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

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

@@ Line 11: / Line 11: @@
 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===