BBa_K777004

BBa_K777004 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K777004
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Bianca Genenncher
Date created: 2012-09-20 11:00:00
Date modified: 2015-05-08 01:13:16

Tar receptor under the control of constitutive promoter J23106



Types
DnaRegion

Roles
Composite

engineered_region

Sequences BBa_K777004_sequence (Version 1)

Description

[[Image:Table_K777001-K777008_new.png|thumb|240px|right|Fig. 1: Overview of BioBricks containing the Tar gene downstream of Anderson promoters. (Activity according to Berkeley 2006)]]
The transmembrane chemoreceptor Tar of E. coli mediates chemotaxis towards aspartate and exists as a functional homodimer.

Here we used a set of 8 constitutive Anderson promoters from the 2006 Berkeley group to test how different levels of constitutive Tar expression affect the chemotaxis of E. coli.

















Notes

  • Genomic sequence was amplified using the following primers.
    • Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
      • Fwd: Tar for + prefix: 5?? actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3??
      • Rev: Tar rev + suffix: 5?? tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3??



  • One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR.
    • Primers QuikChange (QC): Primers were provided by SIGMA.
      • Fwd: TarQC for: 5?? ACTGATTGATTAcCTAGATTATGGCAATACTGGAG 3??
      • Rev: TarQC rev: 5?? TGCCATAATCTAGgTAATCAATCAGTTCAGTTAAC 3??
        (the uncapitalized letter induces the mutation for removal of the XbaI site)


Source

*The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B, complete genome (CP000948.1).
*J23106 information was taken from the partsregistry and physical DNA from the 2012 distribution kit.

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