Difference between revisions of "Part:BBa K3989010"

Revision as of 16:29, 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 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 outer membrane vesicle (Figure 1).

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

Figure 2.

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]

@@ Line 5: / Line 5: @@
 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.
-To prove that ClyA can display certain plant immune elicitor to the surface of OMVs, we extracted OMVs and did western blot using anti-FLAG antibody.
+By doing western blot using anti-FLAG antibody, we proved that ClyA-elicitor fusion proteins indeed exist in outer membrane vesicle (Figure 1).
 [[File:ClyA_in_omp8_OMV.jpeg|500px]]