dc.contributor.author | Thomas, Stanley | |
dc.date.accessioned | 2014-04-15T18:37:37Z | |
dc.date.available | 2014-04-15T18:37:37Z | |
dc.date.issued | 2006-07-01 | |
dc.identifier.uri | https://hdl.handle.net/11244/8502 | |
dc.description.abstract | The objectives of this study were to over-express the sheep prion protein (RecShPrPC) using a bacterial vector. The prion protein gene was PCR amplified using sheep genomic DNA and was cloned into pET102/D-TOPO plasmid. The integrity of the cloned PCR product as well as the His-tagged fusion protein and its orientation within the vector was determined by DNA sequencing and upon confirmation was induced with isopropyl-beta-D-thiogalactopyranoside for protein expression. The expressed proteins were solubilized by addition of 6M guanidinium chloride in lysis buffer and purified using adsorption to a Nickel-Nitriloacetic++ metal affinity resin. Production of RecShPrPC protein by this method could be used to study of solubilzation and fractionation of prion proteins from muscle proteins. Separation of prion proteins from the myofibrillar and sarcoplasmic proteins may enhance the safety of meat products that would eventually be rendered and used for non-mammalian animal feed or fertilizer. | |
dc.format | application/pdf | |
dc.language | en_US | |
dc.publisher | Oklahoma State University | |
dc.rights | Copyright is held by the author who has granted the Oklahoma State University Library the non-exclusive right to share this material in its institutional repository. Contact Digital Library Services at lib-dls@okstate.edu or 405-744-9161 for the permission policy on the use, reproduction or distribution of this material. | |
dc.title | Production of Recombinant Sheep Prion Protein (Recshprpc) and Its Detection in Muscle Food Using Western Blotting | |
dc.type | text | |
osu.filename | Thomas_okstate_0664M_1959.pdf | |
osu.college | Agricultural Sciences and Natural Resources | |
osu.accesstype | Open Access | |
dc.description.department | Department of Animal Science | |
dc.type.genre | Thesis | |