Difference between revisions of "Part:BBa K2653006:Design"

 
(→‎References)
 
(2 intermediate revisions by the same user not shown)
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
ddddd
+
Choosing nanobody as the ideal antibody in our project aims to avoid the disadvantage of traditional antibodies, among which the most difficult is the large sequence amount of these antibodies that may influence the transfection or the expression of the part. Linking the recombinate scFv of erbB3 and the hIgG1-Fc makes a promising recombinant antibody that is both recognizable by trim21 and small enough to be inserted into the plasmid.
−
 
+
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.
−
 
+
3XGS is the improvement we made this year, based on a 2XGS 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.
  
 
===Source===
 
===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.
−
ddddd
+
<br/>
 +
2-The sequence of VL&VH of erbB3 scFv comes from a patent: NO.US 9,346,889 B2
 +
<br/>
 +
3-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===
 
===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. <br/>
 +
[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.

Latest revision as of 02:07, 18 October 2018


erbB3-composite


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2574
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3243
    Illegal BamHI site found at 3781
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2531
    Illegal AgeI site found at 1376
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1886
    Illegal BsaI site found at 3621
    Illegal SapI.rc site found at 2234


Design Notes

Choosing nanobody as the ideal antibody in our project aims to avoid the disadvantage of traditional antibodies, among which the most difficult is the large sequence amount of these antibodies that may influence the transfection or the expression of the part. Linking the recombinate scFv of erbB3 and the hIgG1-Fc makes a promising recombinant antibody that is both recognizable by trim21 and small enough to be inserted into the plasmid. 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. 3XGS is the improvement we made this year, based on a 2XGS 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.

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 VL&VH of erbB3 scFv comes from a patent: NO.US 9,346,889 B2
3-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.