Difference between revisions of "Part:BBa K3447000"
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Colicin V is a non-ribosome-encoded antibacterial peptide produced by many <i>E. coli</i> strains and other members of the <i>Enterobacteriaceae</i> 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 <i>E. coli</i>.<br><br>
 
Colicin V is a non-ribosome-encoded antibacterial peptide produced by many <i>E. coli</i> strains and other members of the <i>Enterobacteriaceae</i> 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 <i>E. coli</i>.<br><br>
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The <i>cvaA</i> and <i>cvaB</i> genes encode components of a dedicated secretion system required for extracellular secretion of colicin V.
+
The <i>cvaA</i> and <i>cvaB</i> genes encode components of a dedicated secretion system required for extracellular secretion of colicin V.<br><br>
  
 
==<b>Design</b>==
 
==<b>Design</b>==
 
===Design Notes===
 
===Design Notes===
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There is a repetitive sequence between <i>cvaA</i> and <i>cvaB</i> gene. In order not to increase restriction sites, we modified the natural gene and added some nonsense mutations, and other than that there is no need to pay attention.
+
There is a repetitive sequence between <i>cvaA</i> and <i>cvaB</i> gene. In order not to increase restriction sites, we modified the natural gene and added some nonsense mutations, and other than that there is no need to pay attention.<br><br>
  
 
===Source===
 
===Source===

Revision as of 05:24, 5 September 2020

luxI, an autoinducer synthetase for AHL


The cvaA and cvaB genes encode components of a dedicated secretion system required for extracellular secretion of colicin V. The CvaB protein encoded by the cvaB gene is the central membrane transporter of the colicin V secretion system in Escherichia coli. CvaB, together with CvaA and TolC proteins, direct the secretion of colicin V through the cleavage of the bisglycine leader peptide. The sequence alignment shows that CvaB protein belongs to the ABC superfamily.


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 cvaA and cvaB genes encode components of a dedicated secretion system required for extracellular secretion of colicin V.

Design

Design Notes

There is a repetitive sequence between cvaA and cvaB gene. In order not to increase restriction sites, we modified the natural gene and added some nonsense mutations, and other than that there is no need to pay attention.

Source

We will use cvaAB to express channel proteins for the transport of ColV out of the cell membrane.This gene is derived from Escherichia coli (strain: A2363), we get the genomic sequence by NCBI.

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.