Part:BBa_K3610031:Design
BAK1 without native signal peptide / YFP
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000COMPATIBLE 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.