Difference between revisions of "Part:BBa K817033:Design"

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PfadBA-RFP contains two parts (1)fatty acid-dependent promoter(pfad) and ribosomal binding site(RBS) (2)mRFP, a red fluorescent protein. The promoter pfad acts as a switch of downstream protein expression, and is regulated by the fatty acid-dependent repressor, FadR. mRFP plays a role as a reporter that indicates the expression or efficiency of the promoter.  
 
PfadBA-RFP contains two parts (1)fatty acid-dependent promoter(pfad) and ribosomal binding site(RBS) (2)mRFP, a red fluorescent protein. The promoter pfad acts as a switch of downstream protein expression, and is regulated by the fatty acid-dependent repressor, FadR. mRFP plays a role as a reporter that indicates the expression or efficiency of the promoter.  
  
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In the absence of fatty acids, FadR binds at a site downstream of the pfad promoter and represses transcription. When long chain fatty acids become available, they bind to FadR, and elicit a conformational change that releases the protein from DNA, thus removing the repression.  
+
In the absence of fatty acids, FadR binds at a site downstream of the pfad promoter and represses transcription. When long chain fatty acids become available, they bind to FadR, and elicit a conformational change that releases the protein from DNA, thus removing the repression[1].  
  
 
In our design, FadR is constitutively expressed and could be repressed by adding oleic acid(a long-chain fatty acid), releasing the FadR and thereby expressing mRFP.
 
In our design, FadR is constitutively expressed and could be repressed by adding oleic acid(a long-chain fatty acid), releasing the FadR and thereby expressing mRFP.

Revision as of 17:19, 26 September 2012

PfadBA-RBS-mRFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 656
    Illegal AgeI site found at 768
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

PfadBA-RFP contains two parts (1)fatty acid-dependent promoter(pfad) and ribosomal binding site(RBS) (2)mRFP, a red fluorescent protein. The promoter pfad acts as a switch of downstream protein expression, and is regulated by the fatty acid-dependent repressor, FadR. mRFP plays a role as a reporter that indicates the expression or efficiency of the promoter.

In the absence of fatty acids, FadR binds at a site downstream of the pfad promoter and represses transcription. When long chain fatty acids become available, they bind to FadR, and elicit a conformational change that releases the protein from DNA, thus removing the repression[1].

In our design, FadR is constitutively expressed and could be repressed by adding oleic acid(a long-chain fatty acid), releasing the FadR and thereby expressing mRFP.

Source

pfad:72bps, from

RBS:12bps, from BBa_K093005

mRFP:681bps, from BBa_K093005

References