Part:BBa_K2817002
IL10-flag
Interleukin-10 is a natural immunosuppressive cytokine that is normally released by regulatory T cells and plays an important role in intestinal homeostasis. In our engineered bacteria, we hope to induce the release of interleukin-10 through the inflammatory signal nitric oxide to achieve drug release controllability.
Usage and Biology
We inserted this part into plasmid pCDFDuet-1. So the expression of this part can be induced by IPTG. We transformed the human IL10-Flag plasmid into BL21, incubated at 37 ℃ overnight and then diluted to OD600 = 0.2. After 2 h of growth at 37 ℃, IPTG was added to induce IL10 expression at 30 ℃ for 16 h. Then, the bacterial solution was lysed and the expressed IL10 was detected by Western Blot (Figure 1). From this result, IL10 was successfully expressed in our constructed plasmid. As a widely validated and used anti-inflammatory interleukin, IL10 has been identified with critical functions in immunosuppression in human [1].
Figure 1. Western blot analyses using a flag antibody on bacterial lysate to detect IL10. Lane 1: Negative control (cell lysate without IPTG induction); Lane 2: 0.5mM IPTG, Lane3: 1mM IPTG induction for 16h at 30℃.
[1] Steidler, L. Treatment of murine colitis by Lactococcus lactis secreting interleukin-10. Science 289, 1352–1355 (2000).
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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