Difference between revisions of "Part:BBa K3924033"
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==Usage and Biology== | ==Usage and Biology== | ||
| − | We bioengineered Lactococcus lactis to express the Listeria adhesion protein (LAP) from a non-pathogenic Listeria (L. innocua). L. lactis colonize the intestine, reduce Lm mucosal colonization and systemic dissemination, and protect mice from lethal infection. L. lactis expressing LAP has been reported to competitively exclude Listeria(Lm) by occupying the surface presented LAP receptor, and can ameliorate the Lm-induced intestinal barrier dysfunction by blocking the nuclear factor-κB and myosin light chain kinase-mediated redistribution of the major epithelial junctional proteins. <sup>[1]</sup> | + | We bioengineered <i>Lactococcus lactis</i> to express the Listeria adhesion protein (LAP) from a non-pathogenic Listeria (<i>L. innocua</i>). <i>L. lactis</i> colonize the intestine, reduce Lm mucosal colonization and systemic dissemination, and protect mice from lethal infection. <i>L. lactis</i> expressing LAP has been reported to competitively exclude Listeria(Lm) by occupying the surface presented LAP receptor, and can ameliorate the Lm-induced intestinal barrier dysfunction by blocking the nuclear factor-κB and myosin light chain kinase-mediated redistribution of the major epithelial junctional proteins. <sup>[1]</sup> |
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==Design and Construction== | ==Design and Construction== | ||
| − | As our engineered probiotics are to be used in IBD treatment as prescribed drugs instead of dietary supplements, we deem it unnecessary to incorporate conditional promoters into our design. We therefore use the constitutive expression plasmid pMG36e as the backbone. Also, being a transfer plasmid, pMG36e can replicate in both E.coli and L.lactis, which saved us a lot of trouble in transformation.<br/> | + | As our engineered probiotics are to be used in IBD treatment as prescribed drugs instead of dietary supplements, we deem it unnecessary to incorporate conditional promoters into our design. We therefore use the constitutive expression plasmid pMG36e as the backbone. Also, being a transfer plasmid, pMG36e can replicate in both <i>E.coli</i> and <i>L.lactis</i>, which saved us a lot of trouble in transformation.<br/> |
| − | [[T--Tsinghua--LAP_plasmid.png|center|600px|thumb|'''Figure 1: The design of LAP''']] | + | [[Image: T--Tsinghua--LAP_plasmid.png|center|600px|thumb|'''Figure 1: The design of LAP''']] |
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==Reference== | ==Reference== | ||
[1] Drolia, R., Amalaradjou, M. A. R., Ryan, V., Tenguria, S., Liu, D., Bai, X., ... & Bhunia, A. K. (2020). Receptor-targeted engineered probiotics mitigate lethal Listeria infection. Nature communications, 11(1), 1-23.<br/> | [1] Drolia, R., Amalaradjou, M. A. R., Ryan, V., Tenguria, S., Liu, D., Bai, X., ... & Bhunia, A. K. (2020). Receptor-targeted engineered probiotics mitigate lethal Listeria infection. Nature communications, 11(1), 1-23.<br/> | ||
Latest revision as of 17:47, 21 October 2021
LAP
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]
Profile
Name: LAP
Base Pairs: 2925bp
Origin: Listeria innocua, synthetic
Properties: Listeria adhesion protein (LAP) from a non-pathogenic Listeria (L. innocua)
Usage and Biology
We bioengineered Lactococcus lactis to express the Listeria adhesion protein (LAP) from a non-pathogenic Listeria (L. innocua). L. lactis colonize the intestine, reduce Lm mucosal colonization and systemic dissemination, and protect mice from lethal infection. L. lactis expressing LAP has been reported to competitively exclude Listeria(Lm) by occupying the surface presented LAP receptor, and can ameliorate the Lm-induced intestinal barrier dysfunction by blocking the nuclear factor-κB and myosin light chain kinase-mediated redistribution of the major epithelial junctional proteins. [1]
Design and Construction
As our engineered probiotics are to be used in IBD treatment as prescribed drugs instead of dietary supplements, we deem it unnecessary to incorporate conditional promoters into our design. We therefore use the constitutive expression plasmid pMG36e as the backbone. Also, being a transfer plasmid, pMG36e can replicate in both E.coli and L.lactis, which saved us a lot of trouble in transformation.
Reference
[1] Drolia, R., Amalaradjou, M. A. R., Ryan, V., Tenguria, S., Liu, D., Bai, X., ... & Bhunia, A. K. (2020). Receptor-targeted engineered probiotics mitigate lethal Listeria infection. Nature communications, 11(1), 1-23.