Part:BBa_I746107

I746101 (AIP sensor infrastructure) + I746105 (Inducible P2 + GFP reporter)

This part is the AIP sensor infrastructure (I746101) from the agr system with an AIP-inducible promoter P2 and GFP reporter (I746105) downstream.

A constitutive/inducible promoter may be added upstream of this part, which will cause production of the components of the signal transduction pathway (AgrC and AgrA); when phosphorylated AgrA is present, the AIP-sensitive promoter (P2) is activated and GFP (E0840) is output.

The original part was made in order to transfer the S. aureus oligopeptide-based quorum sensing system into a BioBrick-compatible signalling mechanism.

In the natural system, the signalling oligopeptide (termed AIP) is made from AgrD by the membrane-located enzyme AgrB. It is then detected by the membrane-located AgrC, which phosphorylates AgrA which then has DNA-binding activity and upregulates transcription of the promoters termed P2 and P3 in the agr locus. There are four known variants of AIP with different molecular structures and cross-inhibitory activity; this BioBrick generates group I AIP.

The agr P2 operon is an autocatalytic sensory transduction system in Staphylococcus aureus. P2 promoter regulates the synthesis of AgrB (a transmembrane protein) and AgrD (precursor of AIP autoinducing peptide). AIP binds to AgrC, a membrane binding receptor and it phosphorylates AgrA. The phosphorylated AgrA increases the activity of agr P2 promoter about 50-fold.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 383
  Illegal BamHI site found at 1267
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2834