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

(→‎Source)
(→‎Design Notes)
 
(3 intermediate revisions by one other user not shown)
Line 8: Line 8:
 
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.  
  
−
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.
 +
 +
For functional assay result, please refer to our [http://2012.igem.org/Team:NTU-Taida/Result/pFadBA wiki page].
  
 
===Source===
 
===Source===
Line 16: Line 18:
  
  
−
pfad:72bps, from
+
pfad:72bps, from de novo synthesis
  
 
RBS:12bps, from BBa_K093005
 
RBS:12bps, from BBa_K093005
Line 23: Line 25:
  
 
===References===
 
===References===
 +
 +
[1] The FadR.DNA complex. Transcriptional control of fatty acid metabolism in Escherichia coli. ''J Biol Chem. 2001 May 18;276(20):17373-9''

Latest revision as of 01:43, 27 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.

For functional assay result, please refer to our [http://2012.igem.org/Team:NTU-Taida/Result/pFadBA wiki page].

Source

pfad:72bps, from de novo synthesis

RBS:12bps, from BBa_K093005

mRFP:681bps, from BBa_K093005

References

[1] The FadR.DNA complex. Transcriptional control of fatty acid metabolism in Escherichia coli. J Biol Chem. 2001 May 18;276(20):17373-9