Heggie, Laura (1999) Approaching an RT-PCR Assay to Analyse Gene Expression in Chilling-Stressed Rhododendron: Partial Cloning of an Ascorbate Peroxidase Gene and Enzyme Activity Studies. PhD thesis, University of Glasgow.
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Abstract
Production and regulation of active oxygen species are important responses to environmental stress in plant tissues. This study was concerned with development of a competitive RT-PCR assay to study changes in ascorbate peroxidase gene expression in chilled and non-chilled in vitro grown cultures of Rhododendron ponticum, R. hatsugiri and R. impeditum. Oligonucleotides for PCR amplification of ascorbate peroxidase and glutathione reductase DNA sequences were designed using a sequence homology alignment of mRNA/DNA sequences from six distinct plant species. Ligation of PCR products into the pT-Adv plasmid vector and transformation into Escherichia coli, followed by partial sequencing, confirmed fragment identity. The subsequent design of Rhododendron- specific primers, and the construction of a cRNA competitor fragment by in vitro transcription for use in competitive RT-PCR, were also mediated by E. coli cloning. RT-PCR was developed using M-MLV reverse transcriptase and total RNA isolated from R. ponticum. The response of in vitro grown R. ponticum cultures upon exposure to chilling (4 and 2
Item Type: | Thesis (PhD) |
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Qualification Level: | Doctoral |
Additional Information: | Adviser: Rob Finch |
Keywords: | Genetics |
Date of Award: | 1999 |
Depositing User: | Enlighten Team |
Unique ID: | glathesis:1999-76252 |
Copyright: | Copyright of this thesis is held by the author. |
Date Deposited: | 19 Nov 2019 16:14 |
Last Modified: | 19 Nov 2019 16:14 |
URI: | https://theses.gla.ac.uk/id/eprint/76252 |
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