Difference between revisions of "Part:BBa K3447000"
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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 CvaB protein encoded by the <i>cvaB</i> gene is the central membrane transporter of the colicin V secretion system in <i>Escherichia coli</i>. 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.<br><br>
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LuxI is the synthetase of AHL from <i>Vibrio fischeri</i>.<br>
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===Usage and Biology===
 
===Usage and Biology===
  
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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>
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LuxI gene is an important gene for gram-negative bacteria to conduct quorum sensing communication. It is responsible for the synthesis of AHL signaling molecules. AHL, an acyl hyperserine lactone, is the main signal molecule for gram-negative bacteria to communicate quorum sensing information. <i>E. coli</i> is a typical Gram-negative bacterium, but its quoral-sensing system 1 is not complete. It lacks the luxI gene, so it cannot synthesize AHL synthase and thus AHL signal molecules. We hope to introduce the luxI gene into Escherichia coli to produce AHL signaling molecules. (AHL is a class of signaling molecules in which the luxI gene differs from the luxR gene in different species of bacteria. The binding of signal molecules to receptor proteins is species-specific which can prevent interference from other species. However, <i>E. coli</i> is an exception that can sense all kinds of AHL signaling molecules).<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.<br><br>
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The constituent produces AHL: <partinfo>BBa_K3447108</partinfo><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.<br><br>
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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===
  
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We will use cvaAB to express channel proteins for the transport of ColV out of the cell membrane.This gene is derived from <i>Escherichia coli</i> (strain: A2363), we get the genomic sequence by NCBI.<br>
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We get the sequence data on GenBank and we ordered the part DNA from a synthesis company.
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*https://www.ncbi.nlm.nih.gov/nuccore/NC_007675.1?report=genbank&from=155164&to=156075<br>
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*https://www.ncbi.nlm.nih.gov/nuccore/NC_007675.1?report=genbank&from=156050&to=158164<br>
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We ordered the part DNA from a synthesis company.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3447002 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3447000 SequenceAndFeatures</partinfo>
  
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K3447002 parameters</partinfo>
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<partinfo>BBa_K3447000 parameters</partinfo>
 
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===References===
 
===References===
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1. Coproduction of colicin V and lactic acid bacteria bacteriocins in lactococci and enterococci strains of biotechnological interest., <i>Journal of Applied Microbiology</i>, <b>122</b>, 1159-1167.<br>
 
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2. Colicin V can be produced by lactic acid bacteria.(1999), <i>Letters in Applied Microbiology</i>, <b>29</b>, 37-41.<br>
 
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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, <i>Plos One</i>, <b>7(4)</b>, e35382.<br>
 
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4. Purification and Characterization of Colicin V from Escherichia coli Culture Supernatants.(1994), <i>Biochemistry</i>, <b>33</b>, 6911-6917.<br>
 

Latest revision as of 10:39, 27 October 2020

luxI, an autoinducer synthetase for AHL


LuxI is the synthetase of AHL from Vibrio fischeri.

Usage and Biology

LuxI gene is an important gene for gram-negative bacteria to conduct quorum sensing communication. It is responsible for the synthesis of AHL signaling molecules. AHL, an acyl hyperserine lactone, is the main signal molecule for gram-negative bacteria to communicate quorum sensing information. E. coli is a typical Gram-negative bacterium, but its quoral-sensing system 1 is not complete. It lacks the luxI gene, so it cannot synthesize AHL synthase and thus AHL signal molecules. We hope to introduce the luxI gene into Escherichia coli to produce AHL signaling molecules. (AHL is a class of signaling molecules in which the luxI gene differs from the luxR gene in different species of bacteria. The binding of signal molecules to receptor proteins is species-specific which can prevent interference from other species. However, E. coli is an exception that can sense all kinds of AHL signaling molecules).
The constituent produces AHL: BBa_K3447108

Design

Design Notes

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 get the sequence data on GenBank and 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
    COMPATIBLE WITH RFC[1000]



References