Difference between revisions of "Part:BBa K523013"
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__NOTOC__
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<partinfo>BBa_K523013 short</partinfo>
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[[Image:Edinburgh-INP-YFP-cells.jpg|thumb|right]]
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Fusion of Ice Nucleation Protein (INP) and Enhanced Yellow Fluorescent Protein (EYFP) under the control of the Lac promoter.
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Constructed using the BioSandwich protocol of Edinburgh 2011.
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===Usage and Biology===
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Cells expressing this construct are shown on the right. The cells fluoresce yellow under blue light. The INP should carry the EYFP to the outer membrane of ''E. coli''. Our imaging technology was not good enough to confirm this, so we attempted to prove it by other means...
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<br clear="all" />
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====Fractionation experiment====
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We (Edinburgh 2011) centrifuged cells expressing BBa_K523013 so that the membrane fraction became localised to the bottom of the tube, and compared the fluorescence pattern with a control where EYFP was not fused to INP.
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<center>
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{| style="margin-top: 1em; margin-bottom: 1em;"
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|-
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|[[Image:523013-centrifuged.jpg|200px]]
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|&nbsp; &nbsp;
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|[[Image:523013-centrifuged-auto-white.jpg|200px]]
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|-
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|width="200px" valign="top" | Control on left. '''INP-EYFP on right.'''
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|&nbsp; &nbsp;
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|width="200px" valign="top" | The same image passed through GIMP's auto white balance filter.
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|}
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</center>
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These results appear to indicate that the EYFP was successfully transported to the outer membrane; this region is the main source of fluorescence in the INP-EYFP tube. By contrast, the control tube has much more EYFP present in the cytoplasm.
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<span class='h3bb'>'''Sequence and Features'''</span>
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<partinfo>BBa_K523013 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K523013 parameters</partinfo>
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<!-- -->

Revision as of 16:17, 10 May 2013

Plac + INP-EYFP

Edinburgh-INP-YFP-cells.jpg

Fusion of Ice Nucleation Protein (INP) and Enhanced Yellow Fluorescent Protein (EYFP) under the control of the Lac promoter.

Constructed using the BioSandwich protocol of Edinburgh 2011.

Usage and Biology

Cells expressing this construct are shown on the right. The cells fluoresce yellow under blue light. The INP should carry the EYFP to the outer membrane of E. coli. Our imaging technology was not good enough to confirm this, so we attempted to prove it by other means...


Fractionation experiment

We (Edinburgh 2011) centrifuged cells expressing BBa_K523013 so that the membrane fraction became localised to the bottom of the tube, and compared the fluorescence pattern with a control where EYFP was not fused to INP.

523013-centrifuged.jpg     523013-centrifuged-auto-white.jpg
Control on left. INP-EYFP on right.     The same image passed through GIMP's auto white balance filter.

These results appear to indicate that the EYFP was successfully transported to the outer membrane; this region is the main source of fluorescence in the INP-EYFP tube. By contrast, the control tube has much more EYFP present in the cytoplasm.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1036
  • 1000
    COMPATIBLE WITH RFC[1000]