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Characterisation of the effects of mutants in the integral dynamics of glutathione systems in regulating oxidative stress in Saccharomyces cerevisiae

Characterisation of the effects of mutants in the integral dynamics of glutathione systems in regulating oxidative stress in Saccharomyces cerevisiae

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An essay for the 2012 Undergraduate Awards Competition by Dean Rowe. Originally submitted for Biological and Biomedical Sciences , with lecturer Gary Jones in the category of Life Sciences
An essay for the 2012 Undergraduate Awards Competition by Dean Rowe. Originally submitted for Biological and Biomedical Sciences , with lecturer Gary Jones in the category of Life Sciences

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Published by: Undergraduate Awards on Aug 30, 2012
Copyright:Attribution Non-commercial

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10/27/2013

 
Title:
Characterisation of the effects of mutants in theintegral dynamics of glutathione systems in regulatingoxidative stress in
 Saccharomyces cerevisiae
1
 
 DECLARATION 
I declare that this thesis is my own work and has not been submitted inany form for another degree or diploma at any university or other institution of tertiary education.Information derived from the published or unpublished work of othershas been acknowledged in the text and a list of references is given.
2
 
Table of ContentsAbstract5Section 1: Introduction71.1: Eukaryotic oxidative stress
71.1.1: Eukaryotic oxidative stress and related mammalian disease71.1.2: Yeast as a model organism 81.1.3: Eukaryotic oxidative stress regulating systems81.1.4: Glutathione discovery91.1.5: Glutathione: Biosynthesis, function and location91.1.6:
GSH1
and
GSH2
role in eukaryotic cells10
1.2: Essential redox regulating systems in
 S. cerevisiae
10 
1.2.1: Glutathione peroxidase systems101.2.2: Glutaredoxin systems 111.2.3: Glutathione transferase systems121.2.4: Omega class glutathione transferases systems131.2.5: Thioredoxin systems 131.2.6: Met4 transcription factor activity14
1.3: Complexities within yeast redox regulatory systems14
1.3.1: Yeast redox regulating protein compensations and multiplicities14
1.4: Aims and Objectives16Section 2: Materials and Methods17 
2.1: Yeast strains used in this work172.2: Reagents182.3: Media and Growth Conditions182.4: Viability determination192.5: Replica plate production192.6: Strain induced sporulation202.7: Random spore analysis202.8: Potential double knockout analysis20
Pilot Experiments:21
2.9: PCR primer design for potential
GPX1
/
Δ
GPX2
double knockout
Δ
21
2.10: Yeast protein extraction 212.11: Bradford protein assay222.12: Superoxide dismutase assay23 3

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