Back to article: Yeast can express and assemble bacterial secretins in the mitochondrial outer membrane

FIGURE 1: Moderate expression of secretins is not toxic to yeast cells. (A) Schematic representation of the PulD and InvG constructs used in this study. The numbering of the amino acid residues starts at the start codon and includes the processed signal sequence. Currently, there is no information on the various domains of SsaC. (B) Wild type yeast cells were transformed with a plasmid encoding the indicated bacterial secretins under the control of the inducible GAL promoter. Cells were grown in synthetic glucose-containing (S-Glu) medium to an OD600 of 1.0 and spotted in a 1:5 dilution series on synthetic medium plates containing Glucose (S-Glu), Galactose (S-Gal), or Galactose + 0.1% Glucose (SGal + 0.1% Glu). Plates were then incubated at the indicated temperatures. Two colonies for each strain were analysed. (C) Wild type yeast cells transformed with a plasmid encoding the indicated secretins were grown in the indicated liquid media until logarithmic phase and then lysed. The cell lysates were analysed by SDS-PAGE and immunodecoration with antibodies against PulD or the HA-tag. The cytosolic protein Bmh1 was used as a loading control.

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