The characterisation and identification of glycosyltransferases (GTs) involved in matrix polysaccharide biosynthesis has gained increasing interest as they affect our ability to influence, engineer and manipulate plant cell wall biosynthesis. To date, only a small number of genes coding for matrix polysaccharide biosynthesis-related glycosyltransferases have been identified. Mostly, biochemical proof of activity is still needed for final confirmation of function. To address such problems, this study established an initial biochemical activity testing methodology combined with final gene identification in one approach. A substrate library was established with different sugar nucleotide donor and oligosaccharide acceptor combinations to address a comprehensive range of glycosyltransferase activities. To overcome well-known problems with radioactive assay formats, an APTS (8-Amino- 1,3,6,PyreneTriSulfonic acid) based detection methodology was introduced, which allowed sensitive and flexible analysis of the glycosyltransferases.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:580570 |
| Date | January 2012 |
| Creators | Götz, Stephan |
| Publisher | University of East Anglia |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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