Part:BBa_K3736001

constitutive promoter+ FourU + luxR + Terminator*2

constitutive promoter+ FourU + luxR + Terminator*2

Quorum sensing sequence in DenTeeth

   The RBS of BBa_K3736001 (Quorum sensing sequence) needs to exceed 37 degrees to enable its mRNA to begin translation and initiate quorum sensing regulation. In this way, we can ensure that our engineered bacteria can operate in a specific temperature range. When it develops into a commodity in the future, it can also reduce the production of antimicrobial peptides before entering the body of a warm-blooded animal, making it stable and easy to store.

   When the LuxR protein produced by BBa_K3736001 (Quorum sensing sequence) binds to the AHL produced by all bacteria, it can prompt the Plux promoter of BBa_K3736002 (Inhibition sequence) to start transcription, so that BBa_K3736002 (Inhibition sequence) can be expressed when there are more bacteria. When the tetR protein in BBa_K3736002 (Inhibition sequence) is expressed, the Ptet promoter of BBa_K3736003 (Restoration sequence) will be inhibited to achieve the antagonistic effect of BBa_K3736002 (Inhibition sequence) and BBa_K3736003 (Restoration sequence).

Figure 1. Quorum sensing sequence

Gene Construct of DenTeeth

  We incorporate the whole part into E. coli BL21(DE3). We did colony PCR and digest to check its genotype.

Figure 2. Quorum sensing sequence + Restoration sequence + Sterilization sequence. M :1kb DNA ladder, 1 : Constitutive promoter + RBS + LuxR + Term. +Term. + Ptet + RBS + BMP2 + RBS + STATH + RBS + GFP + Term. + Term. + Plux + RBS + LL-37 + RBS + mRFP + RBS + tetR + Term. + Term. (4685 b.p.)</i>

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 890
  Illegal NgoMIV site found at 1011
  
• 1000
  COMPATIBLE WITH RFC[1000]