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An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors
Bibliography: pages 80-90.
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| Main Author: | |
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| Other Authors: | |
| Format: | Thesis |
| Language: | English |
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Department of Molecular and Cell Biology
2016
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| _version_ | 1867613230217560064 |
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| access_status_str | Open Access |
| author | Gehringer, Michelle Martine |
| author2 | Rybicki, Edward P |
| author_browse | Gehringer, Michelle Martine Rybicki, Edward P |
| author_facet | Rybicki, Edward P Gehringer, Michelle Martine |
| author_sort | Gehringer, Michelle Martine |
| collection | Thesis |
| description | Bibliography: pages 80-90. |
| format | Thesis |
| id | oai:open.uct.ac.za:11427/19709 |
| institution | University of Cape Town (South Africa) |
| language | eng |
| last_indexed | 2026-06-10T12:32:50.328Z |
| license_str | Not specified — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository |
| publishDate | 2016 |
| publishDateRange | 2016 |
| publishDateSort | 2016 |
| publisher | Department of Molecular and Cell Biology |
| publisherStr | Department of Molecular and Cell Biology |
| record_format | dspace |
| source_str | UCTD — University of Cape Town Open Access Repository |
| spelling | oai:open.uct.ac.za:11427/19709 An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors Gehringer, Michelle Martine Rybicki, Edward P Microbiology Bibliography: pages 80-90. 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. 2016-05-18T07:13:16Z 2016-05-18T07:13:16Z 1996 Master Thesis Masters MSc http://hdl.handle.net/11427/19709 eng application/pdf Department of Molecular and Cell Biology Faculty of Science University of Cape Town |
| spellingShingle | Microbiology Gehringer, Michelle Martine An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors |
| thesis_degree_str | Master's |
| title | An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors |
| title_full | An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors |
| title_fullStr | An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors |
| title_full_unstemmed | An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors |
| title_short | An investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors |
| title_sort | investigation into the high level production of proteins in tobacco using transgenic plants or viral vectors |
| topic | Microbiology |
| url | http://hdl.handle.net/11427/19709 |
| work_keys_str_mv | AT gehringermichellemartine aninvestigationintothehighlevelproductionofproteinsintobaccousingtransgenicplantsorviralvectors AT gehringermichellemartine investigationintothehighlevelproductionofproteinsintobaccousingtransgenicplantsorviralvectors |