Schematic diagram
BBa_K2601010
We combined Frb, yEGFP and HOTag6 togethor in BBa_K2601010. Frb can interact with FKBP after adding rapamycin, which makes the RapaSPOT formation controllable. We constructed yeast strains with FKBP-mCherry-HOTag3 and Frb-yEGFP-HOTag6 and used 10 μM rapamycin to induce SPOT formation. Minutes after adding rapamycin, granules appeared and became larger gradually.
BBa_K2601011
![]( https://static.igem.org/mediawiki/2018/8/87/T--Peking--parts2.jpeg)
Super Uranyl-binding Protein (SUP) was fused to 4*(VPGVG) spaced triple SpyTag. The fused protein is used to generate the uranyl-binding capacity in our polymer network system. Paired with triple SpyCatcher (BBa_K1989001) and triple SpyTag with mSA (BBa_K1989002), our polymer network system is constructed.
BBa_K2601012
Triple SpyCatcher was spaced by 15*(VPGVG). Paired with triple SpyTag with SUP (BBa_K1989000) and triple SpyTag with mSA (BBa_K1989002), our polymer network system is constructed.
BBa_ | Part |
K2601000 | Promoter-tet07 |
K2601001 | Promoter-PDH3 |
K2601002 | SUMO |
K2601003 | SIM |
K2601004 | HOTag3 |
K2601005 | HOTag6 |
K2601007 | Frb-yEGFP |
K2601008 | FKBP-yEGFP |
K2601010 | Frb-yEGFP-HOTag6 |
K2601011 | FKBP-yEGFP-HOTag3 |
K2601012 | SUMO-yEGFP-HOtag3 |
K2601021 | Tet07-Frb-yEGFP |
K2601023 | PDH3-Frb-yEGFP |
K2601025 | Tet07-FKBP-yEGFP |
K2601026 | TEF1-FKBP-yEGFP |
K2601027 | PDH3-FKBP-yEGFP |
K2601032 | Tet07-Frb-yEGFP-HOTag6 |
K2601033 | TEF1-Frb-yEGFP-HOTag6 |
K2601034 | PDH3-Frb-yEGFP-HOTag6 |
K2601037 | TEF1-FKBP-yEGFP-HOTag3 |
K2601038 | PDH3-FKBP-yEGFP-HOTag3 |
K2601040 | Tet07-SUMO-yEGFP-HOtag3 |
K2601041 | TEF1-SUMO-yEGFP-HOtag3 |
K2601042 | PDH3-SUMO-yEGFP-HOtag3 |
K2601054 | PDH3-Frb-crtE-HOTag6 |
K2601056 | Tet07-Frb-crtYB-HOTag6 |
K2601058 | PDH3-Frb-crtYB-HOTag6 |
K2601060 | SIM-crtE-Hotag6 |
K2601061 | SIM-crtYB-Hotag6 |
2018 Peking iGEM team members devoted themselves to constructing a part collection that can drive phase separation. We have not only submitted all of new basic HOTag parts, but also provided multiple composite parts combining HOTags with other protein-protein interaction modules and fluorescent reporters. The basic part, HOTag3, is a homo-oligomeric short peptide containing only 30 amino acids. It has high stoichiometry, forming hexamer spontaneously. The HOTag3, together with another tetrameric HOTag6, can robustly drive phase separation upon protein-protein interaction. Protein-protein interaction is achieved by our dimerization parts, including FKBP/Frb and SUMO/SIM. Functions of all the parts were thoroughly tested. Some thermodynamic and kinetic properties of the parts were characterized as well. We believe the HOTag is useful tool for other iGEM teams to investigate protein phase separation and design synthetic organelles.