Emmatyrnauer Week 12

Journal Outline

Introduction

• Ambient temperature affects gene regulation
• Has led to the evolution of biochemical and physiological properties
• Known cellular activities in response to:
  • heat shock: heat shock proteins expressed more
  • cold shock: decreased permeability of cell membrane, decreased protein synthesis and folding, and decreased enzymatic activity
• Currently there aren't any studies that examine the effects of cold shock below 10 deg C on gene expression in yeast
• Interested in this because preservation of organisms often occurs at less than 10 deg C (i.e. 4 deg C)
• This study exposed yeast to temperature of 4 deg C and examines changes in gene expression using cDNA microarray
  • Cluster analysis performed
    • Results include gene expression between 6 and 48 hours after 4C cold shock differs from culture kept at 35C or 4C. After cold shock, expression of certain genes was induced (e.g. energy preservation genes, phospholipid synthesis genes, and cell rescue genes)
• Now the question is what are the cellular responses of yeast which have already adapted to/able to survive at 4C

Materials and Methods

1. Strain and growth conditions
   • Saccharomyces cerevisia grown in YPD medium at 25C in a shaker.
   • One group was left until growth reached late log phase--control group
   • Other group removed once growth reached mid log phase and transferred to 0C.
     • Upon reaching 4C, yeast was flash frozen and stored at -80C
2. RNA was extracted and poly A mRNA was purified
3. Probe prepared and labeled
4. Microarray hybridized and scanned
5. Hierarchal cluster analysis performed with GeneSpring

References

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