Difference between revisions of "Part:BBa K3788019"

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https://2021.igem.org/wiki/images/9/98/T--Aix-Marseille--BBa_K3788019.jpeg
 
https://2021.igem.org/wiki/images/9/98/T--Aix-Marseille--BBa_K3788019.jpeg
 
<i><b> Figure :</b> schematic representation of the construction with the different coding sequences, restriction sites used to insert the GFP coding sequence, and the known terminators.</i>
 
<i><b> Figure :</b> schematic representation of the construction with the different coding sequences, restriction sites used to insert the GFP coding sequence, and the known terminators.</i>
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===Methods===
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<b>METTRE LE MEME PROTOCOLE QUE POUR LA PART 017</b>
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===Results===
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https://2021.igem.org/wiki/images/7/73/T--Aix-Marseille--Activity_of_the_LexA-dependent_promoter_fused_to_GFP.jpeg
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<i><b>Figure:</b>Measure of the GFP signal (as fluorescein equivalent) of bacterial liquid cultures expriming BBa_K3788019 across time with different quantities of MitomycinC added to the growing medium.</i>
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<p><i>The experiment was done in triplicate. Standard deviation is small and not visible on the graph.</i></p>
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<p>We can see that GFP is expressed in response to the presence of MitomycinC in the cultures, and moreover we observe that the highest the quantity of mitomycin is, the highest the GFP signal is. It gives us the information that the BBa_K3788017 works with a dose-dependant manner.</p>
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Revision as of 22:53, 20 October 2021


Expression of GFP and of the timer device under the control of the LexA-controlled promoter



Induced by the SOS response (LexA dependent) upon DNA damages (UV, mitomycin…). This promoter allow the expression of the operon caa and cal. caa gene is coding for Colicin A (ColA) a toxin toxin produced by E. coli strains, and active against other non-immune E.coli strains. In this contruction GFP permit to understand the mechanism, and ColA isn't able to have his toxin activity. To protect themselfs, E. coli have an imumnity protein coded by cai gene (-). cal gene is codding for a lysis protein, when this protein is produce ColA can be delivered in the extracellular medium after the E. coli cells lysis. The regulation is complex and involves many regulatory elements: terminator (+) and (-), promoters, antisense coding sequence, etc.

T--Aix-Marseille--BBa_K3788019.jpeg Figure : schematic representation of the construction with the different coding sequences, restriction sites used to insert the GFP coding sequence, and the known terminators.

Methods

METTRE LE MEME PROTOCOLE QUE POUR LA PART 017

Results

T--Aix-Marseille--Activity_of_the_LexA-dependent_promoter_fused_to_GFP.jpeg

Figure:Measure of the GFP signal (as fluorescein equivalent) of bacterial liquid cultures expriming BBa_K3788019 across time with different quantities of MitomycinC added to the growing medium.

The experiment was done in triplicate. Standard deviation is small and not visible on the graph.


We can see that GFP is expressed in response to the presence of MitomycinC in the cultures, and moreover we observe that the highest the quantity of mitomycin is, the highest the GFP signal is. It gives us the information that the BBa_K3788017 works with a dose-dependant manner.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1528
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1848
    Illegal BamHI site found at 1521
    Illegal XhoI site found at 1397
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1597
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 867
    Illegal SapI.rc site found at 1351