FIGURE 6: Different mitochondrial targeting signals direct proteins reliably to the different mitochondrial subcompartments. (A) Schematic representation of the different targeting signals and proteins used for fusions to sfGFP_1-10 and sfGFP₁₁. (B) Schematic representation of the split GFP constructs used. The plasmids were created using the cHHYTK15 plasmid as described in the Materials and Methods section. The expression of proteins from the RNR1 promoter is comparable to that from the RNR2 promoter [14]. (C) YPH499 wild type cells were expressed with the respective plasmid pairs, grown in glucose containing medium to mid-log phase and analyzed by fluorescence microscopy. Representative images are shown. Scale bar, 5 µm. (D) The fluorescence in the different cells was measured by fluorescence spectroscopy in a microtiter plate reader. The maximum intensity of constructs 1-5 were set to 1 (except for the Ubc6-TA sample for which only background fluorescence was measured). The correct matching combinations are indicated by red frames and yellow check marks.

14. Lee ME, DeLoache WC, Cervantes B, Dueber JE (2015). A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly. ACS Synth Biol 4(9): 975-986. 10.1021/sb500366v