Part:BBa_K3606003

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 FudanBBa_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.

<img src="https://2020.igem.org/wiki/images/9/92/T--Fudan--img_mcbabcd.svg" />

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 2297
  Illegal PstI site found at 2330
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 2297
  Illegal PstI site found at 2330
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 2297
  Illegal PstI site found at 2330
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 2297
  Illegal PstI site found at 2330
  Illegal NgoMIV site found at 1959
  Illegal AgeI site found at 2131
  
• 1000
  COMPATIBLE WITH RFC[1000]

Results:

We constructed the plasmid and successfully expressed the mcbABCD, here is the electrophoresis map. 图

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. Thses 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

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

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

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.