Full Text Available
Note: Clicking the button above will open the full text document at the original institutional repository in a new window.
Biopolymer gene discovery and characterization using metagenomic libraries
Thesis (MSc (Genetics. Institute of Plant Biotechnology))--Stellenbosch University, 2008.
Saved in:
| Main Author: | |
|---|---|
| Other Authors: | |
| Format: | Thesis |
| Language: | English |
| Published: |
Stellenbosch : Stellenbosch University
2008
|
| Subjects: | |
| Tags: |
No Tags, Be the first to tag this record!
|
| _version_ | 1867613794824355840 |
|---|---|
| access_status_str | Open Access |
| author | Ohlhoff, Colin Walter |
| author2 | Lloyd, James Richard |
| author_browse | Lloyd, James Richard Ohlhoff, Colin Walter |
| author_facet | Lloyd, James Richard Ohlhoff, Colin Walter |
| author_sort | Ohlhoff, Colin Walter |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis (MSc (Genetics. Institute of Plant Biotechnology))--Stellenbosch University, 2008. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/2380 |
| institution | Stellenbosch University (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:41:46.856Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2008 |
| publishDateRange | 2008 |
| publishDateSort | 2008 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/2380 Biopolymer gene discovery and characterization using metagenomic libraries Ohlhoff, Colin Walter Lloyd, James Richard Bauer, R. Kossmann, J. M. Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics. Institute for Plant Biotechnology. Metagenomics Metagenomic libraries Beta glucans Functional screening Biopolymer gene discovery Theses -- Plant biotechnology Dissertations -- Plant biotechnology Biopolymers Genetics Institute for Plant Biotechnology Thesis (MSc (Genetics. Institute of Plant Biotechnology))--Stellenbosch University, 2008. Traditional methods used for the discovery of novel genes have previously relied upon the ability to culture the relevant microbes and then demonstrate the activity of a specific enzyme. Although these methods have proved successful in the past, they severely limit our access to the genomes of organisms which are not able to be cultured under laboratory conditions. It was therefore the aim of this project to use metagenomic strategies for the identification of novel polymer-producing genes with the prospect of commercial exploitation. In this study, soil-derived metagenomic libraries were functionally screened for potential -glucan producing clones using aniline blue staining. Positive reacting clones were selected and sequenced. Initial sequencing revealed a gene with high homology to previously described glucan synthases, the products of these genes all having significant industrial value. The clone was transformed into a suitable bacterial host, cultured and allowed to produce the polymer of interest. The polysaccharide was purified and subjected to various chemical analyses so as to confirm its monosaccharide composition. Data suggests that this polymer is composed mainly of glucose units and that it may be secreted out of the cell. Purification of the active enzyme was attempted using classical protein purification methods with faint activity being detected using Native polyacrylamide gel electrophoresis (PAGE). Further attempts to demonstrate activity were made through the construction of a GST (glutathione S-transferase) tagged fusion protein. The second part of this study focuses on the construction and screening of a metagenomic DNA library from whey, a by-product of the cheese manufacturing process. It was envisaged that this could provide a resource for the identification of high value polymers when lactose is provided as a sole carbon source. The library was screened for function using Congo Red for the detection of extra-cellular polysaccharides. Masters 2008-11-19T12:13:26Z 2010-06-01T08:47:29Z 2008-11-19T12:13:26Z 2010-06-01T08:47:29Z 2008-12 Thesis http://hdl.handle.net/10019.1/2380 en Stellenbosch University application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Metagenomics Metagenomic libraries Beta glucans Functional screening Biopolymer gene discovery Theses -- Plant biotechnology Dissertations -- Plant biotechnology Biopolymers Genetics Institute for Plant Biotechnology Ohlhoff, Colin Walter Biopolymer gene discovery and characterization using metagenomic libraries |
| title | Biopolymer gene discovery and characterization using metagenomic libraries |
| title_full | Biopolymer gene discovery and characterization using metagenomic libraries |
| title_fullStr | Biopolymer gene discovery and characterization using metagenomic libraries |
| title_full_unstemmed | Biopolymer gene discovery and characterization using metagenomic libraries |
| title_short | Biopolymer gene discovery and characterization using metagenomic libraries |
| title_sort | biopolymer gene discovery and characterization using metagenomic libraries |
| topic | Metagenomics Metagenomic libraries Beta glucans Functional screening Biopolymer gene discovery Theses -- Plant biotechnology Dissertations -- Plant biotechnology Biopolymers Genetics Institute for Plant Biotechnology |
| url | http://hdl.handle.net/10019.1/2380 |
| work_keys_str_mv | AT ohlhoffcolinwalter biopolymergenediscoveryandcharacterizationusingmetagenomiclibraries |