Composite

Part:BBa_K3610031:Design

Designed by: Jonas Sebastian Trottmann   Group: iGEM20_UZurich   (2020-09-27)


BAK1 without native signal peptide / YFP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1758
    Illegal PstI site found at 848
    Illegal PstI site found at 893
    Illegal PstI site found at 1183
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1758
    Illegal PstI site found at 848
    Illegal PstI site found at 893
    Illegal PstI site found at 1183
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1758
    Illegal PstI site found at 848
    Illegal PstI site found at 893
    Illegal PstI site found at 1183
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1758
    Illegal PstI site found at 848
    Illegal PstI site found at 893
    Illegal PstI site found at 1183
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The receptor coding protein of this part has no original signal sequence. The signal peptide was removed (together with the start codon). Therefore, addition of the correct signal peptide sequence is crucial for expression.

We were able to sequence this part, which confirmed that the sequence we used was actually the correct one.

The 15 amino acid linker was chosen because it consists of small amino acids (all Ser and Gly). There is no additional tag for Western Blotting as stains for YFP are widely available.

Source

Composed of the basic parts: BBa_K319005, BBa_K2809002, BBa_K3610002, BBa_K416001, BBa_J176006, BBa_K2753050

References

Chinchilla, Delphine; Zipfel, Cyril; Robatzek, Silke; Kemmerling, Birgit; Nürnberger, Thorsten; Jones, Jonathan D. G. et al. (2007): A flagellin-induced complex of the receptor FLS2 and BAK1 initiates plant defence. In: Nature 448 (7152), S. 497–500. DOI: 10.1038/nature05999.

Macho, Alberto P.; Zipfel, Cyril (2014): Plant PRRs and the Activation of Innate Immune Signaling. In: Molecular Cell 54 (2), S. 263–272. DOI: 10.1016/j.molcel.2014.03.028.

Yan, Liming; Ma, Yuanyuan; Liu, Dan; Wei, Xiaochao; Sun, Yuna; Chen, Xiaoyue et al. (2012): Structural basis for the impact of phosphorylation on the activation of plant receptor-like kinase BAK1. In: Cell Res 22 (8), S. 1304–1308. DOI: 10.1038/cr.2012.74.