Signalling

Part:BBa_I746008

Designed by: Lovelace Soirez   Group: iGEM07_Cambridge   (2007-10-23)

I0500 (Inducible pBad/araC) + I746001 (AIP generator - agrB + agrD with RBSes + Terminator)

This part is the AIP generator (I746001) from the agr system with an arabinose-inducible promoter (I0500) upstream.

pBad is an E. coli promoter that is tightly controlled by:

  • inducer: L-[http://openwetware.org/wiki/Arabinose arabinose].
  • repressor: AraC apparently acts as the repressor

The original (I746000) part was made in order to transfer the S. aureus oligopeptide-based quorum sensing system into a BioBrick-compatible signalling mechanism. It contains the coding regions for group I agrB and agrD (in that order) from S. aureus strain NCTC8325, prefixed by the B0034 RBS and separated by a SaII site.

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.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 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


[edit]
Categories
Parameters
None