Composite

Part:BBa_K3606086

Designed by: Yuanyuan Liu   Group: iGEM20_Fudan   (2020-10-23)


PlacIq_lacI_Ptac driven mcbABCD

This is a plamid with high expression level.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.

T--Fudan--img_mcbabcd.png

Figure1. structure of mcbABCD

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2326
    Illegal PstI site found at 2359
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2326
    Illegal PstI site found at 2359
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2326
    Illegal PstI site found at 2359
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2326
    Illegal PstI site found at 2359
    Illegal NgoMIV site found at 1988
    Illegal AgeI site found at 2160
  • 1000
    COMPATIBLE 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.

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