Team:KUAS Korea/Experiments
Experiments |
1. Designing and constructing the cooperator and cheater
- We have looked the previous iGEM projects and articles on the game theory. We decided to express GFP for cheater and beta-glucosidase for cooperator. We received the designed constitutive expression vector applicable to ligation independent cloning using the plasmid containing BBa_J23106 from Ph.D. Ko.
2.Transformation into E.coli BW25113
- As the E.coli BW25113 is a host strain for the surface display and expression, we transformed the designed plasmid vector into it. We cultured the bacteria on M9 minimal media; one with glucose and another with cellobiose respectively. We checked the activity of cheater on M9 minimal medium with glucose and cooperator on M9 minimal medium with cellobiose.
3.Co-culture of cooperator and cheater with different ratio
- Both cooperator and cheater were incubated in M9 minimal medium with 0.48% glucose at 37’C until it reached 1.0 in OD 600.
- Batch co-culture was conducted in 5ml of M9 minimal medium with 0.48% cellobiose solution; in ratio of cheater and cooperator 1:1, 1:2, 1:4, 2:1, 4:1.
- After 2, 4 and 6 days, each co-culture medium with different ratio was extracted and streaked on LB agar plate with ampicillin.
- After overnight growth, total cell was counted and the plate was checked under fluorescence microscopy to count cheater; GFP expressing bacteria.
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