Part:BBa_K3962341
arabinose-mediated inducible expression of toxin Hok
This construct is composed of L-arabinose-mediate inducible promoter pBAD and toxin Hok. Hok/sok is a type I toxin/antitoxin system originating from E. coli. In our project, we used this construct to test the toxicity of Hok, which provided us with more insights to regulate the toxin-antitoxin system for preventing horizontal gene transfer.
2021 Team Leiden
https://2021.igem.org/Team:Leiden
In our project, we used this construct to test the toxicity of the Hok toxin, which provided us with more insights in regulating the balance of the toxin-antitoxin system for preventing horizontal gene transfer. The toxicity of this kill switch has been assessed via a plate reader assay (Fig 1.).
Figure 1. Plate reader assay of E. coli TOP 10 cellular growth over time with a gradient of arabinose concentrations.
Cell growth is strongly inhibited following the induction of CcdB. The results show that CcdB is not necessarily enough to kill all cells immediately when grown in liquid LB medium. Only after the exponential growth phase, the cells start to die. High concentrations of arabinose are needed for this killing effect, but that might change in different medium conditions.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
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