Difference between revisions of "Part:BBa K3064014:Design"
 
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Choosing hsaGlucagon as the antibody in our project because of the specificity between antibody and antigen. Linking the hsaGlucagon  and the hsalgG-Fc makes a promising recombinant antibody that is both recognizable by trim21.
 
Choosing hsaGlucagon as the antibody in our project because of the specificity between antibody and antigen. Linking the hsaGlucagon  and the hsalgG-Fc makes a promising recombinant antibody that is both recognizable by trim21.
 
P2A in our design aims to achieve the automatic cleavage of two peptides, offering a convenient way to express two proteins only with one promoter. <Br/>
 
P2A in our design aims to achieve the automatic cleavage of two peptides, offering a convenient way to express two proteins only with one promoter. <Br/>
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4XGS is the improvement we made this year, based on a 3XGS linker(BBa_K1486004). Besides the increased length, we made changes on the sequence of the middle GGGGS, so that the 3XGS could be more flexible and more specific in PCR and other aspects.
 
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===Source===
 
===Source===

Latest revision as of 09:01, 29 September 2019


hsaGlucagon->GGGGS->hsaIgG-Fc->P2A->HA->hsaTrim21


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1830
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2499
    Illegal BamHI site found at 3037
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1787
    Illegal AgeI site found at 632
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1142
    Illegal BsaI site found at 2877
    Illegal SapI.rc site found at 1490


Design Notes

Choosing hsaGlucagon as the antibody in our project because of the specificity between antibody and antigen. Linking the hsaGlucagon and the hsalgG-Fc makes a promising recombinant antibody that is both recognizable by trim21. P2A in our design aims to achieve the automatic cleavage of two peptides, offering a convenient way to express two proteins only with one promoter.

Source

1-The sequence of trim21 comes from the article: James L C, Keeble A H, Khan Z, et al. Structural basis for PRYSPRY-mediated tripartite motif (TRIM) protein function[J]. Proceedings of the National Academy of Sciences of the United States of America, 2007, 104(15):6200-6205.
2-The sequence of P2A comes from the article:Jin H K, Lee S R, Li L H, et al. High Cleavage Efficiency of a 2A Peptide Derived from Porcine, Teschovirus-1 in Human Cell Lines, Zebrafish and Mice[J]. Plos One, 2011, 6(4):e18556.

References

[1]Clift D, Mcewan W A, Labzin L I, et al. A Method for the Acute and Rapid Degradation of Endogenous Proteins[J]. Cell, 2017, 171(7):1692-1706.e18.
[2]Mcewan W A, Falcon B, Vaysburd M, et al. Cytosolic Fc receptor TRIM21 inhibits seeded tau aggregation.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2017, 114(3):574.