Coding

Part:BBa_K3610002

Designed by: Jonas Sebastian Trottmann   Group: iGEM20_UZurich   (2020-09-01)
Revision as of 18:17, 30 September 2020 by Jtrott (Talk | contribs)


BAK1 from Arabidopsis thaliana without signal sequence

This is a modification of the part Part:BBa_K3610001, which entails the sequence for the plant cell surface receptor BAK1 from A. thaliana. This modificated part does not contain the N-terminal signal sequence necessary for the protein to localize at the cell surface in A. thaliana. This allows the insertion of any signal sequence from other organisms. This part is thus better suited for the expression in organisms that do not recognize the original signal peptide.

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 our project, we used this receptor in the same manner as Part:BBa_K3610001. We expressed it in S. cerevisiae with a marker to observe expression and localization of the protein in the cell. For characterization see Part:BBa_K3610031.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1485
    Illegal PstI site found at 575
    Illegal PstI site found at 620
    Illegal PstI site found at 910
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1485
    Illegal PstI site found at 575
    Illegal PstI site found at 620
    Illegal PstI site found at 910
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1485
    Illegal PstI site found at 575
    Illegal PstI site found at 620
    Illegal PstI site found at 910
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1485
    Illegal PstI site found at 575
    Illegal PstI site found at 620
    Illegal PstI site found at 910
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None