Difference between revisions of "Part:BBa K1632008"
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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 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) (<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 constitutively 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 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.<br>
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<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 [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>
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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>
  
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<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>
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<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>
  
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<span style="margin-left: 10px;">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)(<partinfo>BBa_K1632013</partinfo>) in the E.coli DH5alfpha strain.We measured the fluorescence intensity of the cells induced by arabinose. <br><br>
+
<span style="margin-left: 10px;">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)(<partinfo>BBa_K1632013</partinfo>) in the E.coli DH5alfpha strain.We measured the fluorescence intensity of the cells induced by arabinose. <br><br>
  
 
For more information, see [http://2015.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2015 wiki].
 
For more information, see [http://2015.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2015 wiki].

Revision as of 22:25, 18 September 2015

fim switch[default OFF](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] 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 [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