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Identification of South African fungi with high protein secreting capabilities
Dissertation (MSc)--University of Pretoria, 2015.
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| Other Authors: | |
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| Format: | Thesis |
| Language: | English |
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University of Pretoria
2016
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| _version_ | 1867613500894871552 |
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| access_status_str | Open Access |
| author2 | Cowan, Don A. |
| author_browse | Cowan, Don A. |
| author_facet | Cowan, Don A. |
| collection | Thesis |
| dc_rights_str_mv | © 2016, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc)--University of Pretoria, 2015. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/53503 |
| institution | University of Pretoria (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:37:08.629Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2016 |
| publishDateRange | 2016 |
| publishDateSort | 2016 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/53503 Identification of South African fungi with high protein secreting capabilities Cowan, Don A. u13321049@tuks.co.za Berger, E. Kabwe, Mubanga Helen UCTD Dissertation (MSc)--University of Pretoria, 2015. Recombinant protein technology has facilitated high level production of proteins to try and meet the demand from industry. One major factor that has been reported to influence the scales of production in industrial settings is the relationship between host cell growth rate and protein production rate. Therefore, expression systems with fast growth and high protein production rates are favorable for high volume recombinant protein production. Filamentous fungi have over the years been used to produce a range of valuable metabolites including biologically active proteins and organic acids. Their ability to synthesize and secrete extracellular enzymes in large amounts made them the preferred host systems for heterologous protein production. In this study the CMW: Culture Collection of the Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, South Africa, was screened using SDS-PAGE gels to identify a fungal isolate with the natural ability to secrete a highly expressed protein (HEP). The HEP was identified through liquid chromatography-mass spectrometry (LC-MS/MS) and the 330 bp gene encoding the HEP was isolated, cloned and transformed into E. coli cells. The gene sequence was further investigated using a modified SiteFinding-PCR chromosome walking technique, to acquire the full man gene sequence and to identify the upstream promoter and regulatory sequences flanking the gene in the 5 to 3 direction. In silico analysis of acquired nucleotide sequences in the 5 direction promoter region of the gene, predicted transcription initiation sites, transcription factor binding sites as well as TATA boxes that have been previously identified in the promoter region of other Ophiostoma proteins. The growth conditions of O. phasma 20676 were also optimized for high level protein production. Microbiology and Plant Pathology MSc Unrestricted 2016-07-01T10:33:08Z 2016-07-01T10:33:08Z 2016-04-18 2015 Dissertation Kabwe, MH 2016, Identification of South African fungi with high protein secreting capabilities, MSc Dissertation, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/53503> A2016 http://hdl.handle.net/2263/53503 en © 2016, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | UCTD Identification of South African fungi with high protein secreting capabilities |
| title | Identification of South African fungi with high protein secreting capabilities |
| title_full | Identification of South African fungi with high protein secreting capabilities |
| title_fullStr | Identification of South African fungi with high protein secreting capabilities |
| title_full_unstemmed | Identification of South African fungi with high protein secreting capabilities |
| title_short | Identification of South African fungi with high protein secreting capabilities |
| title_sort | identification of south african fungi with high protein secreting capabilities |
| topic | UCTD |
| url | http://hdl.handle.net/2263/53503 |