Regulatory

Part:BBa_K1632005

Designed by: Riku Shinohara   Group: iGEM15_Tokyo_Tech   (2015-08-30)
Revision as of 00:31, 19 September 2015 by JunKawamura (Talk | contribs)

fim switch[default OFF](wild-type)

Fig. 1. Overview of fim inversion system

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 left. From the direction of transcription, the state is defined as [OFF]. On the other hand, fim switch[default ON](wild-type)(BBa_K1632004) transcripts to the right.

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

We constructed fim switch[default OFF](wild-type)_rbs_gfp(BBa_K1632008) 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[default OFF](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 OFF](wild-type)_rbs_gfp and PBAD/araC_fimE(wild-type)(BBa_K1632013) in the E.coli DH5alpha strain.We measured the fluorescence intensity of the cells induced by arabinose.

More information

For more information, see http://2015.igem.org/Team:Tokyo_Tech/Project Our work in Tokyo_Tech 2015 wiki, http://2015.igem.org/Team:Tokyo_Tech/Experiment/ssrA_tag_degradation_assay About ssrA-tag, http://2015.igem.org/Team:Tokyo_Tech/Experiment/Overview_of_fim_inversion_system About ''fim'' inversion system

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]


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