Part:BBa_K4905021

Ompf-IL10-His

This composite part is made with the basic parts Outer membrane protein F (OmpF) (BBa_K2302005), Interleukin 10 (IL-10) (BBa_K554004), and Polyhistadine-tag (His-tag) (BBa_K3630006). This forms the construct OmpF-IL10-His. The TU Eindhoven 2023 team used it as a therapeutic against Inflammatory Bowel disease (IBD).

IL-10 is important for regulating inflammation^[1]. It is immunosuppressive with effect on macrophages and dendritic cells. Animal studies show that mice with IL-10 deficiency develop severe colitis, but not when recombinant IL-10 is systemically applied. With the anti-inflammatory effect of IL-10, it can be used to treat IBD and other autoimmune disorders. However, an oral intake or injection with only IL-10 induces side effects. The options are also limited because of the acid sensitivity of IL-10. Genetically modified organisms are an alternative for delivering IL-10 at the right place of the inflammation. In our case with E.coli BL21 cells as carrier.

For the expression of IL-10 by E.coli, OmpF is added to the construct. This is a major outer membrane porin that can secrete gene products from E.coli. It causes the construct to form an artificial transporter such that IL-10 can cross the cytoplasmic membrane. On the other side of IL-10, we added a His-tag. This is used for affinity purification of the construct.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] Pöhlmann, C., Brandt, M., Mottok, D. S., Zschüttig, A., Campbell, J. W., Blattner, F. R., Frisch, D., & Gunzer, F. (2012). Periplasmic Delivery of Biologically Active Human Interleukin-10 in Escherichia coli via a Sec-Dependent Signal Peptide. Journal of Molecular Microbiology and Biotechnology, 22(1), 1–9. https://doi.org/10.1159/000336043