Composite part: xylose operon BBa_K4358014
Short description: xylose operon with pqqC pqqD pqqE
Long description:
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 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.
Figure 1. It illustrated the mechanism of how xylose regulator protein interacts with xylose promoter and inhibits the performence of the downstream genes. The XylR would bind to the xylose binding site until the structure changed because of xylose.
Figure 2. xylose operon with pqqC pqqD pqqE (BBa_K4358014)
Composite part BBa_K4358013
Short Description: Multi-pqqA pET-28a
Long description:
Due to BBa_K3830012 used in 2021 NCHU_Taichung and the information displayed in wiki and registry, we knew that PqqA is a precursor for pyrroloquinoline quinone, PQQ, biosynthesis. Thus, the expression of pqqA is vital to the magnification of PQQ production. For the purpose of increasing the production of PQQ, we tried to double pqqA in our plasmid to magnify the expression of pqqA to enhance PQQ production.
Figure 1. Multi-pqqA pET28-A (BBa_K4358013)
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.
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.
Composite part BBa_K4358011
Short Description: Pchis PQQ plasmid.
Long description:
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.
Figure 1. Pchis PQQ plasmid (BBa_K4358011)
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.
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.
Composite part BBa_K4358016
Short Description: Pxyl with pqqA
Long description: We transformed this composite to BBa_K3830012 to amplify the synthesis of PqqA.
Figure 1. Pxyl with pqqA BBa_K4358016
