Part:BBa_K2120005

araC+PBAD+B0031+hokD

Arac-pBAD is an arabinose induced promoter, hokD is a toxin gene. This part is functioned as a killer.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 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
  

Functional Parameters

This is a part of 2016 BIT-China iGEM Team to verify the function and character of the mazF toxin gene. And it also the verification circuit of the killer device in the project. We got the circuit by using one-step mutation from araC+PBAD+B0032+hokD BBa_K2120003. And we constructed the plasmid containing araC+PBAD+B0031+mazF circuits, and transformed the plasmid into top10. After transformation, we measured the OD600 to draw the growth curve and observed the function of toxin protein. Besides the bacteria contained the toxin gene, the empty pSB1C3 vector was the control. Add arabinose or not, there was another comparison. We add 10% arabinose 50 uL into 50 mL LB culture medium when the OD600 is 0.6 (the log phase). We measured the OD600 every hour until the bacteria reached the stationary phase.

Fig. Compared with control groups, the difference value of OD600 showed the lethal efficiency of the circuit

According to the results, only the bacteria containing the plasmid with toxin gene and induced by arabinose can be inhibited and killed. For more results, please see: BBa_K2120002(araC+PBAD+B0032+mazF), BBa_K2120003(araC+PBAD+B0032+hokD), BBa_K2120004(araC+PBAD+B0031+mazF), BBa_K2120006(araC+PBAD+B0034+mazF).