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| − | We used McbA-D to inhibit the growth of other bacteria in human intestine so as to enhance Nissle's competitiveness,as well as to reduce the risk of illness caused by some opportunistic pathogens. We introduced McbE-G to inhibit the growth of other bacteria in human intestine so as to enhance Nissle's competitiveness,as well as to reduce risk of illness caused by some opportunistic pathogens.At the same time, we put McbE-G under the control of pTetR in order to response to AHL-LuxR.
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| − | <h2>Background:</h2>
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| − | <h2>Design:</h2>
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| − | Here, we tried to improve the former antimicrobial peptide(mccb17) expressing system of 2019 Fudan [[BBa_K3245010]]. By dividing into the peptide expressing parts and the immunity parts, we wanted to manipulate their expression level with more efficiency.
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| − | https://2020.igem.org/wiki/images/b/b4/T--Fudan--img_mcbabcd.png
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| − | Figure. mcbABCD
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| − | https://2020.igem.org/wiki/images/7/74/T--Fudan--img_mcbefg.png
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| − | Figure. mcbEFG
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| − | <h2>Usage and Biology:</h2>
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| − | We introduced mcbABCD to our improved antibiotic expressing system to inhibit the growth of other bacteria in human intestine so as to enhance Nissle's competitiveness, as well as to reduce risk of illness caused by some opportunistic pathogens.Driven by a series of constitutive promoters with different strength which include P12, mcbABCD is expressed in E. coli in a controlled, reliable manner.
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| − | McbEFG works as an adjusting part in quorum sensing system. When the number of engineered bacteria is low, the part will help export antibiotic and provide self immunity in order to help proliferate. PtetR determines the expression of mcbEFG when the number of bacteria is at different level.
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| − | <span class='h3bb'>Sequence and Features</span>
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| | <partinfo>BBa_K3606068 SequenceAndFeatures</partinfo> | | <partinfo>BBa_K3606068 SequenceAndFeatures</partinfo> |
| − | <h2>Results:</h2>
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| − | We constructed the plasmid and successfully expressed the system, here is the electrophoresis map.
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| − | 图
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| − | <h2>Further Application:</h2>
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| − | For futher application, this part expresses antimicrobial peptide(mccb17) as well as providing immunity to create survival advantage for the engineered strain. These part are especially useful to be expressed in vivo because research has proved that it can also ease the inflammation in intestine by limiting the expansion of related pathogens and pathobionts.
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| − | <h2>References:</h2>
| + | ===Design Notes=== |
| − | Collin F, Maxwell A. The Microbial Toxin Microcin B17: Prospects for the Development of New Antibacterial Agents. J Mol Biol. 2019;431(18):3400–3426. doi:10.1016/j.jmb.2019.05.050
| + | McbABCD and McbEFG are placed in reverse, and each is independently expressed under different promoters with their own terminator. |
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| − | S. Duquesne, D. Destoumieux-Garzón, J. Peduzzi, S. Rebuffat. Microcins, gene-encoded antibacterial peptides from enterobacteria
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| − | Sassone-Corsi M, Nuccio SP, Liu H, Hernandez D, Vu CT, Takahashi AA, Edwards RA, Raffatellu M. Microcins mediate competition among Enterobacteriaceae in the inflamed gut. Nature. 2016 Dec 8;540(7632):280-283.
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| − | Mutalik VK, Guimaraes JC, Cambray G, Lam C, Christoffersen MJ, Mai QA, Tran AB, Paull M, Keasling JD, Arkin AP, Endy D. Precise and reliable gene expression via standard transcription and translation initiation elements. Nat Methods. 2013 Apr;10(4):354-60. doi: 10.1038/nmeth.2404. Epub 2013 Mar 10. PMID: 23474465.
| + | ===Source=== |
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| | + | McbABCD and McbEFG orginally come from a plasmid of Escherichia coli. |
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| − | <!-- Uncomment this to enable Functional Parameter display
| + | ===References=== |
| − | ===Functional Parameters=== | + | |
| − | <partinfo>BBa_K3606029 parameters</partinfo>
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McbABCD and McbEFG are placed in reverse, and each is independently expressed under different promoters with their own terminator.
