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. 2006 Feb;5(2):262–271. doi: 10.1128/EC.5.2.262-271.2006

FIG. 3.

FIG. 3.

Expression profiles of ATG mutants of Dhal2p in S. cerevisiae. (A) Complementation of methionine auxotrophy exhibited by RS1051 harboring plasmid pMA7 (Dhal2p), pMA23 (Dhal2p M3 ATG mutant), or pMA24 (Dhal2p M1/M2 ATG mutant). (B) Immunoblot showing expression of ATG mutants of Dhal2p in S. cerevisiae. Total protein extract (20 μg) from strain RS1051 harboring plasmid pMA23 or pMA24 was subjected to immunoblot analysis with anti-Dhal2p antibody. For comparison, protein extracts from cells with plasmid pMA14 (expressing both full-length and truncated Dhal2p) and plasmid pMA15 (expressing only truncated Dhal2p) were loaded. Purified recombinant truncated Dhal2p is shown as Control. The blot shown is a representative of three different experiments. (C) Immunoblot showing localization of ATG mutants of Dhal2p in S. cerevisiae. Cytosolic (S) and membrane (P) fractions from strain RS1051 harboring plasmid pMA23 or pMA24 were subjected to immunoblot analysis using anti-Dhal2p antibody. The experiment was repeated three times with similar results.