An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors

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dc.contributor.advisorRybicki, Edward Pen_ZA
dc.contributor.authorGehringer, Michelle Martineen_ZA
dc.date.accessioned2016-05-18T07:13:16Z
dc.date.available2016-05-18T07:13:16Z
dc.date.issued1996en_ZA
dc.descriptionBibliography: pages 80-90.en_ZA
dc.description.abstractThe aim of this project was to construct a high level plant expression vector from the RNA 3 of cucumber mosaic virus strain Y (CMV Y). The 5'- and 3'-untranslated regions (UTRs) of this genome segment were reverse transcribed, cloned and sequenced. The chloramphenicol acetyl transferase gene (CAT) was inserted between the two UTRs. This artificial viral cDNA (5'cat3') was cloned immediately downstream of the cauliflower mosaic virus 35 S promoter at the transcription initiation site to make a DNA vector. An RNA vector construct was made by placing the 5'cat3' segment under the control of a T7 RNA promoter sequence. In vitro transcripts, as well as linearised DNA vector constructs were inoculated onto CMV infected plants. Inoculated plants were monitored for CAT expression. No CAT could be detected in total protein extracts of inoculated plants. No CAT mRNA could be detected in northern blots of total RNA extracted from inoculated plants. The vector constructed from the 5' - and 3' -UTR of the RNA 3 of CMV Y did not appear to contain all the necessary attributes for a viral expression vector. To study the expression of a foreign antigen in tobacco, the L 1 capsid protein of human papillomavirus type 16 was cloned into Agrobacterium tumefaciens and used to make transgenic Nicotiana tabacum. Kanamycin resistant tobacco plants were shown to carry the L 1 capsid gene using PCR screening, but western blots on total protein extracts of the transformed plants were indeterminate. Further studies are needed to determine whether the antigen is produced and if it is correctly processed.en_ZA
dc.identifier.apacitationGehringer, M. M. (1996). <i>An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors</i>. (Thesis). University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology. Retrieved from http://hdl.handle.net/11427/19709en_ZA
dc.identifier.chicagocitationGehringer, Michelle Martine. <i>"An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors."</i> Thesis., University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 1996. http://hdl.handle.net/11427/19709en_ZA
dc.identifier.citationGehringer, M. 1996. An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Gehringer, Michelle Martine AB - The aim of this project was to construct a high level plant expression vector from the RNA 3 of cucumber mosaic virus strain Y (CMV Y). The 5'- and 3'-untranslated regions (UTRs) of this genome segment were reverse transcribed, cloned and sequenced. The chloramphenicol acetyl transferase gene (CAT) was inserted between the two UTRs. This artificial viral cDNA (5'cat3') was cloned immediately downstream of the cauliflower mosaic virus 35 S promoter at the transcription initiation site to make a DNA vector. An RNA vector construct was made by placing the 5'cat3' segment under the control of a T7 RNA promoter sequence. In vitro transcripts, as well as linearised DNA vector constructs were inoculated onto CMV infected plants. Inoculated plants were monitored for CAT expression. No CAT could be detected in total protein extracts of inoculated plants. No CAT mRNA could be detected in northern blots of total RNA extracted from inoculated plants. The vector constructed from the 5' - and 3' -UTR of the RNA 3 of CMV Y did not appear to contain all the necessary attributes for a viral expression vector. To study the expression of a foreign antigen in tobacco, the L 1 capsid protein of human papillomavirus type 16 was cloned into Agrobacterium tumefaciens and used to make transgenic Nicotiana tabacum. Kanamycin resistant tobacco plants were shown to carry the L 1 capsid gene using PCR screening, but western blots on total protein extracts of the transformed plants were indeterminate. Further studies are needed to determine whether the antigen is produced and if it is correctly processed. DA - 1996 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 1996 T1 - An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors TI - An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors UR - http://hdl.handle.net/11427/19709 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/19709
dc.identifier.vancouvercitationGehringer MM. An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors. [Thesis]. University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 1996 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/19709en_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.titleAn investigation into the high level production of proteins in tobacco using transgenic plants or viral vectorsen_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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