Composite

Part:BBa_K1632007

Designed by: Riku Shinohara   Group: iGEM15_Tokyo_Tech   (2015-08-30)
Revision as of 14:42, 18 September 2015 by Y.Yuta (Talk | contribs)

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

File:Tokyo Tech fim switch(wild-type) design.png
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 in [ON] state to [OFF] state and [OFF] to [ON] state with approximately equal efficiencies. On the other hand, the FimE protein inverts fim switch predominantly [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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1052


[edit]
Categories
//awards/part_collection/2015
//function/recombination/fim
Parameters
None