• Introduction
  • Home
  • The Team
  • Photo Gallery
• Our Project
  • Results Summary
  • Selection System
  • In-Vitro Characterization
  • In-Vivo Characterization
  • Microfluidics
  • Data
  • Attributions
• Tools
  • Gibson Assembly
  • MITOMI
• Notebook
  • Protocols
  • May
  • June
  • July
  • August
  • September
  • October
• Considerations
  • Human Practices
  • Safety
• Acknowledgements
  • Sponsors
  • Partners

Reporter systems

We built two reporter systems in order to characterize and select our transcription factors in vivo. The first reporter gene is the lysis cassette, allowing to recover DNA from the interesting mutants. The second gene is RFP, for characterizing more precisely the mutant TetR-Ptet interaction.

As the lysis device has been characterized in the T7 promoter variants section, we focused here on RFP induction. The following is referring to RFP as the only reporter gene, but we could easily use our systems for lysis induction by exchanging RFP for hte lysis cassette.

TetR-RFP system

The first system is the simplest one; it is composed of TetR driven by a constitutive promoter and of RFP under Ptet control. If TetR binds to Ptet, then RFP is repressed. This readout system is convenient for fluorescence detection experiments, but it would not be suited for using the lysis cassette as the reporter gene is being repressed upon TetR-Ptet interaction. With the lysis device, the interesting cells (where TetR binds to Ptet) would survive and we would recover only the useless TetR mutants.