Coding

Part:BBa_K212001:Design

Designed by: Elias Scheer   Group: iGEM09_Brown   (2009-10-19)
Revision as of 04:25, 20 October 2009 by Escheer (Talk | contribs) (→‎References)

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Tar-Envz (Taz)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 420
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 420
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 420
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 111


Design 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.

References

1.

2. Zhu, Yan, and Masayori Inouye. "Analysis of the Role of the EnvZ Linker Region in Signal Transduction Using a Chimeric Tar/EnvZ Receptor Protein, Tez1." The Journal of Biological Chemistry 278.25 (2003): 22812-2819. Print.