Difference between revisions of "Part:BBa K817033:Design"
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===Design Notes=== | ===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 | + | 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 promoter, 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 our part, FadR is constitutively expressed and could be repressed by adding oleic acid(a long-chain fatty acid), releasing the FadRs and thereby expressing mRFP. | 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 our part, FadR is constitutively expressed and could be repressed by adding oleic acid(a long-chain fatty acid), releasing the FadRs and thereby expressing mRFP. | ||
Revision as of 16:55, 26 September 2012
PfadBA-RBS-mRFP
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 656
Illegal AgeI site found at 768 - 1000COMPATIBLE 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 promoter, 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 our part, FadR is constitutively expressed and could be repressed by adding oleic acid(a long-chain fatty acid), releasing the FadRs and thereby expressing mRFP.
Source
PfadBA-RFP