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. 2004 Jul;135(3):1457–1470. doi: 10.1104/pp.104.042820

Figure 2.

Figure 2.

Yeast two-hybrid interaction of HsfA2 in bait position with sHsps in prey position. For the bait construct, the C-terminal domain of LpHsfA2 was mutated in its two activator motifs (Döring et al., 2000). The residual growth of strains with the Gal4DBDxHsfA2.11.16B bait was abolished by adding 1 or 10 mm 3-amino-triazol (3-AT). In the prey position, we tested the indicated sHsps fused to the Gal4AD. Samples 4 to 9 represent tests with mutant forms of the two LpHsp17-CII isoforms (for sequence details see Fig. 1). In all cases, liquid cultures of the indicated yeast strains were grown overnight at 30°C in selective minimal medium without Trp and Leu (SD-WL). After adjustment to an OD600 of 0.1, 5 μL of these cultures as well as 2 subsequent dilutions by 1 to 10 were dropped on agar plates of SD-WL medium lacking His (SD-WLH). The His-auxotroph colony growth was monitored after incubation for 48 h at 30°C.