombination resulting in an insertion event, thus allowing for the study of the role of CotE. A fragment of the cotE gene from Bacillus anthracis genomic DNA was amplified and ligated into an expression plasmid, which would allow for homologous recombination. Gel electrophoresis results indicate that the correct insert has been isolated, but successful insertion into the expression plasmid has yet yo be confirmed. Once the correct plasmid is confirmed, transformation and homologous recombination can result in an insertion deletion of cotE in B. anthracis. The relative conservation of cotE across species of the genus Bacillus should allow for the same engineered plasmid to be used to knock out cotE in multiple species of Bacillus. Successful completion of this research would present this method as a viable manner of researching complex structures. en_US
dc.language.iso en_US en_US
dc.title Development of a Plasmid Containing the Bacillus anthracis cotE Gene en_US
dc.type Thesis

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