Part:BBa_K3447000
xynD, which produces xylanase
The cvaA and cvaB genes encode components of a dedicated secretion system required for extracellular secretion of colistin. The CvaB protein encoded by the cvaB gene is the central membrane transporter of the bacteriocin 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.
- https://www.ncbi.nlm.nih.gov/nuccore/NC_007675.1?report=genbank&from=155164&to=156075
- https://www.ncbi.nlm.nih.gov/nuccore/NC_007675.1?report=genbank&from=156050&to=158164
We ordered the part DNA from a synthesis company.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.
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