Difference between revisions of "Part:BBa K3962342"

 
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<partinfo>BBa_K3962342 short</partinfo>
 
<partinfo>BBa_K3962342 short</partinfo>
  
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This construct is composed of L-arabinose-mediate inducible promoter pBAD and toxin CcdB. CcdB/CcdA is a type II toxin/antitoxin system originated from E. coli. In our project, we used this construct to test the toxicity of toxin CcdB, which provided us with more insights to regulate the toxin/antitoxin system for preventing horizontal gene transfer.  
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This construct is composed of L-arabinose mediated inducible promoter pBAD and toxin CcdB. CcdB/CcdA is a type II toxin-antitoxin system originating from E. coli. In our project, we used this construct to test the toxicity of toxin CcdB, which provided us with more insights in regulating the balance of the toxin-antitoxin system for preventing horizontal gene transfer.  
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<!-- Add more about the biology of this part here
 
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<h2><span class="mw-headline" id="2021 Team Leiden">2021 Team Leiden</span><span class="mw-editsection"></span></h2>
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https://2021.igem.org/Team:Leiden
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CcdB has also been used as a part of a kill switch (Fig. 1) to test the toxicity of the toxin based on arabinose concentration.
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[[Image:T-Leiden-parts-Figuer4.png|500px]]
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'''''Figure 1.''''' ''Plate reader assay of E. coli TOP 10 cellular growth over time with a gradient of arabinose concentrations. ''
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Cell growth is strongly inhibited following the induction of CcdB. The results show that CcdB is not necessarily enough to kill all cells immediately when grown in liquid LB medium. Only after the exponential growth phase, the cells start to die. High concentrations of arabinose are needed for this killing effect, but that might change in different medium conditions.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3962342 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3962342 SequenceAndFeatures</partinfo>

Revision as of 07:50, 16 October 2021


arabinose-mediated inducible expression of toxin CcdB

This construct is composed of L-arabinose mediated inducible promoter pBAD and toxin CcdB. CcdB/CcdA is a type II toxin-antitoxin system originating from E. coli. In our project, we used this construct to test the toxicity of toxin CcdB, which provided us with more insights in regulating the balance of the toxin-antitoxin system for preventing horizontal gene transfer.


2021 Team Leiden

https://2021.igem.org/Team:Leiden 

CcdB has also been used as a part of a kill switch (Fig. 1) to test the toxicity of the toxin based on arabinose concentration.

T-Leiden-parts-Figuer4.png

Figure 1. Plate reader assay of E. coli TOP 10 cellular growth over time with a gradient of arabinose concentrations.

Cell growth is strongly inhibited following the induction of CcdB. The results show that CcdB is not necessarily enough to kill all cells immediately when grown in liquid LB medium. Only after the exponential growth phase, the cells start to die. High concentrations of arabinose are needed for this killing effect, but that might change in different medium conditions.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 1902
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1352
    Illegal SapI site found at 961