Difference between revisions of "Part:BBa K1907003"
 
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Please find the composite part composed of the CTT1 promoter and Venus YFP, which we have used to study the behavior of the promoter, here:
 
Please find the composite part composed of the CTT1 promoter and Venus YFP, which we have used to study the behavior of the promoter, here:
 
https://parts.igem.org/Part:BBa_K1907009
 
https://parts.igem.org/Part:BBa_K1907009
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For more details on the experiments done with this BioBrick, visit http://2016.igem.org/Team:Aalto-Helsinki/Laboratory
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Latest revision as of 18:03, 20 October 2016


CTT1 promoter for S. cerevisiae


Introduction

CTT1 is a gene for cytosolic catalase T. This protein has an important role in protecting the cell from oxidative damage caused by hydrogen peroxide. As Ctt1p is produced under oxidative stress, its promoter is activated in these conditions. (Saccharomyces genome database ID: S000003320)

This part contains a promoter region (924 bp) of CTT1 gene. The length of promoter region is chosen so that contains identified Skn7 and Yap1 binding sites and doesn’t overlap with the next gene in the genome. (He et al., 2005) Skn7 and Yap1 are the main transcription factors activated in oxidative stress and are thus responsible for CTT1 promoter activation. In the CTT1 promoter region, these binding sites are rather close to start codon (-147 bp), and as there are no genes close to it, a longer promoter was decided to be used just in case. The gene sequence for the CTT1 promoter region is obtained from strain S288C of Saccharomyces cerevisiae from the Saccharomyces Genome Database.

The functionality of this promoter was tested in association with the Venus YFP reporter in S. cerevisiae strain SS328-leu, with oxidative stress being induced by hydrogen peroxide. Hydrogen peroxide didn’t seem to activate the promoter, but reaching higher cell densities in cell growth did. This promoter could thus be e.g. used in monitoring the growth phase of yeast cells.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 402
    Illegal PstI site found at 695
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 402
    Illegal NheI site found at 701
    Illegal PstI site found at 695
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 402
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 402
    Illegal PstI site found at 695
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 402
    Illegal PstI site found at 695
    Illegal AgeI site found at 555
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 508

Please find the composite part composed of the CTT1 promoter and Venus YFP, which we have used to study the behavior of the promoter, here: https://parts.igem.org/Part:BBa_K1907009


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.