Part:BBa_K3989014
double cellulose binding domain fused with ClyA for OMV display
In this construct, outer membrane-associated protein ClyA (Part:BBa_K811000) is used to display the double cellulose-binding domain (dCBD, Part:BBa_K1321340) to the surface of bacterial outer membrane vesicles (OMVs). To make dCBD better displayed, we put multiple linkers and a 3×FLAG tag between ClyA and dCBD to minimize the steric hindrance.
Usage and Biology
Chracterization by team 2021 UZurich
Seedling growth inhibition (SGI) assay was performed to test whether displaying cellulose-binding domain on OMV surface can enhance the plant immune response (the lower seedling weight, the stronger plant immune response). OMV with only ClyA (blue boxes) was used as the negative control.
As the figure shown below, the seedling weight of negative control group gradually decreases as OMV concentration increases. However, for ClyA-dCBD, the seedling weight dramatically decreases between concentrations of 0 and 2.5 ug/mL, but doesn't decrease a lot as concentration further increases, which means the immune response might get saturated.
The reason might be that engineered OMVs with dCBD can get enriched by cellulose in cell wall, which makes OMVs more effective at low concentraion.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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