Difference between revisions of "Part:BBa K2653001"
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===Design Notes===
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===Usage and Biology===
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The GFP-nano works as a core in our system to degrade the proof of concept target GFP as the tool to bind with it. In the design we link GFP-nano with hIgG1-Fc, so that it can work as the bridge between target GFP and Trim21, and finally help form a ternary complex of GFP, GFP-nano and Trim21. Only after this the degradation could successfully be processed.
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The GFP-nano works as a core in our system to degrade the proof of concept target GFP as the tool to bind with it. In the design we link GFP-nano with hIgG1-Fc, so that it can work as the bridge between target GFP and Trim21, and finally help form a ternary complex of GFP, GFP-nano and Trim21. Then the E3 ubiquitin ligase Trim21 would tag the complex with the ubiquitin mark, and then employ the proteasome-ubiquitin system to degrade the target GFP protein.
  
 
===Source===
 
===Source===

Revision as of 12:05, 17 October 2018


GFP-nano

The antibody GFP-nano is designed to bind with both Trim21 and the antigen GFP to prove that the P.R.PREDATOR really works. It belongs to the single-chain variable fragment(scFv) family, which has the ability to bind with GFP, but is much smaller than the common antibody of GFP(only 364 bps). In the project GFP-nano successfully bound with target GFP.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 35


Usage and Biology

The GFP-nano works as a core in our system to degrade the proof of concept target GFP as the tool to bind with it. In the design we link GFP-nano with hIgG1-Fc, so that it can work as the bridge between target GFP and Trim21, and finally help form a ternary complex of GFP, GFP-nano and Trim21. Then the E3 ubiquitin ligase Trim21 would tag the complex with the ubiquitin mark, and then employ the proteasome-ubiquitin system to degrade the target GFP protein.

Source

The sequence of GFP-nano we use came from a patent.

References

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