Difference between revisions of "Part:BBa K4239010"

 
 
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<p><i>axe/txe</i> is a toxin/antitoxin system to be used on the same plasmid than an interest gene (the <i>fiatlux</i> operon, in our project). This method of selection ensures that all the bacteria present on the plant have the plasmid, and can hence emit light. Toxins have a longer life span in cells than antitoxins, so if the bacteria lose their plasmid, the toxin is no longer destroyed by the antitoxin. This results in the death of the bacteria, thus assuring the death of all <i>fiatlux</i> non-carriers. As a consequence, by involving a selective pressure, the toxin/antitoxin system increases the stability of the plasmid in the bacteria. </p>
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<h2>Sequence and Features</h2>
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<h2>Construction</h2>
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<p>The gene <i>axe/txe</i> was already available in a plasmid pUC-GFP-<i>axe/txe</i>. A PCR was performed on the plasmid to get it as a fragment</p>
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<h2>References</h2>
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<p><i> Gregor C, Gwosch KC, Sahl SJ, Hell SW. Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging. Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):962-967. doi: 10.1073/pnas.1715946115. Epub 2018 Jan 16. PMID: 29339494; PMCID: PMC5798359.</i> </p>
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Latest revision as of 15:36, 12 October 2022


toxin/antitoxin gene axe/txe


Description

axe/txe is a toxin/antitoxin system to be used on the same plasmid than an interest gene (the fiatlux operon, in our project). This method of selection ensures that all the bacteria present on the plant have the plasmid, and can hence emit light. Toxins have a longer life span in cells than antitoxins, so if the bacteria lose their plasmid, the toxin is no longer destroyed by the antitoxin. This results in the death of the bacteria, thus assuring the death of all fiatlux non-carriers. As a consequence, by involving a selective pressure, the toxin/antitoxin system increases the stability of the plasmid in the bacteria.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 167
    Illegal PstI site found at 1631
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 167
    Illegal PstI site found at 1631
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1343
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 167
    Illegal PstI site found at 1631
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 167
    Illegal PstI site found at 1631
  • 1000
    COMPATIBLE WITH RFC[1000]


Construction

The gene axe/txe was already available in a plasmid pUC-GFP-axe/txe. A PCR was performed on the plasmid to get it as a fragment

References

Gregor C, Gwosch KC, Sahl SJ, Hell SW. Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging. Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):962-967. doi: 10.1073/pnas.1715946115. Epub 2018 Jan 16. PMID: 29339494; PMCID: PMC5798359.