Difference between revisions of "Part:BBa K533005"

 
 
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<partinfo>BBa_K533005 short</partinfo>
 
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GFP is presented outside E. Coli as a positive control for protein presentation
 
GFP is presented outside E. Coli as a positive control for protein presentation
  
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===Usage and Biology===
 
===Usage and Biology===
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This is a surface marker for E. coli, although it can serve other purposes.
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We verified its membrane location by protease digestion and Western blotting.
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{| align="center" style="border: none; background: none; border-spacing: 0;text-align:center;margin-left: 100px;"
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! style="border: 1px solid #39F;padding: 5px;" |  Color Channel
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! style="border: 1px solid #39F;padding: 5px;" |  Post-digest
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! style="border: 1px solid #39F;padding: 5px;" |  Pre-digest
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|-
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|  style="border: 1px solid #39F;padding: 5px;" | GFP
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| style="border: 1px solid #39F;padding: 5px;" |  [[Image:Thuexp_gn.png|140px]]
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| style="border: 1px solid #39F;padding: 5px;" |  [[Image:Thuexp_gp.png|140px]]
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|-
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| style="border: 1px solid #39F;padding: 5px;" |  DAPI
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| style="border: 1px solid #39F;padding: 5px;" |  [[Image:Thuexp_gn_d.png|140px]]
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| style="border: 1px solid #39F;padding: 5px;" |  [[Image:Thuexp_gp_d.png|140px]]
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|-
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| style="border: 1px solid #39F;padding: 5px;" |  Merge
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| style="border: 1px solid #39F;padding: 5px;" |  [[Image:Thuexp_gn_m.png|140px]]
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| style="border: 1px solid #39F;padding: 5px;" |  [[Image:Thuexp_gp_m.png|140px]]
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|}
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Disappearance of green fluorescence suggests that GFP is predominantly on the outside of E. coli.
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Fractionation and western blotting also suggests that the protein is predominantly on the membrane.
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[[Image:Thuwb.png]]
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Latest revision as of 15:44, 5 October 2011

OmpA-GFP

GFP is presented outside E. Coli as a positive control for protein presentation

Usage and Biology

This is a surface marker for E. coli, although it can serve other purposes.

We verified its membrane location by protease digestion and Western blotting.

Color Channel Post-digest Pre-digest
GFP Thuexp gn.png Thuexp gp.png
DAPI Thuexp gn d.png Thuexp gp d.png
Merge Thuexp gn m.png Thuexp gp m.png

Disappearance of green fluorescence suggests that GFP is predominantly on the outside of E. coli.

Fractionation and western blotting also suggests that the protein is predominantly on the membrane.

Thuwb.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1419
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]