Part:BBa_K3610004

BAK1 ectodomain from Arabidopsis thaliana - codon optimized for C. reinhardtii

This part a protein coding sequence compromising the ectodomain of the brassinosteroid insensitive 1-associated receptor kinase 1. This part is a codon optimized version of BBa_K3610003 and therefore lacks the intracellular kinase domain and its native signal peptide. It has been codon optimized for the expression in C. reinhardtii.

Usage and Biology

The BRI1-associated receptor kinase (BAK1) is a leucin-rich repeat receptor kinase (LRR-RK) which interacts with multiple other LRR-RKs with different functions in hormone signalling and defense response. BAK1 localizes at the plasma membrane and the endosome. The BAK1 protein forms a structure with an extracellular domain with leucin-rich repeats, a single pass transmembrane domain and an intracellular domain with a kinase function.

Among others, BAK1 interacts with the LRR-RKs EF-Tu receptor (EFR), Flagellin sensing 2 (FLS2) and cold-shock protein receptor (CORE), all of which are pathogen recognition receptors (PRR) in brassicaceae plants. Upon binding of a microbe-associated molecular pattern at the LRR domain of the PRR, BAK1 forms a heterodimer with the PRR which triggers a phosphorylation cascade, leading to upregulation of defense mechanisms.

For visualizing the interaction of BAK1 with other plant receptors the cytoplasmic domain can be replaced with a split fluorescent protein or another protein that generates a visual output. In our iGEM project, we fused this sequence to a split-mCherry protein and a split-luciferase and then coexpressed the part with another plant PRR carrying the counterpart to the split protein instead of the intracellular kinase domain. As we expressed the BAK1 ectodomain also in C. reinhardtii, we created this sequence. Localization at the cell surface in C. reinhardtii is ensured by a signal peptide native to the host organism.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 778
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 708
  
• 1000
  COMPATIBLE WITH RFC[1000]