Difference between revisions of "Part:BBa K1632005"
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+ | [[Image:Tokyo Tech fim switch (wild-type) design.png|thumb|center|400px|Fig. 1. Overview of fim inversion system]]<br> | ||
<span style="margin-left: 10px;">''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)(<partinfo>BBa_K1632004</partinfo>) transcripts to the right.<br> | <span style="margin-left: 10px;">''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)(<partinfo>BBa_K1632004</partinfo>) transcripts to the right.<br> |
Revision as of 01:59, 18 September 2015
fim switch[default OFF](wild-type)
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 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 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]