Difference between revisions of "Part:BBa K3989010"

Revision as of 17:53, 16 October 2021

plant immune elicitor flg22 fused with ClyA for OMV display

In this construct, outer membrane-associated protein ClyA (Part:BBa_K811000) is used to display the plant immune elicitor flg22 (Part:BBa_K3286138) 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).

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

The immunogenicity of engineered OMVs was tested by Seedling Growth Inhibition (SGI) assay (Figure 2 & 3）, in which lower seedling weight indicates that higher immune responses are triggered (due to the trade-off between immunity and growth). Two OMV-producing E. coli strains were used, omp8 and tolB. In both strain, ClyA-flg22 and ClyA-elf18 can trigger stronger immune responses in A. thaliana col-0.

Figure 2. engineered E. coli omp8 OMVs can trigger stronger immune responses.

Figure 3. engineered E. coli tolB OMVs can trigger stronger immune responses.

Sequence and Features