IGEM:Paris Bettencourt 2012/Notebooks/RE group/Promoter Candidates

Our main objective is to find inducible promoters that are not leaky.

Pbad

• Induced by: Glucose
• Inhibited by: Arabinose

Overview of PBAD Promoter Family: [1]

• Pbad Part:BBa_I13453
• Pbad+AraC Part:BBa_I0500
• Characterizationby the Cambridge 2009 team of different Pbad/araC promoters
• Plasmid: Part pSB3K3

References

1. «In Vivo Induction Kinetics of the Arabinose Promoters in Escherichia coli» Casonya M. Johnson And Robert F. Schleif* Read It

Plac

• Repressed: LacI
• Induced: Lactose, IPTG
• LacIq Part:BBa_K091111
• LacI Part: BBa_C0012
• Lac Operon Part:BBa_R0010
• Lac Operon Part(mutant of the BBa_R0010 part. Lower leakage, but also lower expression) BBa_K611024

References

1. «Tightly regulated vectors for the cloning and expression of toxic genes» Larry C. Anthony*, Hideki Suzuki, Marcin Filutowicz Read It

Ptet

• Activity:
  • Induced by: aTc
  • Repressed by: tetR

• Sequence:
  1. Ptet Part:BBa_R0040

     Sequence for pTet inverting regulator. Promoter is constitutively ON and repressed by TetR. TetR repression is inhibited by the addition of tetracycline or its analog, aTc.

  2. TetR Part:BBa_C0040

     Coding region for the TetR protein without the Ribosome Binding Site. Modified with an LVA tail for rapid degradation of the protein and faster fall time for the emission. TetR binds to the pTet regulator (Part:BBa_R0040). aTc (anhydrotetracycline) binds to TetR and inhibits its operation.

• Caracterization:
  • EPF-Lausanne 2011 team

References

1. «Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements» Rolf Lutz and Hermann Bujard* Read It

pRHA (P Rhamnose)

• Activity:
  • Induced by: L-Rhamnose
  • Inhibited by: D-Glucose
• Sequence:
  1. Prha: Rhamnose->PoPS: BBa_K564001

     This part contains the rhamnose promoter and its two coregulators, rhaS and rhaR. This promoter is activated in the presence of L(+)-Rhamnose and inhibited by glucose due to catabolite repression. It can be used in a very analogous way to the arabinose promoter (i0500).

References

1. «Toxic protein expression in Escherichia coli using a rhamnose-based tightly regulated and tunable promoter system» Matthew J. Giacalone1, Angela M. Gentile2, Brian T. Lovitt2, Neil L. Berkley2, Carl W. Gunderson1, and Mark W. Surber2 Read It
2. «DNA-Dependent Renaturation of an insoluble DNA binding Protein. Identification of the RhaS Binding Site at rhaBAD» Susan M.Egan and Robert F. Schleif Read It
3. «Optimization of an E. coli L-rhamnose-inducible expression vector: test of various genetic module combinations» Angelika Wegerer, Tianqi Sun and Josef Altenbuchner Read It.

   Note: Only need to express the PRhaBad promoter. The other genes are on the E.coli chromosome

Interesting links

• Promotr measurment protocol from Imperial 2009 Read It
• Rhamnose-inducible promoter that drives expression of I-SceI endonuclease [2]