Part:BBa_K3962344

arabinose-mediated inducible expression of toxin MazF

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

MazF has been used as a part of a kill switch (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 not inhibited following the induction of MazF. The results show that MazF is not necessarily enough to kill all cells immediately when grown in liquid LB medium. Using minimal medium is likely to produce a different result for this experiment that is more representative of a natural environment. This is because glucose heavily represses the transcription of pBAD. Another factor might be the absence of extracellular death factor (https://doi.org/10.1128/MBIO.00314-13 ).

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