IGEM:Paris Bettencourt 2012/Notebooks/suicide group: Difference between revisions
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The aim of this subproject is to design an effective killing mechanism for the bacteria. We | 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. | ||
= Colicins = | = Colicins = | ||
| Line 6: | Line 6: | ||
Image taken from: Cascales E, et al. (2007) Colicin biology. Microbiol Mol Biol Rev71:158–229. | Image taken from: Cascales E, et al. (2007) Colicin biology. Microbiol Mol Biol Rev71:158–229. | ||
= Experiments = | |||
We plan to do some experiment to test if our system works and characterize it. | |||
# '''Toxin+antitoxin vs wildtype''': We will compete two strains of E coli which are the one with toxin+antitoxin in the same plasmid and the wild type. Using fluoscence tag we expect that the strain with toxin+antitoxin will win. | |||
# '''Toxin, antitoxin vs wildtype''': We will compete two strains of E coli which are the one with toxin and antitoxin in the different plasmid and the wild type. We will compare this result with the result from previous experiment. | |||
# '''Toxin, antitoxin vs wildtype''': 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. | |||
# '''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 = | = Parts = | ||
==Colicins and | ==Colicins and colicins-related== | ||
*[http://partsregistry.org/Part:BBa_K131000 BBa_K131000] Colicin E2 operon, designed by Kevin McLeod, group: iGEM08_Calgary_Wetware (2008-07-22) | *[http://partsregistry.org/Part:BBa_K131000 BBa_K131000] Colicin E2 operon, designed by Kevin McLeod, group: iGEM08_Calgary_Wetware (2008-07-22) | ||
Revision as of 03:39, 3 July 2012
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.
Colicins
Image taken from: Cascales E, et al. (2007) Colicin biology. Microbiol Mol Biol Rev71:158–229.
Experiments
We plan to do some experiment to test if our system works and characterize it.
- Toxin+antitoxin vs wildtype: We will compete two strains of E coli which are the one with toxin+antitoxin in the same plasmid and the wild type. Using fluoscence tag we expect that the strain with toxin+antitoxin will win.
- Toxin, antitoxin vs wildtype: We will compete two strains of E coli which are the one with toxin and antitoxin in the different plasmid and the wild type. We will compare this result with the result from previous experiment.
- Toxin, antitoxin vs wildtype: 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.
- 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
- 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)
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)
References
- Cascales E, et al. (2007) Colicin biology. Microbiol Mol Biol Rev71:158–229.
