PICT
PICT (Protein interactions from Imaging Complexes after Translocation) is a method that allows the accurate measurement of parameters that define protein complexes in a physiological context. This technique provides a simple fluorescence microscopy readout for the study of protein-protein interactions and their dynamics in living cells.
Protocol
A step-by-step protocol for your own PICT experiment!
Software PyF2F
Find here our repository in GitHub, PyF2F and more!
Reagents: Strains
| Strain | Genotype | Source |
|---|---|---|
| OGY0007 | MATα, his3Δ1, leu2Δ0, ura3Δ0, LYS2+, can1Δ::STE2pr-LEU2, Lyp1Δ::, tor1-1 | Gallego et al. 2013, PlosOne |
| OGY0008 | MATα, his3Δ1, leu2Δ0, ura3Δ0, LYS2+, can1Δ::STE2pr-LEU2, Lyp1Δ::, tor1-1, fpr1Δ::klUra | Gallego et al. 2013, PlosOne |
| OGY0009 | MATα, his3Δ1, leu2Δ0, ura3Δ0, LYS2+, can1Δ::STE2pr-LEU2, Lyp1Δ::, tor1-1, fpr1Δ::kanMX4 | Picco et al. 2017, Cell |
| OGY0010 | MATα, his3Δ1, leu2Δ0, ura3Δ0, LYS2+, can1Δ::STE2pr-LEU2, Lyp1Δ::, tor1-1, fpr1Δ::klUra, SLA2-RFP-FKBP::natNT2 | Picco et al. 2017, Cell |
| OGY0012 | MATα, his3Δ1, leu2Δ0, ura3Δ0, LYS2+, can1Δ::STE2pr-LEU2, Lyp1Δ::, tor1-1, fpr1Δ::klUra, PIL1-RFP-FKBP::natNT2 | Gallego et al. 2013, PlosOne |
| OGY0015 | MATa, his3Δ1, leu2Δ0, met15Δ0, ura3Δ0, SLA2-RFP-FKBP::natNT2 | Picco et al. 2017, Cell |
| OGY0336 | MATa, his3-200, leu2-3,112, ura3-52, lys2-801, tor1-1, fpr1::, TUB4-RFP-FKBP-natNT2 | Torreira et al. 2017, eLife |
| OGY0607 | MATa, his3Δ1, leu2Δ0::HIS3MX6-GAL1pr-I-SCEI, met15Δ0, ura3Δ0, SLA2-RFP-FKBP::natNT2 | Picco et al. 2017, Cell |
| OGY0992 | MATa, his3Δ1, leu2Δ0, ura3Δ0, LYS2+, can1Δ::STE2pr-LEU2, lyp1Δ::, tor1-1, fpr1Δ::klURA, SLA2-RFP-FKBP::natNT2, EXO70-FRB::hphNT1, SEC5-3xmyeGFP::kanMX4 | Hernandez et al. 2024, NARGAB |
| OGY1322 | MATa, his3Δ1, leu2Δ0, ura3Δ0, LYS2+, can1Δ::STE2pr-LEU2, Lyp1Δ::, tor1-1, fpr1Δ::klUra, SLA2-RFP-FKBP::natNT2, EXO70-FRB::hphNT1, SEC5-mNeonGreen::kanMX4 | Hernandez et al. 2024, NARGAB |
| OGY1323 | MATa, his3Δ1, leu2Δ0, ura3Δ0, LYS2+, can1Δ::STE2pr-LEU2, Lyp1Δ::, tor1-1, fpr1Δ::klUra, SLA2-RFP-FKBP::natNT2, EXO70-FRB::hphNT1, SEC2-mNG::kanMX4 | Hernandez et al. 2024, bioRxiv |
Reagents: Plasmids
| Plasmid | Tag | Linker (amino acids) | Marker | Source |
|---|---|---|---|---|
| p0G010 | FRB(T2098L) | 12 | HphNT1 | Gallego et al. 2013, PlosOne |
| pOG012 | FRB(T2098L)-meGFP | 12 | KanMX4 | Gallego et al. 2013, PlosOne |
| pOG019 | RFP-FKBP | 24 | NatNT2 | Gallego et al. 2013, PlosOne |
| pOG043 | Gal1pr.I-ISCEI | His3MX6 | Picco et al. 2017, Cell |
