TABLE 3.
Two-hybrid reporter gene assays
| Plasmid combination
|
Reporter gene assay resulta
|
||
|---|---|---|---|
| Bait domain fusion | Activation domain fusion | SD-Leu-Trp-His | β-Galactosidase |
| pBD-Z1 | pAD-RpoE1 | ++ | + (5 h) |
| p53 | pAD-RpoE1 | + | − |
| pBD-Z2 | pAD-RpoE1 | + | − |
| pBD-E | pAD-RpoE1 | + | − |
| pBD-AsgA | pAD-RpoE1 | + | + (7 h) |
| p53 | pSV40 | +++++ | + (4 h) |
| pLaminC | pSV40 | + | − |
a
Various bait-domain fusions were tested against the activation domain-RpoE1 fusion for reporter gene activation in yeast. Expression of the HIS3 reporter gene was shown to be leaky under all conditions (+). Expression of β-galactosidase is reported as + for cells which turned blue on the filter paper and − for cells which showed no color change. The time when blue color was first observed on the filters is given in parentheses. The combination p53-pSV40 was used as the system positive control, while the combination pLaminC-pSV40 was used as the negative control.