Part:BBa_K216013
TNT receptor TNT-R1 + hybrid signal transduction protein Trz
This consists of the coding sequences for TNT receptor TNT-R1 (made by rational redesign of ribose binding protein by the Hellinga et al, Nature 2003) and the hybrid signal transduction protein Trz (consisting of the extracellular part of chemotaxis receptor Trg fused to the intracellular part of sensor kinase EnvZ, which phosphorylates OmpR, controlling promoters such as PompC; made by Hazelbauer et al).
Usage and Biology
This part represents a TNT (2,4,6-trinitrotoluene) detection system. TNT should interact with the modified Ribose Binding Protein TNT-R1, and this should then interact with the extracellular domain of the hybrid Trz protein, initiating kinase activity of the intracellular EnvZ domain, which will phosphorylate OmpR, leading to activation of promoters such as PompC. Thus, a reporter gene attached to PompC (available as BBa_R0082), such as our part BBa_K216011, should be induced in the presence of TNT. Our initial experiments have shown a high basal activity of promoter PompC (BBa_R0082), presumably due to activation by native EnvZ, which responds to osmotic stress (as well as artificial ligands such as procaine). We therefore recommend that an envZ mutant chassis should be used. We have prepared such a chassis, by transduction from the KEIO collection, and will post further information when we have tested it.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 372
Illegal NgoMIV site found at 494
Illegal NgoMIV site found at 624
Illegal AgeI site found at 1224 - 1000COMPATIBLE WITH RFC[1000]
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