Difference between revisions of "Part:BBa K1632007"

Revision as of 13:25, 16 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 constitutively promoter. The promoter in the fim switch transcripts to the right. From the direction of transcription, the state is defined as ON. 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 in the ON-to-OFF and the OFF-to-ON direction with approximately equal efficiencies. On the other hand, the FimE protein inverts fim switch predominantly in the ON-to-OFF direction.

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.

Sequence and Features