Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Daniel Cort??s
Date created: 2016-10-12 11:00:00
Date modified: 2016-10-13 02:23:39
pathogen associated molecular pattern sensor
| Types | DnaRegion |
| Roles | engineered_region Signalling |
| Sequences | BBa_K1909001_sequence (Version 1) |
Description
We created a fusion protein, joining the EnvZ two-component system signalling domain to the sensing leucine-rich repeat domain of the pattern recognition receptor NOD1. The EnvZ kinase domain autophosphorylates after target binding and activates OmpR, a prokaryotic selective transcription factor. Therefore it is able to sense pathogen associated molecular patterns (e.g. LPS, mDAP) and subsequently activate reporter expression.Notes
To prevent premature ribosome drop-off, the pair of CTA codons (L77 and L79 of the LRR domain) predicted to strongly stall the translation was substituted by TTA codons. An XbaI restriction site was removed by changing L239 from CTA to TTA. To make up for the lost translational pause, L242 was changed from CTG to CTA (Fig 1C). We also added an eight aminoacid FLAG-tag at residue 16, allowing for expression confirmation by targeted purification.Source
EnvZ - http://www.uniprot.org/uniprot/P0AEJ4NOD1 - http://www.uniprot.org/uniprot/Q9Y239
| Sequence Annotation | Location | Component / Role(s) |
| FLAG NOD-LRR domain EnvZ Kinase domain | 46,69 169,1135 1136,2011 | feature/tag tag non_covalent_binding_site feature/binding feature/misc sequence_feature |