Mitochondria-targeted green fluorescent proteins: convenient tools for thestudy of organelle biogenesis in Saccharomyces cerevisiae

We describe the construction and characterization of a novel set of plasmid s for expression of mitochondria-targeted green fluorescent protein (GFP) i n Saccharomyces cerevisiae. The vectors include constructs with strong regu latable and constitutive promoters, four different auxotrophic markers for yeast transformation, and a green (S65T) and a blueshifted (P4-3) variant o f GFP, Mitochondria are brightly fluorescent in living yeast cells grown on different carbon sources and at different temperatures,,vith virtually no background staining. Specific staining of mitochondria is also shown for a respiratory-deficient mutant with abnormal mitochondrial morphology. The pl asmids facilitate convenient analysis of mutants defective in mitochondrial morphology or inheritance and, at the same time, are suitable vectors for easy construction of different kinds of GFP fusion proteins to study variou s aspects of organelle biogenesis in living yeast cells. Copyright (C) 2000 John Wiley & Sons, Ltd.