Part:BBa_K1413024

bphR1-RBS-RFP-Terminator

This part is composed by bphR1 promoter (BBa_K1155001), from Pseudomonas pseudoalcaligenes KF707, which allows the transcription of the genes of biphenyl degradation, the RBS (BBa_B0034), RFP gene (BBa_E1010) and terminator (BBa_B0015) (Figure 1). In our project, genes of degradation was replaced by RFP gene to detect the compound.

Figure 1: Schema of BBa_K1413024 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_K1413023, it could be used like PCB biosensor.

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 156
  Illegal BamHI site found at 239
  Illegal XhoI site found at 46
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 883
  Illegal AgeI site found at 995
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 139