Part:BBa_K216004

Trz hybrid signal transduction protein

Trz hybrid signal transduction protein: this was created by fusing the extracellular receptor domain of the chemoreceptor Trg with the intracellular signal transduction domain of 2-component sensor protein EnvZ, which normally phosphorylates response regulator OmpR (see Baumgartner, J.W., Kim, C., Brissette, R.E., Inouye, M., Park, C., and Hazelbauer, G.L. 1994. Transmembrane signalling by a hybrid protein: communication from the domain of chemoreceptor Trg that recognizes sugar-binding proteins to the kinase/phosphatase domain of the osmosensor EnvZ. J. Bacteriol. 176, 1157-1163). This part can activate genes attached to an OmpR-responsive promoter such as PompC. To avoid inappropriate activation via EnvZ, it may be necessary to use a chassis in which EnvZ is not present.

Usage and Biology

Trz is a fusion protein with the extracellular region of the chemoreceptor Trg fused to the intracellular kinase domain of two-component sensor EnvZ, an osmosensor which phosphorylates the cognate response regulator OmpR. Phosphorylated OmpR then activates promoters such as the ompC promoter (available as BBa_R0082). Trg normally detects chemoattractants such as ribose, bound to the periplasmic ribose-binding protein. The usefulness of Trz arises in that ribose binding protein can be computationally redesigned to bind non-natural ligands, and this binding can then be detected via actiavtion of a reporter gene fused to the ompC promoter (Looger et al, 2003). Thus this is potentially a generalisable transduction system for detection of extracellular ligands.

Trz was previously deposited as part BBa_J58104 by Valencia iGEM 2006; however, Registry sequence analysis of this part indicates that the sequence is not correct. It appears in fact to be vector DNA or E. coli genomic DNA of the lacI region. This part is therefore a replacement for BBa_J58104. We have confirmed the sequence of our part but at the time of writing have not yet confirmed its activity.

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 365
  
• 1000
  COMPATIBLE WITH RFC[1000]