Part:BBa_K4633008

Colicin D - Bacteriocin from E. coli

Usage and Biology

Colicins are a class of bacteriocins that Escherichia coli produces to eliminate competing microorganisms. One of these colicins, known as Colicin D, is a substantial protein weighing 75 kDa, and it poses no harm to humans ​[1]. ​It consists of three distinct segments: a translocation domain, a receptor-binding domain, and a cytotoxic domain ​[2].​ It is worth noting that the cytotoxic domain is responsible for the specific cleavage of the anticodon loop in all four tRNA-Arg isoacceptors. This action disrupts the critical process of protein synthesis, ultimately leading to cellular demise ​[2]​.

A total of 697 amino acids make up the Colicin D segment ​[3].​ It was intended to be utilized and tested in Bacillus subtilis 168, a Gram-positive model organism chosen for our project. It was intended later to be expressed in Lactobacillus crispatus.

Additional information can be found on our wiki.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1273
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1524
  Illegal AgeI site found at 506
  Illegal AgeI site found at 1470
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1725
  

References

• [1] X. Jin, W. Kightlinger, Y. C. Kwon, and S. H. Hong, “Rapid production and characterization of antimicrobial colicins using Escherichia coli-based cell-free protein synthesis,” Synth Biol, vol. 3, no. 1, Jan. 2018, doi: 10.1093/SYNBIO/YSY004.
• [2] M. Graille, L. Mora, R. H. Buckingham, H. Van Tilbeurgh, and M. De Zamaroczy, “Structural inhibition of the colicin D tRNase by the tRNA-mimicking immunity protein,” EMBO Journal, vol. 23, no. 7, pp. 1474–1482, Apr. 2004, doi: 10.1038/SJ.EMBOJ.7600162.
• [3] U. Roos, R. E. Harkness, and V. Braun, “Assembly of colicin genes from a few DNA fragments. Nucleotide sequence of colicin D,” Mol Microbiol, vol. 3, no. 7, pp. 891–902, 1989, doi: 10.1111/J.1365-2958.1989.TB00238.X.