Difference between revisions of "Part:BBa K2549062"

 
 
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<partinfo>BBa_K2549062 short</partinfo>
 
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This part is a surface-expressed EGFP which can be served as antigen to the SynNotch receptor.
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This part is a surface-expressed EGFP which can be served as antigen to the SynNotch receptor. CMV promotor is a constitutive promotor. CD8&alpha; signal peptide is used to guide synthesized fusion protein to pass the translocon<ref>https://en.wikipedia.org/wiki/Translocon</ref> into the endoplasmic reticulum<ref>https://en.wikipedia.org/wiki/Endoplasmic_reticulum</ref>, and the fusion protein will be later sugar modified in Golgi<ref>https://en.wikipedia.org/wiki/Golgi_apparatus</ref>, presented on the plasma membrane and located to the outside of the cell. Transmembrane region of PDGFR&beta; embeds surCD19 on the membrane. This part is copied from our viral vector which can be expressed in mammalian cells. It is served as one of our antigen to SynNotch receptors.
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===Usage and Biology===
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<partinfo>BBa_K2549062 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2549062 SequenceAndFeatures</partinfo>
  
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===Our characterization===
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We used fluorescence confocal microscopy to capture the EGFP fluorescence which be successfully expressed on the surface of 293T cell. For more details, please refer to [http://2017.igem.org/Team:Fudan/Basic_Part 2017 Fudan team basic part].
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[[File:surEGFP.jpg|none|300px|thumb|'''surEGFP.''' It was captured by confocal and showed that it is expressed on the plasma membrane of 293T cell. We present this data in 2017 iGEM.  And, this plasmid has been requested by others since then. ]]
  
 
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<partinfo>BBa_K2549062 parameters</partinfo>
 
<partinfo>BBa_K2549062 parameters</partinfo>
 
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===References===

Latest revision as of 00:39, 18 October 2018


CMV-CD8α-EGFP-PDGFRβ

This part is a surface-expressed EGFP which can be served as antigen to the SynNotch receptor. CMV promotor is a constitutive promotor. CD8α signal peptide is used to guide synthesized fusion protein to pass the translocon[1] into the endoplasmic reticulum[2], and the fusion protein will be later sugar modified in Golgi[3], presented on the plasma membrane and located to the outside of the cell. Transmembrane region of PDGFRβ embeds surCD19 on the membrane. This part is copied from our viral vector which can be expressed in mammalian cells. It is served as one of our antigen to SynNotch receptors.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 531
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 540
  • 1000
    COMPATIBLE WITH RFC[1000]


Our characterization

We used fluorescence confocal microscopy to capture the EGFP fluorescence which be successfully expressed on the surface of 293T cell. For more details, please refer to [http://2017.igem.org/Team:Fudan/Basic_Part 2017 Fudan team basic part].

surEGFP. It was captured by confocal and showed that it is expressed on the plasma membrane of 293T cell. We present this data in 2017 iGEM. And, this plasmid has been requested by others since then.


References

  1. ↑ https://en.wikipedia.org/wiki/Translocon
  2. ↑ https://en.wikipedia.org/wiki/Endoplasmic_reticulum
  3. ↑ https://en.wikipedia.org/wiki/Golgi_apparatus