Part:BBa_K818400

AmilCP with sboA regulator

This is a coding construct for B. subtilis to produce AmilCP pigment in response to rotten meat volatiles in the environment. AmilCP is a blue pigment designed by iGEM Uppsala 2011 and the regulator, sboA, is designed by iGEM Groningen 2012.

Usage and Biology

Expression in E. coli SboA-AmilCP is strongly expressed in E. coli, on plate and in liquid culture, at normal growth conditions. On plate, the blue-purple colour is less visible compared to the cell pellet in liquid culture.

Expression in B. subtilis

SboA-AmilGFP was shown to be very weakly expressed in Bacillus subtilis on LB plate without rotten meat induction (faint color formation after 4 days). This is probably due to the leakiness of the promoter.

We (iGEM Groningen 2012) tested the expression of SboA-AmilCP in Bacillus subtilis subjected to volatiles from spoiled meat using the same setup as we used to test the sticker design (see our sticker page). After this setup, a clear blue-purple color was visible.

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 207
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]