Difference between revisions of "Part:BBa K4905017"
 
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For the expression of IL-10 by <i>E.coli</i>, a signal sequence is needed for the efficient production with the Sec secretion pathway. This is why PelB is added to IL-10. On the other side of IL-10, we added a His-tag. This is used for affinity purification of the construct.  
 
For the expression of IL-10 by <i>E.coli</i>, a signal sequence is needed for the efficient production with the Sec secretion pathway. This is why PelB is added to IL-10. On the other side of IL-10, we added a His-tag. This is used for affinity purification of the construct.  
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<span class='h3bb'><h1>Sequence and Features</h1></span>
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<partinfo>BBa_K4905017 SequenceAndFeatures</partinfo>
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<h1>References</h1>
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[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
 
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===Usage and Biology===
 
===Usage and Biology===
  
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Latest revision as of 09:18, 26 September 2023


PelB-IL10- His

This composite part is made with the basic parts Pectate Lyase B (PelB) (BBa_K2302005), Interleukin 10 (IL-10) (BBa_K554004), and Polyhistadine-tag (His-tag) (BBa_K3630006). This forms the construct PelB-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, a signal sequence is needed for the efficient production with the Sec secretion pathway. This is why PelB is added to IL-10. 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