Development of stable isotope probing for the isolation of biocatalytic genes from aromatic compound-degrading bacteria
This thesis described the development of DNA Stable Isotope Probing (SIP) with the goal of identifying microorganisms and genes involved in the metabolism of aromatic compounds. The research had three main components. Results have been presented in chapters 3, 4 and 5. • Chapter 3 focused on investi...
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Queen's University Belfast
2012
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| Online Access: | http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.579722 |
| Summary: | This thesis described the development of DNA Stable Isotope Probing (SIP) with the goal of identifying microorganisms and genes involved in the metabolism of aromatic compounds. The research had three main components. Results have been presented in chapters 3, 4 and 5. • Chapter 3 focused on investigating the effect of growth on 2H labelled compounds, developing a method for the isolation of 2H labelled DNA and understanding the limitations of this approach using a range of pure cultures. • Chapter 4 describes experiments used to validate the 2H DNA SIP method using environmental samples and compare SIP results aiming to identify 16s rRNA and dioxygenase sequences using 2H and I3C labelled benzoic acid. • Chapter 5 utilised the methods developed in chapters 3 and 4 to investigate the organisms and genes involved in the degradation of naphthalene vs. methylnaphthalenes in a mixed enrichment culture inoculated with P AB contaminated soil from a former Belfast gas works site. The aim of these experiments was to demonstrate that 2H DNA SIP could be used to gain new information on methylated P AB microbiology and be used specifically for the isolation of biocatalytic genes. • Chapter 1 Presents an introduction to this research • Chapter 2 describes the materials and methods used during the course of this research. • Chapter 6 summarises the work as a whole, suggests future work and discusses the implications of this research in the wider scientific community. • Chapter 7 References |
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