Table 6.
Strains and markers used in Y1H and Y2H assays
| Y1H | Y2H | |
|---|---|---|
| Bait | DNA | Protein |
| Bait construct | Integrated | Plasmid |
| Bait marker | HIS3, URA3 | LEU2 |
| Bait strain | YM4271 (mating type a) | MaV103 (mating type a) |
| Prey | Protein | Protein |
| Prey construct | Plasmid | Plasmid |
| Prey marker | TRP1 | TRP1 |
| Prey straina | Yα1867 (mating type α) | MaV203 (mating type α) |
| Reporter | HIS3 (with 3AT), LacZ | HIS3 (with 3AT), LacZ, URA3 |
| Media used in screen | Sc–Ura–His–Trp+3AT | Sc–Leu–Trp–His+3AT |
| Counter selection | No | Yes, with 5FOA |
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Y1H, yeast one-hybrid; Y2H, yeast two-hybrid.
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aOnly relevant when assay is performed by mating rather than by transformation (Vermeirssen et al. 2007b) (see Protocol: Identifying Interactors from an Activation Domain Prey Library [Reece-Hoyes and Walhout 2018d]).
