Purification and Activity of the DnaK Heat Shock Protein of the Emerging Human Pathogen Rhodococcus equi. Optimisation of methods of purifying DnaK from Rhodococcus equi, and the use of the purified protein in assays to demonstrate its activity in isolation and with other heat shock proteins
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Publication date
2012-01-24Supervisor
Picksley, Stephen M.Elbashbehs, H.
Fletcher, Jonathan N.
Keyword
Heat shock proteins; DnaK
; Protein purification
; ATPase Assays
; Rhodococcus equi
; Opportunistic pathogen
Institution
University of BradfordDepartment
School of Life SciencesAwarded
2011Metadata
Show full item recordAbstract
Rhodococcus equi is an important pathogen in foals between one to six months of age and is a major cause of death in in these animals. In addition, R. equi has recently emerged as a significant opportunistic pathogen in immunosuppressed humans, especially those infected with HIV. Despite the ability of the organism to survive stressful growth conditions, for example, exposure to elevated temperature and oxygen radicals, the role of heat shock proteins in the pathogenesis of R. equi has not been well documented. In this project we developed and optimised methods to purify the heat shock protein DnaK from R. equi, using a combination of ion-exchange and affinity chromatography. The effectiveness of the purification protocols were assessed using SDS-PAGE and Western-blotting with anti-DnaK antibodies, and the enzymic activity of the purified DnaK was verified with an ATPase assay. ATPase assays were also used to investigate the roles of other heat shock proteins in enhancing the activity of DnaK.Type
ThesisQualification name
MPhilCollections
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