Coding

Part:BBa_K3610008

Designed by: Jonas Sebastian Trottmann   Group: iGEM20_UZurich   (2020-09-08)
Revision as of 19:00, 3 October 2020 by Jtrott (Talk | contribs)

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EFR ectodomain - codon optimized for C. reinhardtii

This part is a codon optimized version of Part:BBa_K3610007. It is optimized for the expression in C. reinhardtii.

Usage and Biology

Elongation factor-thermo unstable receptor (EFR) from A. thaliana is a plant pattern-recognition receptor (PRR). It is a cell surface receptor and part of the plants firts defence mechanism against potential pathogens. The EFR receptor is also a leucin-rich-repeats (LRR) receptor-like serine/threonine-protein kinase. The protein consists of an extracellular domain with leucin-rich repeats, a ligand binding domain found in many receptors, a single-pass transmembrane domain and finally an intracellular kinase domain. The ligand binding domain from EFR has high specificity to a bacterial pathogen-associated moleculat pattern (PAMP), namely the epitope elf18 of the abundant protein Elongation Factor Tu (EF-Tu), which is catalyzes the binding of aminoacyl-tRNA (aa-tRNA) to the ribosome in most prokaryotes and therefore is evolutionarily highly conserved. This makes the EFR a receptor that can be activated by the presence of a huge variety of bacteria. Upon binding of the ligand to the extracellular domain, the receptor dimerizes with its coreceptor BRI1-associated receptor kinase (BAK1). This interaction triggers the activation of the intracellular kinase domain of EFR and BAK1, initiating a signal cascade leading to an upregulation of immune response mechanisms.

This was designed for protein-fragment complementation experiments (PCA). It can be fused to a split protein and coexpressed with BAK1, fused to the counterpart of the split protein, to visualize the interaction between the two receptors. Examples of such a complementation system are split-mCherry and the Split-NanoLuc system. The part has been codon optimized for expression in C. reinhardtii.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 82
    Illegal NheI site found at 1012
    Illegal NheI site found at 1930
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1598
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None