Part:BBa_K1632007
fim switch[default ON](wild-type)_rbs_gfp
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. On the other hand, fim switch [default OFF](wild-type) (BBa_K1632005) transcripts to the left.
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 [default ON](wild-type)_rbs_gfp(BBa_K1632007) to characterize the function of this part, by inserting fim switch [default ON](wild-type)(BBa_K1632004) upstream of a GFP coding sequence.
In the fimB dependent fim switch state assay, we transformed fim switch[default ON](wild-type)_rbs_gfp and PBAD/araC_fimB(BBa_K1632012)in the E.coli DH5alpha 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[default ON](wild-type)_rbs_gfp and PBAD/araC_fimE(wild-type)(BBa_K1632013) in the E.coli DH5alfpha strain.We measured the fluorescence intensity of the cells induced by arabinose.
For more information, see [http://2015.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2015 wiki].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1052
//function/recombination/fim
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