Part:BBa_K1907005

CCP1 promoter for S. cerevisiae

Introduction

CCP1 is a gene for cytochrome-c peroxidase. Cytochrome-c peroxidase is an enzyme that degrades reactive oxygen species in mitochondria. It is also involved in the response to cell’s oxidative stress. As Ccp1p is produced under oxidative stress, its promoter is activated in these conditions. (Saccharomyces genome database ID: S000001774)

This part contains the promoter region (756 bp) of the CCP1 gene. The length of promoter region is chosen to be the same as previously used by He et al. (2005). This promoter length contains all identified Skn7 and Yap1 binding sites and doesn’t overlap with the next gene in the genome. Skn7 and Yap1 are the main transcription factors activated in oxidative stress and are thus responsible for CCP1 promoter activation. The gene sequence for the CCP1 promoter region is obtained from strain S288C of Saccharomyces cerevisiae, from the Saccharomyces Genome Database.

The functionality of this promoter has been tested in association with the Venus YFP reporter in S. cerevisiae strain SS328-leu, and proven to be functional when oxidative stress is induced by hydrogen peroxide. The CCP1 promoter is hardly expressed in a normal state, and although the expression levels don’t rise very high when induced, the difference in expression is very significant.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 290
  Illegal SpeI site found at 364
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 290
  Illegal SpeI site found at 364
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 453
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 290
  Illegal SpeI site found at 364
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 290
  Illegal SpeI site found at 364
  Illegal NgoMIV site found at 613
  Illegal NgoMIV site found at 623
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 158
  

Please find the composite part composed of the CCP1 promoter and Venus YFP, which we have used to study the behavior of the promoter, here: https://parts.igem.org/Part:BBa_K1907008

For more details on the experiments done with this BioBrick, visit http://2016.igem.org/Team:Aalto-Helsinki/Laboratory

References

He, X.J. and Fassler, J.S., 2005. Identification of novel Yap1p and Skn7p binding sites involved in the oxidative stress response of Saccharomyces cerevisiae. Molecular microbiology, 58(5), pp.1454-1467.