Difference between revisions of "Part:BBa K2278001:Design"

 
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<partinfo>BBa_K2278001 short</partinfo>
 
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<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2278001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2278001 SequenceAndFeatures</partinfo>
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This DNA biobrick was designed in order to produce C8-CAI-1 of <i>Vibrio harveyi</i>  in <i>E. coli</i> strain.
  
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<p>The production of the <i>Vibrio harveyi</i> quorum sensing inducer (C8-CAI-1) is under the control of the cqsA gene coding for the CqsA synthase.
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<p> The BBa_K2278001 cames from the sensing module of the Croc’n cholera project <a href="http://2017.igem.org/Team:INSA-UPS_France">(team INSA-UPS-France 2017)</a>
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It was designed to produce C8-CAI-1 to simulate the presence of <i>Vibrio cholerae</i> in a water sample and so, to allow the validation of our <i>Vibrio cholerae</i> quorum sensing based detection system. </p>
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<p>The part includes <i>Vibrio harveyi</i> cqsA synthase under the control of an IPTG inducible promoter.
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The C8-CAI-1 producing system is inducible in order to avoid toxicity problems and high metabolic activity during cells growth. </p>
  
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<p><b>As the cqsA gene comes from <i>Vibrio harveyi</i> which is a BSL1 organism, it can be used as substitute in experiment about the <i>Vibrio cholerae</i> quorum sensing in BSL1 condition.</b></p>
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===Design Notes===
 
===Design Notes===
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The terminator  (BBa_B1006) is mutated substitution at the 35th pair (initial A->T) due to IDT complexity requirements for gBlocks  synthesis. The effect of the mutation was not investigated.
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<p>The part includes <i>Vibrio harveyi</i> cqsA synthase under the control of an IPTG inducible promoter.  
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The C8-CAI-1 producing system is inducible in order to avoid toxicity problems and high metabolic activity during cells growth. </p>
  
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Sequencing revealed that the VhCqsA construction slightly differs from the initial design, with a loss of the 9 last amino acid of the protein (position 382 to 391).
  
 
===Source===
 
===Source===

Revision as of 04:06, 16 October 2017

Vibrio harveyi C8-CAI-1 (quorum sensing inducer) generator

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 136
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

This DNA biobrick was designed in order to produce C8-CAI-1 of Vibrio harveyi in E. coli strain.

The production of the Vibrio harveyi quorum sensing inducer (C8-CAI-1) is under the control of the cqsA gene coding for the CqsA synthase.

The BBa_K2278001 cames from the sensing module of the Croc’n cholera project (team INSA-UPS-France 2017) It was designed to produce C8-CAI-1 to simulate the presence of Vibrio cholerae in a water sample and so, to allow the validation of our Vibrio cholerae quorum sensing based detection system.

The part includes Vibrio harveyi cqsA synthase under the control of an IPTG inducible promoter. The C8-CAI-1 producing system is inducible in order to avoid toxicity problems and high metabolic activity during cells growth.

As the cqsA gene comes from Vibrio harveyi which is a BSL1 organism, it can be used as substitute in experiment about the Vibrio cholerae quorum sensing in BSL1 condition.

Design Notes

The part includes Vibrio harveyi cqsA synthase under the control of an IPTG inducible promoter. The C8-CAI-1 producing system is inducible in order to avoid toxicity problems and high metabolic activity during cells growth.

Sequencing revealed that the VhCqsA construction slightly differs from the initial design, with a loss of the 9 last amino acid of the protein (position 382 to 391).

Source

This enzyme cames from Vibrio harveyi DNA genomic sequence.

Henke, J. and Bassler, B. (2004). Three Parallel Quorum-Sensing Systems Regulate Gene Expression in Vibrio harveyi. Journal of Bacteriology,186(20), pp.6902-6914. Sequence is available at: https://www.ebi.ac.uk/ena/data/view/AAT86008&display=text


References