Difference between revisions of "Part:BBa K2653016:Design"
(→Source) |
(→References) |
||
| Line 19: | Line 19: | ||
===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. | + | [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. | [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
GFP-nano-IgG1-FC-HA TAG-Trim21
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2813
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3482
Illegal BamHI site found at 4020 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2770
Illegal AgeI site found at 1615 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 721
Illegal BsaI site found at 2125
Illegal BsaI site found at 3860
Illegal SapI.rc site found at 2473
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 nanobody of GFP 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 GFP-nano comes from a Korean patent:patent NO.10-2012-0049206.
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.