Generator

Part:BBa_K3962343

Designed by: Siheng Li   Group: iGEM21_Leiden   (2021-10-11)


arabinose-mediated inducible expression of toxin RelE

This construct is composed of L-arabinose mediated inducible promoter pBAD and toxin RelE. RelE/RelB is a type II toxin-antitoxin system originating from E. coli. In our project, we used this construct to test the toxicity of toxin RelE, which provided us with more insights in regulating the balance of the toxin-antitoxin system for preventing horizontal gene transfer.


2021 Team Leiden

https://2021.igem.org/Team:Leiden

RelE has been used as a part of a kill switch. The lethality of this kill switch has been tested in a plate reader assay (Fig. 1)

T-Leiden-parts-Figuer5.png

Figure 1. Plate reader assay of E. coli TOP 10 cellular growth over time, with a gradient of arabinose concentrations.

Cell growth is not inhibited following the induction of RelE. The results show that RelE is not enough to kill cells immediately when grown in liquid LB medium.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


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