Part:BBa_K1632001
fim switch[default OFF](Tokyo_Tech/J23119)
The fim 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 fim switch by adding the Fim recombinase.
We designed this fim switch which has a J23119 promoter (BBa_J23119). Also between the promoter and the inverting site, there are two restriction enzyme cut sites are added in each side of the promoter (Fig. 1.). So the promoter can easily be interchanged. For example,BBa_K1632005, we replaced J23119 promoter with Lac promoter(BBa_R0010) (Fig. 2.). Except for insertion of restriction enzyme sites, basically, the design of fim switch(Tokyo_Tech) is similar with fim switch(wild-type).
From our results of our assay, the inversion of fim switch(Tokyo_Tech/J23119) by FimB(wild-type) (BBa_K1632010)/FimE(wild-type) (BBa_K1632011) was not confirmed correctly. The FimB protein(wild-type) inverts fim switch from [ON] state to [OFF] state and from [OFF] state to [ON] state correctly(Fig. 3.). However the FimE protein(wild-type) didn't invert fim switch predominantly from [ON] state to [OFF] state. In the assay, the FimE protein(wild-type) inverts fim switch from [ON] state to [OFF] state and from [OFF] state to [ON] state(Fig. 4.). In other words, the FimE protein works as the FimB protein.
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 98
Illegal NheI site found at 121 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 92
Illegal BamHI site found at 133 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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