Table 1.
Interaction of p66 binding domain fusions with p51 activation domain fusions in the Y2H system
| Constructs | Operator | β-gal activity
|
|
|---|---|---|---|
| Colony* | Liquid† | ||
| p66SH2-1 + pGADNOT | lexA | − | ND |
| p66SH2-1 + pACTII | lexA | − | 0.02 |
| p66SH2-1 + p51GADNOT | lexA | ++ | 0.5 |
| p66SH2-1 + p51ACTII | lexA | +++ | 3.5 |
| p66AlaLex202 + pGADNOT | lexA | − | ND |
| p66AlaLex202 + pACTII | lexA | − | 0.04 |
| p66AlaLex202 + p51GADNOT | lexA | +++ | 1.6 |
| p66AlaLex202 + p51ACTII | lexA | +++ | 7.7 |
| p66NLexA + pGADNOT | lexA | − | 0.06 |
| p66NLexA + pACTII | lexA | − | 0.04 |
| p66NLexA + p51GADNOT | lexA | +++ | 6.6 |
| p66NLexA + p51ACTII | lexA | +++ | 25.0 |
| p66Lex202 + pGADNOT | lexA | +/− | ND |
| p66Lex202 + p51GADNOT | lexA | +/− | ND |
| p66GBT9 + pGADNOT‡ | UASG | − | ND |
| p66GBT9 + p51GADNOT‡ | UASG | − | ND |
Yeast strain CTY10-5d or ‡HF7c were transformed with plasmids encoding p66 bait and p51 prey fusions. Fusion proteins encoded by plasmids are described in Materials and Methods and Fig. 1.
*
Transformants were lifted onto nitrocellulose and subjected to the β-gal colony lift assay to determine intensities of blue color produced; +++, strong blue in 1 h; ++, blue in 1 h; +/−, weak blue in 3 h; −, white; ND, not done.
† Numbers represent β-gal activity in Miller units.