Part:BBa_K3788015

Modified lysis device

Introduction :

This part contains the modified lysis device. It is composed of a fragment of the cal gene encoding the CaL lysis protein. It also contains the caa and cai genes coding respectively for the ColA toxin (colicin A) and the CaI immunity protein.

In nature, the caa and cai genes are in operon, and the gene encoding the immunity protein is constitutively expressed and is found on the antisense strand of the operon. The regulation of this system is complex and contains many regulatory elements.

This system is located on a plasmid present in certain populations of wild E. coli. It induces the accumulation of ColA toxin in the host cell and then causes the latter to die by the action of the lysis protein. This releases ColA into the medium and kills cells that do not have the immunity gene present on this plasmid.

The cal gene encodes the CaL protein, a 28 to 41 residue lipopeptide, which is produced as a precursor and then processed. When mature, it forms pores in the membrane, causing the host to near lysis and the leakage of cellular components into the medium.

Design :

The presence of the truncated caa and cai genes allows the characterization of these different elements.

- The presence of the caa fragment allows the addition at 5 ′ of a gene encoding for GFP (BBa_E0040) and thus allows the conservation of regulatory elements internal to the nucleotide sequence of caa.

- The cai fragment allows the addition at 3′ of a gene encoding for RFP (BBa_E1010).

These fusion genes allow the construction of the composite parts BBa_K3788021 and K3788020 under the control of the lexA-repressed promoter (BBa_K3788017).

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 585
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 905
  Illegal BamHI site found at 578
  Illegal XhoI site found at 454
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 654
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 408