Difference between revisions of "Part:BBa K1632013"

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<partinfo>BBa_K1632013 short</partinfo>
 
<partinfo>BBa_K1632013 short</partinfo>
  
FimE is Fim recombinase.<br>
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[[Image:Tokyo_Tech_arabinosefimEsummary.png |thumb|center|900px|<b>Fig. 1. </b>New plasmids we constructed to confirm the function of BBa_K1632013 plasmid for Decision making coli.]]<br>
FimE inverts the <i>fim</i> switch (example <partinfo>BBa_K1632000</partinfo>, <partinfo>BBa_K1632004</partinfo> etc...) from the ON state to OFF state.<br>
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The expression of this Fim recombinase is controlled by arabinose in BBa_K1632013.
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For more information, see [http://2015.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2015 wiki].
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<span style="margin-left: 10px;">The <i>fim</i> switch is inverted by FimE.The FimE protein inverts the ''fim'' switch predominantly in the ON-to-OFF direction.<br>
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<span style="margin-left: 10px;">In order to assay the function of our FimE, we added a GFP coding sequence on the downstream of the <i>fim</i> switch.The <i>fim</i> switch[default ON](wild-type)_gfp (BBa_K1632007) emitts fluorescence when expressed, while the <i>fim</i> switch [default OFF](wild-type)_gfp(BBa_K1632008) does not emit florescence when expressed. We also added PBAD/''araC'' on the upstream of fimE.PBAD/''araC''_fimE (BBa_K1632013) can induce the expression of FimE in the presence of arabinose. We co-transformed a <i>fim</i> switch-gfp and a PBAD/''araC''_fim recombinase in the E. coli DH5alpha strain. We measured the fluorescence intensity of the cells induced by different concentraions of arabinose.<br>
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[[Image:Tokyo_Tech_FimE_assay_Results.png |thumb|center|700px|<b>Fig. 5. </b>Histogram of the samples measured by flow cytometer]]<br>
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<span style="margin-left: 10px;">From the experimental results,our fimE inverted the <i>fim</i> switch[default ON](wild-type) from ON-to-OFF but did not invert the <i>fim</i> switch[default OFF](wild-type) from the OFF state, depending on the concentration of arabinose.<br><br><br>
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For more information, see [http://2015.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2015 wiki]. <br><br><br>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 07:32, 18 September 2015

PBAD/araC_rbs_fimE(wild-type)

Fig. 1. New plasmids we constructed to confirm the function of BBa_K1632013 plasmid for Decision making coli.

The fim switch is inverted by FimE.The FimE protein inverts the fim switch predominantly in the ON-to-OFF direction.


In order to assay the function of our FimE, we added a GFP coding sequence on the downstream of the fim switch.The fim switch[default ON](wild-type)_gfp (BBa_K1632007) emitts fluorescence when expressed, while the fim switch [default OFF](wild-type)_gfp(BBa_K1632008) does not emit florescence when expressed. We also added PBAD/araC on the upstream of fimE.PBAD/araC_fimE (BBa_K1632013) can induce the expression of FimE in the presence of arabinose. We co-transformed a fim switch-gfp and a PBAD/araC_fim recombinase in the E. coli DH5alpha strain. We measured the fluorescence intensity of the cells induced by different concentraions of arabinose.

Fig. 5. Histogram of the samples measured by flow cytometer

From the experimental results,our fimE inverted the fim switch[default ON](wild-type) from ON-to-OFF but did not invert the fim switch[default OFF](wild-type) from the OFF state, depending on the concentration of 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
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1260
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961