Difference between revisions of "Part:BBa K3989010"
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<partinfo>BBa_K3989010 short</partinfo>
 
<partinfo>BBa_K3989010 short</partinfo>
  
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In this construct, outer membrane-associated protein ClyA is used to display the plant immune elicitor [[Part:BBa_K3286138|flg22]] to trigger plant immune responses. To make flg22 better recognized by plant immune receptor FLS2, we put mutiple linkers and a 3×FLAG tag between ClyA and flg22 to minimize the steric hindrance.
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In this construct, outer membrane-associated protein ClyA is used to display the plant immune elicitor [[Part:BBa_K3286138|flg22]] on the surface of bacterial outermembrane vesicles (OMVs) to trigger plant immune responses. To make flg22 better recognized by plant immune receptor FLS2, we put mutiple linkers and a 3×FLAG tag between ClyA and flg22 to minimize the steric hindrance.
  
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By doing western blot using anti-FLAG antibody, we proved that ClyA-elicitor fusion proteins indeed exist in outer membrane vesicle (Figure 1).
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By doing western blot using anti-FLAG antibody, we proved that ClyA-elicitor fusion proteins indeed exist in OMVs (Figure 1).
  
 
[[File:ClyA_in_omp8_OMV.jpeg|500px]]
 
[[File:ClyA_in_omp8_OMV.jpeg|500px]]
  
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Figure 1. ClyA protein with or without different PAMPs exists in outer membrane vesicles of E. coli omp8 strain.
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Figure 1. ClyA protein with or without elicitor fused exists in outer membrane vesicles of E. coli omp8 strain.
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[[File:omp8 OMVs on col-0.jpeg|500px]]
 
[[File:omp8 OMVs on col-0.jpeg|500px]]

Revision as of 17:36, 16 October 2021


plant immune elicitor flg22 fused with ClyA for OMV display

In this construct, outer membrane-associated protein ClyA is used to display the plant immune elicitor flg22 on the surface of bacterial outermembrane vesicles (OMVs) to trigger plant immune responses. To make flg22 better recognized by plant immune receptor FLS2, we put mutiple linkers and a 3×FLAG tag between ClyA and flg22 to minimize the steric hindrance.

By doing western blot using anti-FLAG antibody, we proved that ClyA-elicitor fusion proteins indeed exist in OMVs (Figure 1).

ClyA in omp8 OMV.jpeg

Figure 1. ClyA protein with or without elicitor fused exists in outer membrane vesicles of E. coli omp8 strain.


Omp8 OMVs on col-0.jpeg

Figure 2.

TolB OMVs on col-0.jpeg

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1105
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1105
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1105
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1105
    Illegal NgoMIV site found at 1099
    Illegal AgeI site found at 1063
  • 1000
    COMPATIBLE WITH RFC[1000]