Regulatory

Part:BBa_K3930014

Designed by: Thomas Gaudin   Group: iGEM21_Toulouse_INSA-UPS   (2021-10-08)
Revision as of 08:26, 16 October 2021 by Brice (Talk | contribs)


Doxycycline inducible promoter Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 282
    Illegal XhoI site found at 239
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Introduction

The promoter TetO7 is an doxycycline-inducible promoter. It must be used with the part coding for the activator protein rtTA advanced (BBa_K3930019). The sequence comes from the publication (Garí et al. 1997)

Results

Production of β-carotene (BBa_K3930003)

The part (BBa_K3930002) was linearized and transformed into the S.cerevisiae LycoYeast strain. The production of β-carotene was measured by HPLC. Figure 1 shows the chromatogram of a induced strain with 10 μg.ml-1 of doxycycline.



Figure 1: Production of β-carotene upon doxycycline activation

β-caroteneis produced in vivo by our strain. On the right are presented the mass spectra that correspond between the standard and the observed peak


This promoter Teto7 isn't negatively regulated and is always actived under those lab conditions.


References

  1. Garí E, Piedrafita L, Aldea M, Herrero E. 1997. A set of vectors with a tetracycline-regulatable promoter system for modulated gene expression in Saccharomyces cerevisiae. Yeast. 13(9):837–848. doi:10.1002/(SICI)1097-0061(199707)13:9<837::AID-YEA145>3.0.CO;2-T.


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