Team:Warsaw/Stage2/Design

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Design

Design and measurement of kill-switch safety system

The safety system we designed and measured consists of the parts specified: T7 promoter + B0032 RBS + MinC on pSB1A2 plasmid, studied in E.coli BL21 RIL strain.

pT7
I719005

B0032

K299806

We measured our construct in vivo, in the expression system, using strong T7 promoter, IPTG induced (see results section). The functionality of our safety system describes optical density of E.coli cells (OD) and the number of colony forming units per mililiter (cfu/ml) in IPTG-induced innoculates BL21 carrying our construct.

Additional experiment to kill-switch safety system

We also performed experiment with B0034 RBS as an alternative to B0032. We designed biobrick presented below: T7 promoter + B0034 RBS + MinC on pSB plasmid, studied in E.coli BL21 strain.

pT7
I719005

B0034

K299806
As we know from RBS measurement data, B0034 is stronger than B0032. As a result, we didn’t obtain any visible bacterial colonies on agar plates (see results). Our conslucion to this experiment is that additional regulation of MinC gene expression is very important to keep the kill-switch system working.