Difference between revisions of "Team:Hong Kong HKUST/pTet"

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<p style="color: black; font-size: 1.2em;"> The <i>tetp</i> module consists of a Tet repressible promoter upstream of the CDSs of phlf and lacI, which are then followed by the reporter gene, mtagbfp. When the ptet module is active, the two repressor proteins, PhlF and LacI will be expressed along with the reporter gene, mTagBFP. This results in the repression of the two other modules in our system, <i>pphlf</i> and <i>plac</i>. </p>
 
 
 
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Revision as of 04:36, 12 October 2016

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tetp Module



The tetp module consists of a Tet repressible promoter upstream of the CDSs of phlf and lacI, which are then followed by the reporter gene, mtagbfp. When the ptet module is active, the two repressor proteins, PhlF and LacI will be expressed along with the reporter gene, mTagBFP. This results in the repression of the two other modules in our system, pphlf and plac.

Promoter, Repressor and Inducer Specification


tetp promoter
  • Promoter name: tetp (Tet repressible promoter)
  • Length: 54 base pairs
  • Strength with:
    • Weak RBS (BBa_B0032): 5.6 - 517.5 (+17) RPU
    • Medium RBS (BBa_B0030): 25.5 - 2323.1 (+259.5) RPU
    • Strong RBS (BBa_B0034): 39.6 - 4041.5 (+247.8) RPU

The tetp module consists of a Tet repressible promoter upstream of the CDSs of phlf and lacI, which are then followed by the reporter gene, mtagbfp. When the ptet module is active, the two repressor proteins, PhlF and LacI will be expressed along with the reporter gene, mTagBFP. This results in the repression of the two other modules in our system, pphlf and plac.

  • Repressor name: TetR (Tet repressor)
  • Part Length: 685 base pairs
  • Structure: Homodimer of alpha-helices
  • Binding affinity: To tetp, very light, 0-1 µl/ml has been shown to cause a 5 order of magnitude change in fluorescent protein production
  • Inducer name: ATc (Anhydrotetracycline)
  • Binding Affinity: (to TetR) Ka ~ 109 M-1
  • Effective range: 250 - 25000 ng/ml

Mechanism of tetp - TetR - ATc Interaction