Plasmid

Part:BBa_K4358011

Designed by: HUNG. CHIEN-CHUN   Group: iGEM22_NCHU_Taichung   (2022-09-16)


Pchis PQQ Plasmid

Gene expression with xylose operon can be induced by the addition of xylose. When the inducer, xylose, is presented, xylose would bind to the xylose regulator protein, XylR, and change its structure. Hence, xylR wouldn’t bind to the operator(XylR-binding site) to inhibit the downstream gene of the xylR-binding site.

So we design the plasmid with Pxyl, Xylr and XylR-binding site for the pqqC, pqqD and pqqE Then we transformed pqqC, pqqD and pqqE in the downstream of the xylR-binding site in addition to controlling the expression of pqqC, pqqD and pqqE. If PqqC, PqqD and PqqE were required to be expressed , the xylose was present in the culture surrounding; if not, there was no PqqC, PqqD and PqqE.


T--NCHU_Taichung--NCHU_2022_Figure_Q.png

Figure 1. Pchis PQQ plasmid (BBa_K4358011)



T--NCHU_Taichung--NCHU_2022_Figure_B.png

Figure 2. Wound healing assay(Normal HaCat cell) The HaCat cell was used to mimic wounds and the growth of the cells was observed to simulate the wound-healing situation. There were three treatments, Natto extracts, pyrroloquinoline quinone (PQQ), and the mixture treatments conducted and showed better recovery than the control group after 24hr culturing.




T--NCHU_Taichung--NCHU_2022_Figure_C.png

Figure 3. Wound healing assay (Diabetes induced HaCat cell) The diabetic-like cells were induced by the medium supplemented with 25 mM glucose and 10 nM insulin and were treated with Natto extracts, pyrroloquinoline quinone(PQQ), and the mixture composed of both. Natto extracts, pyrroloquinoline quinone(PQQ), and the mixture treatments showed better recovery than the control group after 24hr culturing.



Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3751
    Illegal XbaI site found at 1070
    Illegal XbaI site found at 1094
    Illegal XbaI site found at 3002
    Illegal XbaI site found at 3842
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3751
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3751
    Illegal BglII site found at 4019
    Illegal XhoI site found at 786
    Illegal XhoI site found at 1386
    Illegal XhoI site found at 3719
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3751
    Illegal XbaI site found at 1070
    Illegal XbaI site found at 1094
    Illegal XbaI site found at 3002
    Illegal XbaI site found at 3842
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3751
    Illegal XbaI site found at 1070
    Illegal XbaI site found at 1094
    Illegal XbaI site found at 3002
    Illegal XbaI site found at 3842
    Illegal NgoMIV site found at 2870
    Illegal NgoMIV site found at 4011
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1998
    Illegal SapI site found at 1196


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Parameters
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