"
Team:McGill/Experimental
From 2009.igem.org
Experimental Notebook
Experiments
Basics
Two separate cells that secrete transcription factors such that they create a feedback system between each other should interact according to the model detailed in the theory section. That is, oscillatory patterns should become apparent. In order to verify the occurrence of oscillations experimentally, we designed two gene constructs that utilize the Rhl quorum sensing system from P. aeruginosa and the Lux system from V. fischeri.
Construct 1
- Secretes 3OC6-HSL in response to C4-HSL
- RhlR constitutively expressed.
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K290003 K290003]
Construct 2:
- Secretes C4-HSL in response to COC6-HSL
- LuxR constitutively expressed.
[http://partsregistry.org/Part:BBa_K290002 K290002]
Plasmid: The constructs as synthesized by GenScript were delivered in a copy-number inducible plasmid to minimize mutations. There seems to be instability due to repeats in the STOP sequences. The plasmid we use is pCC1.
- LINK REGISTRY*
Cells: We use EPI300 cells that were supplied by GenScript.
