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Title: Construction and application of a luminescent eukaryotic biosensor
Author: Hollis, Roger Paul
Awarding Body: University of Aberdeen
Current Institution: University of Aberdeen
Date of Award: 1999
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Accurately measuring in vivo toxicity of chemicals is a demanding task, as it is influenced by elaborate multifactorial systems within and around cells. Existing methods for assaying in vivo toxicity to eukaryotic cells exploit whole animal models or tissue culture, which are time consuming, expensive and raise ethical issues. The aim of this investigation was to create an alternative approach to current cytotoxicity analyses by applying eukaryotic microbial biosensors. The yeast Saccharomyces cerevisiae was genetically modified to express firefly luciferase to create a bioluminescent yeast strain. Bioluminescence responds quantitatively to the presence of any toxic chemical that interferes with the cells' metabolism. S. cerevisiae has been researched extensively to investigate the many homologous cellular systems shared with higher eukaryotes, such as mammalian cells. This homology allows the indicative analysis of compounds for their toxic effects to eukaryotes. In this study it was demonstrated that the luminescent yeast strain senses chemicals known to be toxic to eukaryotes in samples assessed clean by similar prokaryotic biosensors. As the cell wall and adaptive mechanisms of S. cerevisiae cells enhance stability, protect from extremes of pH, solvent exposure and osmotic shock, these inherent properties were exploited to generate a biosensor that can detect a wide range of both organic and inorganic toxins in extreme conditions. Therefore, the novel biosensor developed during this provides a rapid and robust strategy for indicating the toxicity of substances to eukaryotic cells. Such a system could be applied to the routine screening of pharmaceuticals and potential carcinogens, reducing the need for animal testing.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available