Part:BBa_K3165014

ccdB (L83S)

The CcdB protein derived from the ccdAB operon in Eschereschia coli is subjected to a core mutation at the 83rd amino acid (changing Leucine to Serine) resulting in instability of the protein rendering it less potent for the cell.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 216
  

Biology

Controller of Cell Division or Death B (CcdB) is the toxic component of the Eschereschia coli CcdAB anti-toxin toxin system. It is a globular, dimeric protein with 101 residues per protomer, involved in the maintenance of F plasmid in cells by a mechanism involving its binding and the poisoning of DNA Gyrase A which leads to the breaking of the Double-stranded DNA in the bacteria and eventually death. The L83S mutant of the wild type CcdB protein is being mutated at the 83^th amino acid residue position from Leucine to Serine in the core region of the protein.

Usage

This coding sequence for CcdB L83S mutant is used with araBp (BBa_K3165015) upstream to regulate the production of the protein of interest only in the presence of arabinose and addition of glucose stops the leaky transcription. The generator of the ccdB L83S is (BBa_K31650148)

References

1. Anusmita Sahoo, Shruti Khare, Sivasankar Devanarayanan, Pankaj C. Jain, and Raghavan Varadarajan Residue proximity information and protein model discrimination using saturation-suppressor mutagenesis doi: 10.7554/eLife.09532