Team:NUDT CHINA/Part Collection
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Part Collection
Introduction
This part collection, namely PPI Tool-kit, contained 18 new parts design for protein-protein interaction (PPI) researches.
In PPI Toolkit, we collected basic parts containing the coding sequence of three split-reporting systems: split-GFP, split-Luciferase, and split-HRP. All those basic parts were codon-optimized to be expressed in E.coli, and sequence-modified to meet the demands of RCF10. Building on that, amazing parts combining split-reporting system with a novel substitution system were designed, characterized and then added into the collection. Through which two proteins could be fused with their corresponding split-reporter fragment at the same time in one simple Golden-Gate Assembly, thus dramatically simplified the cloning process.
Figure 1. Schematic representation of the workflow of the substitution system.
All parts in this kit were functional validated, well characterized and submitted to the registry.
Parts in our Collection
N-sGFP/ C-sGFP based fluorescent PPI Detector
| BBa_K1997004 | sGFP-N->Zif268->sGFP-C |
| BBa_K1997015 | P+R->sGFP-N->Zif268->sGFP-C->TER |
| BBa_K1997016 | P+R->sGFP-N->Zif-N->RBS->Zif-C->sGFP-C->TER |
| BBa_K1997017 | P+R->sGFP-N->Zif-N->RBS->Zif-C->sGFP-C->TER |
GFP10/ GFP11+GFP1-9 based fluorescent PPI Detector
| BBa_K1997014 | GFP10->Zif268->GFP11->RBS->GFP1-9 |
| BBa_K1997018 | P+R->GFP10->Zif268->GFP11->RBS->GFP1-9->TER |
| BBa_K1997019 | P+R->GFP10->Zif-N->RBS->Zif-C->GFP11->RBS->GFP1-9->TER |
| BBa_K1997020 | P+R->GFP10->FRB->RBS->FKBP ->GFP11->RBS->GFP1-9-> TER |
Split-HRP based enzymatic PPI Detector
| BBa_K1997000 | sHRP-N |
| BBa_K1997001 | sHRP-C |
| BBa_K1997005 | sHRP-N->FRB->RBS->FKBP-> sHRP-C |
| BBa_K1997012 | P+R->sHRP-N->Zif268-> sHRP-C->TER |
| BBa_K1997013 | P+R->sHRP-N->Zif-N->RBS->Zif-C->sHRP-C->TER |
Split-Luciferase based chemiluminescence PPI Detector
| BBa_K1997002 | sLuc-N |
| BBa_K1997003 | sLuc-C |
| BBa_K1997021 | P+R->sLuc-N->Zif268->sLuc-C->TER |
| BBa_K1997022 | P+R->sLuc-N->Zif-N->RBS->Zif-C->sLuc-C->TER |
| BBa_K1997023 | P+R->sLuc-N->FRB->RBS->FKBP->sLuc-C->TER |
