Team:Tsinghua/Chart
Part Charts
Basic Parts
| Name | Short description | Long description |
|---|---|---|
| EBBa K2980000 | Cry2 | Cryptochrome 2 (CRY2) is a blue light stimulated photoreceptor, when exposed to blue light, it would interact with CIB1 (Part:BBa K2980002). Technically, we use 488nm laser of confocal microscope, which also activate GFP, to stimulate the binding of two light-control element. Functionally, it is used to fuse with other protein and bring them together into phase under light. |
| BBa K2980001 | mCherry | Monomeric derivative of DsRed fluorescent protein. It is used as a reporter in fusion protein to indicate the location of fusion protein in cell. |
| BBa K2980002 | CIB1 | CIB1 will interact with CRY2 (Part:BBa K2980000) under blue light stimulation. It is the main light control element in our system. |
| BBa K2980003 | SUMO | Ubiquitin-like protein that can be covalently attached to proteins as a monomer, and it can interact with domain SIM (Part:BBa K2980004). |
| BBa K2980004 | SIM | SUMO binding domain, which can bind SUMO (Part:BBa_K2980003) in vivo. |
| BBa K2980005 | FUSLCD | It is the low complexity region of FUS, a RNA binding protein that perform various functions during transcription initiation, RNA splicing and transportation. When this protein accumulate in cytoplasm, it could phase separate and influence RNA metabolism. |
| BBa K2980007 | GCN(4) | Naturally, it is a transcription factor that is responsible for the activation of more than 30 genes. It can interact with the same domain as itself. It is used as oligomerizing tag of phase-separation element. |
| BBa K2980008 | mEGFP | Enhanced GFP with monomerizing A206K mutation. It is used as a reporter in fusion protein to indicate the location of fusion protein in cell. |
| BBa K2980011 | CRY2(R489E) | Cry2 (R489E) is a mutant of Cryptochrome 2 (Cry2, Part:BBa_K1982009). By changing arginine on 489 to glutamate, the charge of C terminal is changed and the ability of CRY2 homo-oligomerization is lowered. |
Composite Parts
| Name | Short description | Long description |
|---|---|---|
| EBBa K2980006 | Cry2-mCherry | A fusion protein that is component of phase-separation system. When stimulated by blue light, it will be recruited into the phase formed by CIB1-GCN(4)-mEGFP-FUSLCD (Part:BBa K2980009). |
| BBa K2980009 | CIB1-GCN(4)-mEGFP-FUSLCD | The fusion protein can induce consistant phase-separation in cell once well expressed. Under blue light stimulation, cry2-mcherry (Part:BBa K2980006) will be recruited into its phase. |
| BBa K2980010 | Cry2-mCherry-xylE | Cry2-mCherry-xylE with Anderson promoter and LacI binding regulator region. We use this to prove the effect of phase separation on enzyme catalysis. |
| BBa K2980012 | Cry2-mCherry-Rluc | Rluc (Part:BBa_K1722005) was linked to the C terminal of Cry2-mCherry (Part:BBa_K2980006) and co-transformed with CIB1-GCN(4)-mEGFP-FUSLCD （Part:BBa_K2980009) to make it possible for Rluc to be recruited into phase. |
| BBa K2980013 | lacI-GFP-FUS | This part can induce asymmetric cell division in E.coli. FUSLCD in this part would aggregate at one end of cells when expressing at proper level. At the same time, lacI could attach target plasmids with lacO on it, recruiting it to the end of cell with FUSLCD. As the plasmids attracted to an end, the two filial cells would differ in the copy number of target plasmid. |
| BBa K2980014 | (SIM-SUMO)3-SUMO3-mEGFP-Cry2 | This part is the fusion of a photo-regulated protein Cry2, a green fluorescence protein mEGFP, and tandem repeats of SIM-SUMO as well as SIM. This part undergoes liquid-liquid phase separation in E. coli, forming green spherical liquid-like puncta. It can be used for light-induced recruitment of other clients into phase. |
| BBa K2980015 | CIB1-mCherry | A fusion protein that is component of phase-separation system. When stimulated by blue light, it will be recruited into the phase formed by (SIMSUMO)3-SUMO3-GFP-Cry2 (Part:BBa K2980014). |
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