Part:BBa_K1486048

CpxR reporter

Calgary's BBa_K339007 [1] repaired: the part didn't have the sequence they said they had, it was missing most of the promoter.

Biology of the Part

E coli have several native sigma factors in the genome that are upregulated during envelope stress. There is also a new regulon that is currently studied widely, called the Cpx regulon, a well characterized periplasmic regulon found in E. coli. The Cpx pathway is involved in maintaining cellular adhesion and keeping periplasmic environment in check. It regulates proteins which are responsible for cellular adhesion and pili formation. The Cpx pathway is activated by factors which cause protein aggregation in the periplasmic space of gram negative bacteria. Extracellular stresses such as exposure to ethanol, change in pH and temperature change induce the activation of the Cpx heat shock regulon. In our BioBrick this should increase the transcription of the RFP which it regulates.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 636
  Illegal AgeI site found at 748
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage

Excitation: 555 nm Emission: 632 nm

Characterisation

To check if the cells were responding to different kinds of stress we put them in different environments: 40 mM KCl, 80 mM KCl, 5 % ethanol and 10 % ethanol . As a negative control we added PBS to the same cells. As another negative control cells that did not have the RFP coding sequence where put in the same environments.

We see that there is no sensitivity to the stress factors in this conditions.