The isolation and characterization of heat shock protein Hsp12 in Lipomyces starkeyi

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dc.contributor.advisorBrandt, Wolf Fen_ZA
dc.contributor.advisorLindsey, George Gen_ZA
dc.contributor.authorMukwevho, Emmanuelen_ZA
dc.date.accessioned2015-01-02T09:04:17Z
dc.date.available2015-01-02T09:04:17Z
dc.date.issued2002en_ZA
dc.descriptionBibliography: leaves 60-72.en_ZA
dc.description.abstractThe stress response protein Hsp 12 is induced in S. cerevisiae cells upon exposure to salt stress, heat shock, ethanol, and upon entry to stationary phase (Mtwisha et aI., 1998). In this study, the occurrence of proteins related to Hsp12 was investigated in a number of yeasts (namely, Saccharomyces cerevisiae S288C, Schizosaccharomyces pombe, Debaromyces hansenii, Lipomyces starkeyi Y-2024, Saccharomyces cerevisiae IFO 23X7 (Kaokai), Zygosaccharomyces rouxii and Pichia sorbitophila. This was performed by selective protein extraction followed by SDS-P AGE and western blotting using a S. cerevisiae anti-Hsp 12 antibody. The results showed that almost all the yeasts investigated possessed a protein that had an identical migration to that of Hsp 12 with the exception of S. pombe, which contained a 9 kDa protein. Western blotting using the antiHsp 12 antibody cross-reacted only with the two S. cerevisiae species in addition to the 12 kDa protein from Lipomyces starkeyi of all the species investigated. MALDI-TOF peptide mass analysis after tryptic digestion of the L. starkeyi 12 kDa protein showed that a close sequence similarity existed to that of S. cerevisiae Hsp 12 and none to rest of the 12 kDa proteins isolated from all the other species investigated. In order to determine the sequence of the Hsp 12 protein, the L. starkeyi Hsp 12 gene was amplified using S. cerevisiae Hsp 12 primers. Gene sequencing of both S. cerevisiae and L. starkeyi Hsp 12 genes revealed three nucleotide differences existed between them. L. starkeyi Hsp 12 was found to be present in relatively small amounts during early growth stages but increased during log phase with a slight further increase during stationary phase. Increasing the salt concentration in the growth medium was found to induce Hsp 12. Increased levels of Hsp 12 appeared to confer a degree of protection during desiccation and subsequent rehydration of both L. starkeyi and S. cerevisiae.en_ZA
dc.identifier.apacitationMukwevho, E. (2002). <i>The isolation and characterization of heat shock protein Hsp12 in Lipomyces starkeyi</i>. (Thesis). University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology. Retrieved from http://hdl.handle.net/11427/10953en_ZA
dc.identifier.chicagocitationMukwevho, Emmanuel. <i>"The isolation and characterization of heat shock protein Hsp12 in Lipomyces starkeyi."</i> Thesis., University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 2002. http://hdl.handle.net/11427/10953en_ZA
dc.identifier.citationMukwevho, E. 2002. The isolation and characterization of heat shock protein Hsp12 in Lipomyces starkeyi. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Mukwevho, Emmanuel AB - The stress response protein Hsp 12 is induced in S. cerevisiae cells upon exposure to salt stress, heat shock, ethanol, and upon entry to stationary phase (Mtwisha et aI., 1998). In this study, the occurrence of proteins related to Hsp12 was investigated in a number of yeasts (namely, Saccharomyces cerevisiae S288C, Schizosaccharomyces pombe, Debaromyces hansenii, Lipomyces starkeyi Y-2024, Saccharomyces cerevisiae IFO 23X7 (Kaokai), Zygosaccharomyces rouxii and Pichia sorbitophila. This was performed by selective protein extraction followed by SDS-P AGE and western blotting using a S. cerevisiae anti-Hsp 12 antibody. The results showed that almost all the yeasts investigated possessed a protein that had an identical migration to that of Hsp 12 with the exception of S. pombe, which contained a 9 kDa protein. Western blotting using the antiHsp 12 antibody cross-reacted only with the two S. cerevisiae species in addition to the 12 kDa protein from Lipomyces starkeyi of all the species investigated. MALDI-TOF peptide mass analysis after tryptic digestion of the L. starkeyi 12 kDa protein showed that a close sequence similarity existed to that of S. cerevisiae Hsp 12 and none to rest of the 12 kDa proteins isolated from all the other species investigated. In order to determine the sequence of the Hsp 12 protein, the L. starkeyi Hsp 12 gene was amplified using S. cerevisiae Hsp 12 primers. Gene sequencing of both S. cerevisiae and L. starkeyi Hsp 12 genes revealed three nucleotide differences existed between them. L. starkeyi Hsp 12 was found to be present in relatively small amounts during early growth stages but increased during log phase with a slight further increase during stationary phase. Increasing the salt concentration in the growth medium was found to induce Hsp 12. Increased levels of Hsp 12 appeared to confer a degree of protection during desiccation and subsequent rehydration of both L. starkeyi and S. cerevisiae. DA - 2002 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 2002 T1 - The isolation and characterization of heat shock protein Hsp12 in Lipomyces starkeyi TI - The isolation and characterization of heat shock protein Hsp12 in Lipomyces starkeyi UR - http://hdl.handle.net/11427/10953 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/10953
dc.identifier.vancouvercitationMukwevho E. The isolation and characterization of heat shock protein Hsp12 in Lipomyces starkeyi. [Thesis]. University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 2002 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/10953en_ZA
dc.language.isoengen_ZA
dc.publisher.departmentDepartment of Molecular and Cell Biologyen_ZA
dc.publisher.facultyFaculty of Scienceen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.subject.otherMolecular and Cellular Biologyen_ZA
dc.titleThe isolation and characterization of heat shock protein Hsp12 in Lipomyces starkeyien_ZA
dc.typeMaster Thesis
dc.type.qualificationlevelMasters
dc.type.qualificationnameMScen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
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