Part:BBa_K3610050

CORE ectodomain / mCherry C-terminal for C. reinhardtii

This part includes the ectodomain of the plant pattern recognition receptor CORE fused to the N-terminal domain of the split-mCherry protein. The sequence for the receptor and the mCherry protein have been codon optimized for expression in C. reinhardtii. To ensure localization at the membrane, this part further contains the sequence for the signal peptide SP7 from C. reinhardtii and the self-cleaving protein from the foot and mouth virus.

Usage and Biology

CORE

The cold shock protein receptor (CORE) is a plant pattern recognition receptor (PRR) and as such activates host innate immunity through detection of pathogen-associated molecular patterns (PAMPs). CORE is a leucine-rich repeat receptor-like kinase with 22 LRRs, there additionally is a 6 amino acid insert at LRR 11. It consists of an extracellular domain that perceives an epitope, csp22, from the highly conserved nucleic acid binding motif RNP-1 of bacterial cold-shock proteins (CSPs), which are highly abundant proteins found in the cytosol of bacteria. Further domains are a single pass transmembrane domain and an intracellular kinase domain (The sequence encoding the kinase domain is not in this part). Interaction of CORE with brassinosteroid-associated kinase (BAK)1 is necessary for inducing an immune response in the plant. The dimerization of CORE and BAK1 depends on the csp22, the ligand of CORE. The function of CORE in S. lycopersicum has been confirmed by expressing the receptor in A. thaliana, which made the plant responsive to csp22, a PAMP that is otherwise not perceived by PRRs from A. thaliana.

Usage with mCherry

In this case, the C-terminal domain of CORE, entailing the intracellular kinase domain, was removed from the sequence. Instead, the C-terminal domain of the split mCherry was fused to the C-terminal domain via a 15 amino acid linker.

This part is used in the same way as Part:BBa_K3610048, meaning as a way to visualize the presence of the csp22 protein together with the coreceptor. It is, however, modified for expression in C. reinhardtii instead of S. cerevisiae.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 93
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 148
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1071
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1917