A Protocol of Using White/Red Color Assay to Measure Amyloid-induced Oxidative Stress in Saccharomyces cerevisiae

Bharathi, V and Girdhar, A and Patel, Basant Kumar (2017) A Protocol of Using White/Red Color Assay to Measure Amyloid-induced Oxidative Stress in Saccharomyces cerevisiae. Bio-Protocol, 7 (15). ISSN 2331-8325

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The yeast Saccharomyces cerevisiae (S. cerevisiae) harboring ade1 or ade2 mutations manifest red colony color phenotype on rich yeast medium YPD. In these mutants, intermediate metabolites of adenine biosynthesis pathway are accumulated. Accumulated intermediates, in the presence of reduced glutathione, are transported to the vacuoles, whereupon the development of the red color phenotype occurs. Here, we describe a method to score for presence of oxidative stress upon expression of amyloid-like proteins that would convert the red phenotype of ade1/ade2 mutant yeast to white. This assay could be a useful tool for screening for drugs with anti-amyloid aggregation or anti-oxidative stress potency.

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IITH Creators:
IITH CreatorsORCiD
Patel, Basant Kumarhttp://orcid.org/0000-0001-9465-4803
Item Type: Article
Additional Information: This protocol is adapted from our original manuscript published previously as Bharathi et al . (2016). We thank Prof. Susan Liebman, University of Nevada Reno, USA, for yeast strains and plasmids. We thank IIT -Hyderabad, funded by MHRD, Government of India, for consumable support and research infrastructure. Junior Research Fellowship to Vidhya Bhar athi from DBT, Government of India, and Senior Research Fellowship to Amandeep Girdhar from MHRD, Government of India, are also being duly acknowledged.
Uncontrolled Keywords: Amyloid, Oxidative stress, ROS, Yeast, ade1 mutant, TDP-43, DCFDA
Subjects: Others > Biotechnology
Divisions: Department of Biotechnology
Depositing User: Team Library
Date Deposited: 24 Aug 2017 06:36
Last Modified: 10 Nov 2017 06:35
URI: http://raiith.iith.ac.in/id/eprint/3499
Publisher URL: https://doi.org/10.21769/BioProtoc.2440
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