Difference between revisions of "Part:BBa K1073030"

 
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<partinfo>BBa_K1073030 short</partinfo>
 
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In complex with the inducer PAI-1 (N-(3-oxododecanoyl)-homoserine lactone), the activator LasR binds to the Las promoter and enables expression. This construct provides all the parts required to express the gene behind the Las promoter. Only the inducer N-(3-oxododecanoyl)-homoserine lactone 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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In complex with the autoinducer molecule N-3-oxododecanoyl homoserine lactone (N-3-oxododecanoyl-HSL) the transcription regulator LasR binds to specific sequences in the promoter region and enables transcription of ''ampR''. This construct provides all the parts required to express the gene downstream of the inducible promoter. Only LasI, the synthase producing the autoinducer molecule N-3-oxododecanoyl-HSL is missing in the construct. However, N-3-oxododecanoyl-HSL can be taken up from the media.  
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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-buturyl-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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Note that due to the leakiness of the promoter there is background activity of the beta lactamase even when N-3-oxododecanoyl-HSL is not present.
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Revision as of 17:41, 24 September 2013

Inducible ampicillin resistance cassette + LasR expression cassette

In complex with the autoinducer molecule N-3-oxododecanoyl homoserine lactone (N-3-oxododecanoyl-HSL) the transcription regulator LasR binds to specific sequences in the promoter region and enables transcription of ampR. This construct provides all the parts required to express the gene downstream of the inducible promoter. Only LasI, the synthase producing the autoinducer molecule N-3-oxododecanoyl-HSL is missing in the construct. However, N-3-oxododecanoyl-HSL can be taken up from the media.


Usage and Biology

The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-buturyl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-buturyl-HSL to the culture broth.

Note that due to the leakiness of the promoter there is background activity of the beta lactamase even when N-3-oxododecanoyl-HSL is not present.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1169
    Illegal NheI site found at 1192
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1546
    Illegal AgeI site found at 1743
  • 1000
    COMPATIBLE WITH RFC[1000]