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dc.contributor.advisorLangeland, James A., 1964-
dc.contributor.advisorLacy, D. Borden
dc.contributor.authorThomas, Audrey
dc.descriptionvi, 70 p.en_US
dc.description.abstractClostridium difficile infections in the colon cause severe diarrhea and pseudomembranous colitis, with pathology stemming from two secreted bacterial toxins, TcdA and TcdB. With C. difficile infection rates on the rise in the United States, and with the growing threat of antibiotic resistant strains, hospitals are desperate for more effective treatments. Previous research has shown that the production of TcdA and TcdB depends on a regulatory sigma factor, TcdR, which directs RNA polymerase to the promoter regions of the toxins in order to initiate transcription. If TcdR function were to be knocked out by a small-molecule therapeutic, toxin production and subsequent C. difficile virulence could be reduced. Determining the protein structure of TcdR, as well as understanding its functionality in relation to RNA polymerase and toxin promoter regions, could accelerate inhibitor development. In this study, TcdR was expressed in Escherichia coli, and purified with co-eluting proteins by affinity and size-exclusion chromatography. Using mass spectrometry and electron microscopy, we were able to characterize a putative complex between TcdR and E. coli RNA polymerase. Electrophoretic mobility shift assays are currently being optimized to confirm the ability of the putative complex to bind toxin promoter DNA. The continuation of these studies will provide the basis for future functional screens for small-molecule inhibitors against the TcdR/RNA polymerase complex that could help combat C. difficile virulence and infection.en_US
dc.publisherKalamazoo Collegeen_US
dc.relation.ispartofKalamazoo College Biology Senior Individualized Projects Collection
dc.rightsU.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.
dc.titleStructural Characterization of Clostridium difficile TcdR, a Sigma Factor that Controls Bacterial Toxin Productionen_US
KCollege.Access.ContactIf you are not a current Kalamazoo College student, faculty, or staff member, email to request access to this thesis.

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  • Biology Senior Individualized Projects [1549]
    This collection includes Senior Individualized Projects (SIP's) completed in the Biology Department. Abstracts are generally available to the public, but PDF files are available only to current Kalamazoo College students, faculty, and staff.

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