Difference between revisions of "Part:BBa K2549062"
Line 4: Line 4:
  
 
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.
 
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.
−
 
−
 
  
 
<!-- -->
 
<!-- -->
Line 15: Line 13:
 
We used fluorescence microscopy to capture the EGFP which be successfully expressed on the surface of 293T cell.
 
We used fluorescence microscopy to capture the EGFP which be successfully expressed on the surface of 293T cell.
  
−
[[File:surEGFP.jpg|none|540px|thumb|'''Fig.1 EGFP is detected to be expressed on the plasma membrane of 293T cell.''']]
+
[[File:surEGFP.jpg|none|360px|thumb|'''Fig.1 EGFP is detected to be expressed on the plasma membrane of 293T cell.''']]
 +
 
 +
For more details, please refer to our wiki <b>[http://2017.igem.org/Team:Fudan/Basic_Part 2017 Fudan team basic part]</b>
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 14:06, 16 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]

Characterization

We used fluorescence microscopy to capture the EGFP which be successfully expressed on the surface of 293T cell.

Fig.1 EGFP is detected to be expressed on the plasma membrane of 293T cell.

For more details, please refer to our wiki [http://2017.igem.org/Team:Fudan/Basic_Part 2017 Fudan team basic part]


References

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