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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

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.

Identiferoai:union.ndltd.org:BRADFORD/oai:bradscholars.brad.ac.uk:10454/5344
Date January 2011
CreatorsAl-Johani, Nasser D.
ContributorsPicksley, Stephen M., Elbashbehs, H., Fletcher, Jonathan N.
PublisherUniversity of Bradford, School of Life Sciences
Source SetsBradford Scholars
LanguageEnglish
Detected LanguageEnglish
TypeThesis, research masters, MPhil
Rights<a rel="license" href="http://creativecommons.org/licenses/by-nc-nd/3.0/"><img alt="Creative Commons License" style="border-width:0" src="http://i.creativecommons.org/l/by-nc-nd/3.0/88x31.png" /></a><br />The University of Bradford theses are licenced under a <a rel="license" href="http://creativecommons.org/licenses/by-nc-nd/3.0/">Creative Commons Licence</a>.

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