Difference between revisions of "Part:BBa K1632005"

Revision as of 21:55, 18 September 2015

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.

Sequence and Features