Effects of drugs on yeast signaling pathways

Many drugs are used by people without a full knowledge of drug-induced changes in cell function. In this lab, students will screen two drugs for their effect on various metabolic and signaling pathways. The budding yeast Saccharomyces cerevisiae will be used as a test system because of the wealth of knowledge about this eukaryote. Effects of drugs on yeast signaling pathways will be deduced using from the pattern of whole genome mRNA levels, measured by hybridization to DNA microarrays. Two drugs will be tested: one (an azole drug) with a previously characterized genomic response and a second (??) with an unknown response. Possible candidates for the latter drug include crude chemicals from health food stores:

  1. Grow yeast overnight in YEPD (a rich growth medium)
  2. Dilute ~1 ml O/N culture into 60 mls YEPD (A600=0.1)
  3. Grow cells at 30°C in 250 ml flask, with shaking
  4. After 4 hours, make sure the cells are in log phase (A600 = 0.2)
  5. treat cells for 2 hours as follows:
  6. Isolate total RNA

Copyright, Acknowledgements, and Intended Use
Created by B. Beason (bbeason@rice.edu), Rice University, 14 March 2005
Updated 21 July 2006