Difference between revisions of "Part:BBa K1073030"

(→‎Induction of expression by RhlR and N-3-oxododecanoyl-HSL)
 
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Induction of the beta lactamase mediated ampicillin resistance by LasR and specific autoinducer N-3-oxododecanoyl homoserine lactone.
 
Induction of the beta lactamase mediated ampicillin resistance by LasR and specific autoinducer N-3-oxododecanoyl homoserine lactone.
 
   
 
   
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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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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''.
  
  
 
===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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The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-3-oxododecanoyl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-3-oxododecanoyl-HSL to the culture broth. Expression of ''lasR'' is regulated by the strong constitutive promoter <bbpart>BBa_J23100</bbpart>  and RBS of medium strengh (<bbpart>BBa_B0032</bbpart>). 
  
 
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.  
 
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.  

Latest revision as of 09:32, 17 October 2013

Inducible ampicillin resistance cassette + LasR expression cassette


Induction of the beta lactamase mediated ampicillin resistance by LasR and specific autoinducer N-3-oxododecanoyl homoserine lactone.

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.


Usage and Biology

The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-3-oxododecanoyl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-3-oxododecanoyl-HSL to the culture broth. Expression of lasR is regulated by the strong constitutive promoter BBa_J23100 and RBS of medium strengh (BBa_B0032).

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]