Difference between revisions of "Part:BBa K817001:Design"
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The first experiment we conducted showed that the real situation might not be what we expected, so we decided to clone out the FadR gene and construct it into our circuit. We prepared our FadR gene by doing the PCR of the genomic DNA of E. coli. K-12 DH5α strain. Later, we incorporated the FadR gene we cloned into BBa_J04500, with plac(R0010) and RBS(B0034) to test the function, forming BBa_K817007 part. We transformed both our pfad-mRFP part(BBa_K817033) and BBa_K817007 part into E. coli. K-12 DH5α to do the test. Functional results can be found in the result website of 2012 iGEM_Taida(http://2012.igem.org/Team:NTU-Taida/Result)
 
The first experiment we conducted showed that the real situation might not be what we expected, so we decided to clone out the FadR gene and construct it into our circuit. We prepared our FadR gene by doing the PCR of the genomic DNA of E. coli. K-12 DH5α strain. Later, we incorporated the FadR gene we cloned into BBa_J04500, with plac(R0010) and RBS(B0034) to test the function, forming BBa_K817007 part. We transformed both our pfad-mRFP part(BBa_K817033) and BBa_K817007 part into E. coli. K-12 DH5α to do the test. Functional results can be found in the result website of 2012 iGEM_Taida(http://2012.igem.org/Team:NTU-Taida/Result)
  
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The FadR gene had already been sequenced and compared. The result shows 100%-fit with the sequence in the NCBI FadR gene sequence database of E. coli. K-12 genome.
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The FadR gene had already been sequenced and compared. The result shows 100%-fit with the sequence in the NCBI FadR gene sequence database of ''E. coli''. K-12 genome.
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+
−
 
+
  
 
===Source===
 
===Source===
  
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from E.coli K12 DH5α genome. Cloned by PCR, 2012 iGEM_Taida, National Taiwan University, Taipei, Taiwan. Confirmed by sequence.
+
from ''Escherichia coli.'' K12 DH5α genome. Cloned by PCR, 2012 iGEM_Taida, National Taiwan University, Taipei, Taiwan. Confirmed by sequence.
  
 
===References===
 
===References===
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NCBI database for reference FadR protein sequence.
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DNA-binding transcriptional dual regulator of fatty acid metabolism [Escherichia coli str. K-12 substr. MG1655]

Latest revision as of 02:41, 27 September 2012

FadR


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

One of our team's goal is to design a fatty-acid-detecting system, using the natural fatty-acid sensing genes, including fatty acid sensing promotor, pfadBA and the natural repressor, FadR.

At first, from the papers we reviewed, we assumed that FadR gene within the Escherichia coli(E. coli) bacteria will be normally expressed to a certain content that pfad promotor will be repressed. The first experiment we conducted showed that the real situation might not be what we expected, so we decided to clone out the FadR gene and construct it into our circuit. We prepared our FadR gene by doing the PCR of the genomic DNA of E. coli. K-12 DH5α strain. Later, we incorporated the FadR gene we cloned into BBa_J04500, with plac(R0010) and RBS(B0034) to test the function, forming BBa_K817007 part. We transformed both our pfad-mRFP part(BBa_K817033) and BBa_K817007 part into E. coli. K-12 DH5α to do the test. Functional results can be found in the result website of 2012 iGEM_Taida(http://2012.igem.org/Team:NTU-Taida/Result)

The FadR gene had already been sequenced and compared. The result shows 100%-fit with the sequence in the NCBI FadR gene sequence database of E. coli. K-12 genome.

Source

from Escherichia coli. K12 DH5α genome. Cloned by PCR, 2012 iGEM_Taida, National Taiwan University, Taipei, Taiwan. Confirmed by sequence.

References

NCBI database for reference FadR protein sequence. DNA-binding transcriptional dual regulator of fatty acid metabolism [Escherichia coli str. K-12 substr. MG1655]