Difference between revisions of "Part:BBa K1632007"

Revision as of 15:08, 18 September 2015

fim switch[default ON](wild-type)_rbs_gfp

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 constitutive promoter. The promoter in the fim switch transcripts to the right. From the direction of transcription, the state is defined as [ON] state. 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 from [ON] state to [OFF] state and from [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 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