BBa_K212001

BBa_K212001 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K212001
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Elias Scheer
Date created: 2009-10-18 11:00:00
Date modified: 2015-05-08 01:11:29

Tar-Envz (Taz)



Types
DnaRegion

Roles
Coding

CDS

Sequences BBa_K212001_sequence (Version 1)

Description

In our system, the Tar-EnvZ (or Taz) chimera protein is used to indicate and signal the presence of aspartate as a chemical ligand. The native form of Tar, found in E. coli, has three domains: a periplasmic ligand binding receptor domain, a transmembrane domain, and an intracellular kinase domain. When aspartate binds to the receptor domain, the kinase domain subsequently transduces a message by phosphorylating CheA and CheY, which orients the bacteria's chemotaxis machinery (i.e. flagellae) towards the aspartate source. In the Taz construct, however, the kinase domain is omitted and the receptor and signal transduction domains of Tar are ligated to the kinase domain of EnvZ, natively an osmolarity sensor kinase-phosphotransferase. Through the kinase activity of EnvZ, this chimeric protein phosphorylates transcription factor OmpR, which subsequently activates transcription of the OmpC gene (or whatever is under the OmpC promoter).

Notes

This is a chimeric two-component transduction system, so we had to be careful to avoid cross-talk by making sure that EnvZ phosphorylates OmpR only.

Source

Tar-EnvZ (Taz) is a chimera protein, manufactured by Inouye, et al. We obtained the gene from his lab and Biobricked it. Taz comprises of the aspartate chemoreceptor region of Tar, the transmembrane region of Tar, and the intracellular kinase region of EnvZ. The genes were fused by digesting both with NdeI and ligating the overlapping ends together. The cut site is between amino acids H256 and M257.

igem#experience
None
 
igem#sampleStatus
It's complicated
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K212001/1