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Fig. 8. Localization of Mdy2. (A) Vegetatively growing cells. The mutant mdy2 (HZH686) was transformed with a plasmid encoding a functional GFP-Mdy2 fusion protein or with the empty vector as a control. Expression of the fusion gene was induced by incubating the cells in 3% raffinose and 1% galactose, and analysed by fluorescence and phase-contrast microscopy. (B) Induction with pheromone. mdy2 mutant cells were transformed with plasmids expressing a functional GFP-Mdy2 fusion protein or with the empty vector as a control. Cells were induced as above, then treated with 5 µM -factor for 2 hours, and analysed by fluorescence and phase-contrast microscopy. (C) Expression of GFP-MDY2 under its own promoter. Localization of GFP-Mdy2 expressed under the control of its own promoter. Cultures of the mdy2 mutant (HZH686) harbouring MDY2p-GFP-MDY2 (pZH152) were grown to log phase. Half of the culture was then harvested and the localization of GFP-Mdy2 fusion protein was investigated. (D) The other half was treated with 5 µM -factor for 2 hours before fusion protein localization was investigated.

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