Difference between revisions of "Part:BBa K3606004"
 
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[[File: T--Fudan--img_mcbefg.png|none|600px|thumb]]
 
[[File: T--Fudan--img_mcbefg.png|none|600px|thumb]]
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Figure1. structure of mcbEFG
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<h2>Results:</h2>
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We have successfully expressed mcbE and mcbF for the fundamental validation of the antimicrobial peptide(mccb17) expression.
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We've constructed the plasmid for mcbG expression, yet the results of colony PCR is false positive thus further validation is needed.
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https://2020.igem.org/wiki/images/f/f3/T--Fudan--Poster_Mcb-gels.jpg
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Figure2. gel map for mcbA, mcbC, mcbE and mcbF
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<h2>Usage and Biology:</h2>
 
<h2>Usage and Biology:</h2>

Latest revision as of 02:35, 5 December 2020


mcbEFG

This part 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.

Background:

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 firstly test whether the polycistron could work properly and separately, then manipulate their expression level with more efficiency.

Design:

This part is a immunity system coding gene cluster with efflux and immunity related protein.

T--Fudan--img mcbefg.png

Figure1. structure of mcbEFG

Results:

We have successfully expressed mcbE and mcbF for the fundamental validation of the antimicrobial peptide(mccb17) expression. We've constructed the plasmid for mcbG expression, yet the results of colony PCR is false positive thus further validation is needed. T--Fudan--Poster_Mcb-gels.jpg Figure2. gel map for mcbA, mcbC, mcbE and mcbF


Usage and Biology:

This part 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.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Further Application:

For futher application, this part is provided as an immunity system for antimicrobial peptide(mccb17). This part can be used together with BBa_K3606003 to express antimicrobial peptide(mccb17) and 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.

Reference

[1] 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

[2] S. Duquesne, D. Destoumieux-Garzón, J. Peduzzi, S. Rebuffat. Microcins, gene-encoded antibacterial peptides from enterobacteria

[3] 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.