Part:BBa_K3396005
Trim21-DocS
This composite part derives from TRIM21 and replace its PRYSPRY domain with DocS, and it perform the same function with the original TRIM21. What’s more, HA is added to the N-terminal of TRIM21, making it easier for TRIM21 to be detected by Western Blotting.
Usage and Biology
To demonstrate that the TRIM21 still works after replacing its PRYSPRY domain and keep its connection with all kinds of nanobodies, the PRYSPRY domain was replaced by DocS. Here is the mechanism of the recombined TRIM21-DocS:
1. The GFPnano tagged with Coh2 combines with targeted protein.
2. TRIM21-DocS connect Coh2-GFPnano-target through the DocS-Coh2 interaction.
3. The targeted protein is degraded by ubiquitin-proteasome system recruited by TRIM21.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 204
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 161
- 1000COMPATIBLE WITH RFC[1000]
===Improvement by team CU_Egypt 2022)
Improvement
There seemed to be some mistakes in the original sequence uploaded by NUDT 2020 (BBa_K3396005), there was an additional stop codon in the middle of the fusion protein and an extra Tyrosine residue in their Glycine-Serine Linker. These misconceptions hinder the usage of this part by any iGEM team, so we ought to provide them with the correct sequence that they could use easily if they want to assemble this part.
Figure 1.: Alignment of modified Trim-G4S-DocS sequence with the original sequence from of NUDT_2020 (BBa_K3396007) showing the extra amino acid residues (Valine-Leucine-Glutamic acid- Lysine) and the stop codon in the middle.
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