Part:BBa_K3447002

xynD, which produces xylanase

The cvaC gene produces colicin V, which is also called ColV, is a type of bacteriocin secreted by E. coli. Besides, it has a killing effect on other closely related bacteria that cannot secrete specific colicin immunity protein, which is synthesized by cvi gene.

Usage and Biology

Colicin V is a non-ribosome-encoded antibacterial peptide produced by many E. coli strains and other members of the Enterobacteriaceae family, which kills bacteria by destroying the membrane potential of sensitive target cells. Due to the existence and composition of the outer membrane of Gram-negative bacteria, natural colicin V is usually inactive to it, but its expression in lactic acid bacteria has also been proposed in the literature: naturally generated colicin V can destroy sensitive targets cell membrane potential to kill Gram-positive bacteria. The gene encoding colicin V is derived from the pAPEC-O2-ColV plasmid of E. coli.

The cvaC gene encodes a 103 amino acids' precursor of ColV (preColV), and cvi encodes a protein of 78 amino acids, which can provide immunity against the bactericidal activity of ColV. Therefore, only when the cvaC structural gene and the cvi (237 bp) immune gene exist at the same time, the E. coli itself will not be killed.

We will use cvaC-cvi to express ColV and immunity protein, thereby killing "enemy" bacteria.

Design

Design Notes

There is a repetitive sequence between cvi and cvaC gene, and other than that there is no need to pay attention.

Source

This gene is derived from Escherichia coli (strain: A2363), we get the genomic sequence by NCBI.

• https://www.ncbi.nlm.nih.gov/nuccore/NZ_CP033091.2?report=genbank&from=43597&to=43908&strand=true
  
• https://www.ncbi.nlm.nih.gov/nuccore/NZ_CP033091.2?report=genbank&from=43886&to=44122&strand=true
  

We ordered the part DNA from a synthesis company.

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.rc site found at 712
  

References

1. Coproduction of colicin V and lactic acid bacteria bacteriocins in lactococci and enterococci strains of biotechnological interest., Journal of Applied Microbiology, 122, 1159-1167.
2. Colicin V can be produced by lactic acid bacteria.(1999), Letters in Applied Microbiology, 29, 37-41.
3. Mutational Analysis of Cvab, an ABC Transporter Involved in the Secretion of Active Colicin V., Kai-Hui Wu., Ying-Hsin Hsieh., Phang C. Tai, Plos One, 7(4), e35382.
4. Purification and Characterization of Colicin V from Escherichia coli Culture Supernatants.(1994), Biochemistry, 33, 6911-6917.