Part:BBa_K1413023

Promoter-RBS-bphr2 mutated-Terminator

bphR2 gene, from Pseudomonas pseudoalcaligens KF707, allows the transcription of bphR1 which activates the genes of degradation of biphenyls. In our project, genes of degradation was replaced by RFP gene to detect the compound. This part is composed by constitutive promoter (BBa_J23114), RBS (BBa_B0034), bphr2 (BBa_K1413021) and terminator (BBa_B0015) (Figure 1).

Figure 1: Scheme of BBa_K1413023 construction.

Mechanism explanation: In absence of PCBs, bphR2 (BBa_K1413021) is bound to bphR1 promoter which activate the transcription of RFP but in very low expression. In presence of PCBs, when compound diffuses into the media, it binds to bphr2 protein which undergoes a conformational change that permits to activate more stronger bphR1 promoter and increase the transcription of RFP (Figure 2).

Figure 2: Mechanism of PCB biosensor with the system bphR2/bphR1 gene

Usage and Biology

Combinated with the part BBa_K1413024, it could be used like PCB biosensor.

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 568
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 352