Difference between revisions of "Part:BBa K3606003"
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We have successfully expressed mcbA and mcbC for the foundamental validation of the antimicrobial peptide(mccb17) expression. | We have successfully expressed mcbA and mcbC for the foundamental validation of the antimicrobial peptide(mccb17) expression. | ||
https://2020.igem.org/wiki/images/f/f3/T--Fudan--Poster_Mcb-gels.jpg | https://2020.igem.org/wiki/images/f/f3/T--Fudan--Poster_Mcb-gels.jpg | ||
− | Figure2. gel map for mcbA | + | Figure2. gel map for mcbA, mcbC, mcbE and mcbF |
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Revision as of 02:23, 5 December 2020
mcbABCD
This part produces an antibiotic originated from Escherichia coli that targets a bacterial topoisomerase, DNA gyrase.
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 an antibiotic coding gene cluster with leader peptide and proteins for its maturation.
Figure1. structure of mcbABCD
Results
We have successfully expressed mcbA and mcbC for the foundamental validation of the antimicrobial peptide(mccb17) expression. Figure2. gel map for mcbA, mcbC, mcbE and mcbF
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 2297
Illegal PstI site found at 2330 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 2297
Illegal PstI site found at 2330 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 2297
Illegal PstI site found at 2330 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 2297
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Illegal AgeI site found at 2131 - 1000COMPATIBLE WITH RFC[1000]
Further Application
For futher application, this part is provided as an antimicrobial peptide(mccb17) expressing example and are effective for a wide range of microbes. This part can be used together with BBa_K3606004 to provide immunity for the engineered strain and create survival advantage. 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.