Investigation of Streptomyces promoters

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dc.contributor.advisorThomson, Jennifer Annen_ZA
dc.contributor.advisorKirby, Ralphen_ZA
dc.contributor.authorBourn, William Richarden_ZA
dc.date.accessioned2014-11-15T19:38:33Z
dc.date.available2014-11-15T19:38:33Z
dc.date.issued1995en_ZA
dc.descriptionBibliography: leaves 221-[234].en_ZA
dc.description.abstract[The work described here had multiple aims: to create a promoter probe that was suitable for the isolation of developmentally regulated Strepcomyces promoters, to isolate such promoters, to develop a computer assisted analysis system whereby potential promoter sequences could be determined and to use this in the analysis of the cloned promoters. Initially the suitability of the Streptomyces antibioticus me1C operon for use as a reporter system in Streptomyces was investigated. It was established that late-expressed promoters could be identified and that it was possible to use the me1C2 gene alone for this purpose. However, it was shown that the use of both me1C1 and me1C2 resulted in a more sensitive reporter system. High copy number promoter probe vectors were constructed and tested. A low copy number promoter probe (which used the Streptomyces penemefaciens pSPN1 origin of replication) was also constructed. The characteristics (copy number, stability and mobility) of the probe were established. The conditions in which sporulation was induced by phosphate limitation were identified. Under such conditions late expressing, phosphate dependent promoters were isolated, using the promoter probes previously developed. The expression of these promoters was tested in Streptomyces coelicolor bldA mutants, and the bldA dependent promoters identified. These were sequenced. Computer assisted analysis of DNA sequence bias was conducted, with the intention of using bias patterns to identify potential regulatory regions. The initial approach of using the sequence bias of protein coding regions (based on the premise that regulatory sites are likely to be under represented in these regions) was unsuccessful. Further analysis in which the positional preference of sequences that were over represented in regulatory regions was conducted. Based on this the known promoters of Streptomyces were partially classified. The sequence bias of protein coding DNA regions was used to develop a novel method to identify the protein coding regions of Streptomyces DNA. The computer programs were then used to identify protein coding and potential regulatory regions.en_ZA
dc.identifier.apacitationBourn, W. R. (1995). <i>Investigation of Streptomyces promoters</i>. (Thesis). University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology. Retrieved from http://hdl.handle.net/11427/9651en_ZA
dc.identifier.chicagocitationBourn, William Richard. <i>"Investigation of Streptomyces promoters."</i> Thesis., University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 1995. http://hdl.handle.net/11427/9651en_ZA
dc.identifier.citationBourn, W. 1995. Investigation of Streptomyces promoters. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Bourn, William Richard AB - [The work described here had multiple aims: to create a promoter probe that was suitable for the isolation of developmentally regulated Strepcomyces promoters, to isolate such promoters, to develop a computer assisted analysis system whereby potential promoter sequences could be determined and to use this in the analysis of the cloned promoters. Initially the suitability of the Streptomyces antibioticus me1C operon for use as a reporter system in Streptomyces was investigated. It was established that late-expressed promoters could be identified and that it was possible to use the me1C2 gene alone for this purpose. However, it was shown that the use of both me1C1 and me1C2 resulted in a more sensitive reporter system. High copy number promoter probe vectors were constructed and tested. A low copy number promoter probe (which used the Streptomyces penemefaciens pSPN1 origin of replication) was also constructed. The characteristics (copy number, stability and mobility) of the probe were established. The conditions in which sporulation was induced by phosphate limitation were identified. Under such conditions late expressing, phosphate dependent promoters were isolated, using the promoter probes previously developed. The expression of these promoters was tested in Streptomyces coelicolor bldA mutants, and the bldA dependent promoters identified. These were sequenced. Computer assisted analysis of DNA sequence bias was conducted, with the intention of using bias patterns to identify potential regulatory regions. The initial approach of using the sequence bias of protein coding regions (based on the premise that regulatory sites are likely to be under represented in these regions) was unsuccessful. Further analysis in which the positional preference of sequences that were over represented in regulatory regions was conducted. Based on this the known promoters of Streptomyces were partially classified. The sequence bias of protein coding DNA regions was used to develop a novel method to identify the protein coding regions of Streptomyces DNA. The computer programs were then used to identify protein coding and potential regulatory regions. DA - 1995 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 1995 T1 - Investigation of Streptomyces promoters TI - Investigation of Streptomyces promoters UR - http://hdl.handle.net/11427/9651 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/9651
dc.identifier.vancouvercitationBourn WR. Investigation of Streptomyces promoters. [Thesis]. University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 1995 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/9651en_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.otherMicrobiologyen_ZA
dc.titleInvestigation of Streptomyces promotersen_ZA
dc.typeDoctoral Thesis
dc.type.qualificationlevelDoctoral
dc.type.qualificationnamePhDen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
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