Part:BBa_K4161100
Csg surface display system on E. coli Nissle 1917
We intend to display VHH (in this example, JMK-H2) on the surface of E. coli Nissle 1917 through curli fibers and seliver it to intestines to treat shigella infection. The production of culi fibers is encoded by csg genes. The csgDEFG genes encodes proteins that secrete curli fiber units - csgA & csgB, while csgC protein prevents fiber formation in the bacterium. As shown in the plasmid, the VHH (JMK-H2) is conjugated with csgA protein by flexible linker FE36 containing E-tag, and will be transported to the extracellular space, then displayed along with csgA protein. Apart from the E tag, the VHH is also marked with 3xFlag tag at its C terminal. pUC19 is chosen as the plasmid backbone, and the pTlpA (temperature responsive promotor) was added to ensure normal expression of csg genes in gut environment.
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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