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dc.contributor.advisorBall, Judth M
dc.creatorAguirre, Rene 1989-
dc.date.accessioned2013-06-04T16:11:48Z
dc.date.available2013-06-04T16:11:48Z
dc.date.created2011-05
dc.date.issued2011-05-04
dc.date.submittedMay 2011
dc.identifier.urihttp://hdl.handle.net/1969.1/ETD-TAMU-2011-05-9680
dc.identifier.urihttp://hdl.handle.net/1969.1/148801
dc.description.abstractRotavirus (RV) is a viral pathogen that infects everyone worldwide. It initially infects the enterocytes of the small intestine causing severe diarrhea in affected patients, primarily the young. RV nonstructural protein 4 (NSP4) was the first identified enterotoxin associated with RV infections. Our main focus was to understand how this protein interacted with infected host-cell molecules, thus inducing disease symptoms. We utilized a yeast expression system that had been transformed with a plasmid to express a truncated form of NSP4 (tNSP4) which still possessed toxic characteristics. Before we could study key interactions of the enterotoxin, isolation and purification of NSP4 was required. Purification was done by the use of gel filtration (GF) chromatography, and the use of an affinity column to complete the final purification steps. Our samples were analyzed for purity by protein quantitation, SDS-PAGE, Silver Stain, and Western Blot. Once purified, the tNSP4 was to be chemically linked to magnetic beads, which were then to be incubated with RV infected cell lysates to "capture‟ interacting host-cell molecules. The bound cellular proteins will be excised from 2-D gels and submitted to the TAMU Mass Spectroscopy Core for sequencing. A BLAST search of the recovered sequences against available databases will identify the interacting host-cell proteins.
dc.format.mimetypeapplication/pdf
dc.subjectPurufication of NSP4
dc.subjectEnterotoxin
dc.subjectNSP4
dc.subjectNonstructural Protein 4
dc.subjectRotavirus
dc.titlePURIFICATION OF THE ROTAVIRUS NSP4 ENTEROTOXIN AND DISCERNMENT OF KEY HOST-CELL INTERACTIONS THAT CONTRIBUTE TO NSP4 SECRETION FROM THE CELL
dc.typeThesis
thesis.degree.departmentVeterinary Pathobiology
thesis.degree.disciplineBiomedical Sciences
thesis.degree.grantorHonors and Undergraduate Research
thesis.degree.nameBachelor of Science
dc.type.materialtext
dc.date.updated2013-06-04T16:11:48Z


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