Attempt to express the Xerophyta viscose stress-responsive gene, Xvcor, in yeast with view to functional analysis

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dc.contributor.authorTawouokam, Jean Bernarden_ZA
dc.date.accessioned2014-07-30T17:42:04Z
dc.date.available2014-07-30T17:42:04Z
dc.date.issued2002en_ZA
dc.descriptionBibliography: leaves 24-29.
dc.description.abstractLow temperature is one of the environmental factors that cause substantial crop losses in the world. Recent advances in the study of plants native from temperate regions, have established that cold acclimation is regulated at the gene expression leveL To study the function ofaXerophyta viscosa cold responsive protein that accumulates in plant cells under various environmental stresses, we have cloned the Xerophyta viscosa cold-responsive cDNA gene designed Xvcor, into a less complex host, Saccharomyces cerevisiae with the purpose of carrying out in vivo functional analysis. Sequence analysis showed that Xvcor encodes a 264 amino acid residue protein (Garwe et al 2002). The Hydropathy plot indicated that the XVCOR protein is highly hydrophobic and contains 6 transmembrane domains (Garwe et al 2002). In order to achieve high-level expression of Xvcor, the gene was placed under the control of a strong promoter (Phosphoglycerate kinase). Reverse-transcription PCR amplification revealed that the Xvcor transcript accumulated in yeast cells. However, SDS-PAGE analysis could not detect the predicted 29.6 kDa recombinant protein, suggesting that the translation might be hampered. Tests designed to measure the enhancement of stress tolerance between the recombinant and the control showed similar growth performance, confirming the absence of recombinant protein. A computer search for codon usage showed that the codon usage bias in Xvcor was low compared with that of highly expressed genes of S. cerevisiae. Together, these results suggest that the codon usage in Xvcor could influence its expression in yeast.en_ZA
dc.identifier.apacitationTawouokam, J. B. (2002). <i>Attempt to express the Xerophyta viscose stress-responsive gene, Xvcor, in yeast with view to functional analysis</i>. (Thesis). University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology. Retrieved from http://hdl.handle.net/11427/4338en_ZA
dc.identifier.chicagocitationTawouokam, Jean Bernard. <i>"Attempt to express the Xerophyta viscose stress-responsive gene, Xvcor, in yeast with view to functional analysis."</i> Thesis., University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 2002. http://hdl.handle.net/11427/4338en_ZA
dc.identifier.citationTawouokam, J. 2002. Attempt to express the Xerophyta viscose stress-responsive gene, Xvcor, in yeast with view to functional analysis. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Tawouokam, Jean Bernard AB - Low temperature is one of the environmental factors that cause substantial crop losses in the world. Recent advances in the study of plants native from temperate regions, have established that cold acclimation is regulated at the gene expression leveL To study the function ofaXerophyta viscosa cold responsive protein that accumulates in plant cells under various environmental stresses, we have cloned the Xerophyta viscosa cold-responsive cDNA gene designed Xvcor, into a less complex host, Saccharomyces cerevisiae with the purpose of carrying out in vivo functional analysis. Sequence analysis showed that Xvcor encodes a 264 amino acid residue protein (Garwe et al 2002). The Hydropathy plot indicated that the XVCOR protein is highly hydrophobic and contains 6 transmembrane domains (Garwe et al 2002). In order to achieve high-level expression of Xvcor, the gene was placed under the control of a strong promoter (Phosphoglycerate kinase). Reverse-transcription PCR amplification revealed that the Xvcor transcript accumulated in yeast cells. However, SDS-PAGE analysis could not detect the predicted 29.6 kDa recombinant protein, suggesting that the translation might be hampered. Tests designed to measure the enhancement of stress tolerance between the recombinant and the control showed similar growth performance, confirming the absence of recombinant protein. A computer search for codon usage showed that the codon usage bias in Xvcor was low compared with that of highly expressed genes of S. cerevisiae. Together, these results suggest that the codon usage in Xvcor could influence its expression in yeast. DA - 2002 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 2002 T1 - Attempt to express the Xerophyta viscose stress-responsive gene, Xvcor, in yeast with view to functional analysis TI - Attempt to express the Xerophyta viscose stress-responsive gene, Xvcor, in yeast with view to functional analysis UR - http://hdl.handle.net/11427/4338 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/4338
dc.identifier.vancouvercitationTawouokam JB. Attempt to express the Xerophyta viscose stress-responsive gene, Xvcor, in yeast with view to functional analysis. [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/11427/4338en_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 cell biologyen_ZA
dc.titleAttempt to express the Xerophyta viscose stress-responsive gene, Xvcor, in yeast with view to functional analysisen_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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