Difference between revisions of "Part:BBa K1632004"
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<partinfo>BBa_K1632004 short</partinfo>
 
<partinfo>BBa_K1632004 short</partinfo>
  
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The <i>fim</i> switch is the promoter containing repeated DNA sequence which is inverted by the Fim recombinase. Therefore, we can control the expression of the gene downstream of the <i>fim</i> switch by adding the Fim recombinase.<br>
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''Fim'' switch (wild-type) is derived from wild-type sequence. ''Fim'' switch (wild type) have sigma 70 promoter which is constitutively promoter. The promoter in the ''fim'' switch transcripts to the right. From the direction of transcription, the state is defined as ON.<br>
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This gene sequence is the same as that of a wild type E.coli.
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''Fim'' switch is inverted by two recombinase, FimB and FimE.The FimB protein inverts ''fim'' switch in the ON-to-OFF and the OFF-to-ON direction with approximately equal efficiencies. On the other hand, the FimE protein inverts ''fim'' switch predominantly in the ON-to-OFF direction.<br>
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We constructed ''fim'' switch[ON](wild-type)-rbs-gfp(<partinfo>BBa_K1632007</partinfo>) to characterize the function of this part, by inserting this promoter upstream of a GFP coding sequence. <br>
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In the fimB dependent ''fim'' switch state assay, we transformed ''fim'' switch[ON](wild-type)-rbs-gfp and pBAD/araC-fimB(<partinfo>BBa_K1632012</partinfo>)in the E.coli DH5alfa strain. We measured the fluorescence intensity of the cells induced by arabinose. <br>
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Similary,in the fimE dependent ''fim'' switch state assay, we transformed ''fim'' switch[ON](wild-type)-rbs-gfp and pBAD/araC-fimE(<partinfo>BBa_K1632013</partinfo>) in the E.coli DH5alfa strain.We measured the fluorescence intensity of the cells induced by arabinose. <br><br>
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Revision as of 19:05, 13 September 2015

fim switch[default ON](wild-type)

Fim switch (wild-type) is derived from wild-type sequence. Fim switch (wild type) have sigma 70 promoter which is constitutively promoter. The promoter in the fim switch transcripts to the right. From the direction of transcription, the state is defined as ON.


Fim switch is inverted by two recombinase, FimB and FimE.The FimB protein inverts fim switch in the ON-to-OFF and the OFF-to-ON direction with approximately equal efficiencies. On the other hand, the FimE protein inverts fim switch predominantly in the ON-to-OFF direction.


We constructed fim switch[ON](wild-type)-rbs-gfp(BBa_K1632007) to characterize the function of this part, by inserting this promoter upstream of a GFP coding sequence.

In the fimB dependent fim switch state assay, we transformed fim switch[ON](wild-type)-rbs-gfp and pBAD/araC-fimB(BBa_K1632012)in the E.coli DH5alfa strain. We measured the fluorescence intensity of the cells induced by arabinose.

Similary,in the fimE dependent fim switch state assay, we transformed fim switch[ON](wild-type)-rbs-gfp and pBAD/araC-fimE(BBa_K1632013) in the E.coli DH5alfa strain.We measured the fluorescence intensity of the cells induced by arabinose.


Sequence and Features


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]