Difference between revisions of "Part:BBa K1073028"

 
 
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<partinfo>BBa_K1073028 short</partinfo>
 
<partinfo>BBa_K1073028 short</partinfo>
  
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In complex with the inducer N-butyryl-HSL, the activator RhlR binds to the Rhl promoter and enables expression. This construct provides all the parts required to express the gene behind the rhl promoter. Only the inducer N-butyryl-HSL is missing, which usually is taken up from the media. By the regulation of the inducer's concentration in the media, the expression rate of the gene of interest (in this case ampicillin beta-lactamase) can be controlled.  
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Induction of the beta lactamase mediated ampicillin resistance by RhlR and specific autoinducer N-butyryl homoserine lactone.
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In complex with the autoinducer molecule N-butyryl-HSL, the transcription regulator RhlR binds to specific sequences in the promoter region and enables transcription of ''ampR''.
  
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===Usage and Biology===
 
===Usage and Biology===
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The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-butyryl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-buturyl-HSL to the culture broth.
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Expression of ''RhlR'' is regulated by the strong constitutive promoter <bbpart>BBa_J23100</bbpart> and RBS of medium strengh (<bbpart>BBa_B0032</bbpart>).
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Note that due to the low level leakiness of the promoter there is background activity of the beta lactamase even when N-butyryl-HSL is not present.
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Latest revision as of 09:28, 17 October 2013

Inducible ampicillin resistance cassette + RhlR expression cassette

Induction of the beta lactamase mediated ampicillin resistance by RhlR and specific autoinducer N-butyryl homoserine lactone.

In complex with the autoinducer molecule N-butyryl-HSL, the transcription regulator RhlR binds to specific sequences in the promoter region and enables transcription of ampR.

Usage and Biology

The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-butyryl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-buturyl-HSL to the culture broth. Expression of RhlR is regulated by the strong constitutive promoter BBa_J23100 and RBS of medium strengh (BBa_B0032).

Note that due to the low level leakiness of the promoter there is background activity of the beta lactamase even when N-butyryl-HSL is not present.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1096
    Illegal NheI site found at 1119
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1391
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1866