IGEM:Paris Bettencourt 2012/Notebooks/suicide group

From OpenWetWare

Jump to: navigation, search

The aim of this subproject is to design an effective killing mechanism for the bacteria. We are designing this mechanism using Colicin toxin-antitoxin system.

Contents

Colicins

Image:Cliché_2012-06-26_12-16-41.jpg

Image taken from: Cascales E, et al. (2007) Colicin biology. Microbiol Mol Biol Rev71:158–229.

Experiments

We plan to do some experiments to test if our system works and characterize it.

  1. Toxin+antitoxin vs wildtype: We will compete two strains of E coli, one which contains the toxin+antitoxin and is tagged with GFP, and one wild type strain that is tagged with RFP. If our system is working, we expect to the GFP tagged cells to quickly dominate and no RFP tagged cells to remain.
  2. Toxin, antitoxin vs wildtype: We will compete two strains of E coli, one with toxin and antitoxin ion two different plasmids and the wild type. We will compare this result with the result from previous experiment.
  3. Toxin, antitoxin vs antitoxin: We will compete two strains of E coli which are the one with toxin and antitoxin, and the one with only antitoxin. Using fluoscence tag we expect that the strain with only antitoxin will win.
  4. Calibration: We will vary the concentration of inducers of both the toxin and antitoxin promoters to see what is the optimum condition for our system to work.

Parts

Colicins and colicins-related

  • BBa_K131000 Colicin E2 operon, designed by Kevin McLeod, group: iGEM08_Calgary_Wetware (2008-07-22)
  • BBa_K117001 Colicin E7 with immunity, designed by Nguyen Xuan Hung, group: iGEM08_NTU-Singapore (2008-10-07)
  • BBa_K117000 Lysis gene (promotes lysis in colicin-producing bacteria strain), designed by Nguyen Xuan Hung, group: iGEM08_NTU-Singapore (2008-10-07)
  • BBa_K117009 colicin E7 production system induced by lactose, designed by Nguyen Xuan Hung, group: iGEM08_NTU-Singapore (2008-10-08)

Promoters

  • BBa_R0040 TetR repressible promoter, designed by Designed by June Rhee, Connie Tao, Ty Thomson, Louis Waldman, group: Registry (2003-01-31)
  • BBa_R0011 Promoter (lacI regulated, lambda pL hybrid), designed by Neelaksh Varshney, Grace Kenney, Daniel Shen, Samantha Sutton, group: Registry (2003-01-31)

Modeling

References

  • Cascales E, et al. (2007) Colicin biology. Microbiol Mol Biol Rev71:158–229.
Personal tools