Difference between revisions of "Part:BBa K1632007"
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[[Image:Tokyo Tech fim switch (wild-type) design.png|thumb|center|400px|Fig. 1. Overview of fim inversion system]]<br>
 
[[Image:Tokyo Tech fim switch (wild-type) design.png|thumb|center|400px|Fig. 1. Overview of fim inversion system]]<br>
  
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<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 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) (<partinfo>BBa_K1632005</partinfo>) transcripts to the left.<br>
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<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 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) (<partinfo>BBa_K1632005</partinfo>) transcripts to the left.<br>
  
 
<span style="margin-left: 10px;">''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.<br>
 
<span style="margin-left: 10px;">''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.<br>
  
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<span style="margin-left: 10px;">We constructed ''fim'' switch [default ON](wild-type)_rbs_gfp(<partinfo>BBa_K1632007</partinfo>) to characterize the function of this part, by inserting ''fim'' switch [default ON](wild-type)(<partinfo>BBa_K1632004</partinfo>) upstream of a GFP coding sequence. <br>
+
<span style="margin-left: 10px;">We constructed ''fim'' switch[default ON](wild-type)_rbs_gfp(<partinfo>BBa_K1632007</partinfo>) to characterize the function of this part, by inserting ''fim'' switch[default ON](wild-type)(<partinfo>BBa_K1632004</partinfo>) upstream of a GFP coding sequence. <br>
  
 
<span style="margin-left: 10px;">In the fimB dependent ''fim'' switch state assay, we transformed ''fim'' switch[default ON](wild-type)_rbs_gfp and PBAD/araC_fimB(<partinfo>BBa_K1632012</partinfo>) in the E.coli DH5alpha strain. We measured the fluorescence intensity of the cells induced by arabinose. <br>
 
<span style="margin-left: 10px;">In the fimB dependent ''fim'' switch state assay, we transformed ''fim'' switch[default ON](wild-type)_rbs_gfp and PBAD/araC_fimB(<partinfo>BBa_K1632012</partinfo>) in the E.coli DH5alpha strain. We measured the fluorescence intensity of the cells induced by arabinose. <br>

Revision as of 15:07, 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


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