Difference between revisions of "Part:BBa K1632001"

Revision as of 13:07, 16 September 2015

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 sites in each front (SalIand BamHI) and back (BglII and MluI)(Fig. 1. Tokyo Tech fim switch (Tokyo_Tech/J23119) design). So the promoter can easily be interchanged. Except for insertion of restriction enzyme sites, basically, the design of TokyoTech fim switch is similar with Wild Type fim switch.

Fig. 1. Tokyo Tech fim switch (Tokyo_Tech/J23119) design (Up:on state Down:off state)

We constructed fim switch [default OFF](Tokyo_Tech/J23119)_rbs_gfp(BBa_K1632003) to characterize the function of this part, by inserting this promoter upstream of a GFP coding sequence.
From our results of our assay, the inversion of fim switch (Tokyo_Tech/J23119) by FimB/FimE was not confirmed correctly. The FimB protein inverts fim switch in the ON-to-OFF and the OFF-to-ON direction correctly. However the FimE protein didn’t inverts fim switch predominantly in the ON-to-OFF direction. In the assay, the FimE protein inverts fim switch in the ON-to-OFF and the OFF-to-ON direction. In other words, the FimE protein works as the FimB protein.

Sequence and Features