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Fig. 1. Recombinant human 14-3-3 proteins complement the lethal disruption of yeast 14-3-3 genes. (A) GG1306-derived yeast strains expressing the indicated 14-3-3 isoforms were transferred from glucose medium (left) to 5-FOA-containing medium (right). Strains can only grow on this medium when they are able to lose plasmid YCplac33[BMH2], illustrating the ability of the foreign 14-3-3 isoform to take over the function of the yeast 14-3-3 proteins. (B) The absence of plasmid Ycplac33[BMH2] after 5-FOA treatment was determined by PCR. This plasmid yields a 400 bp product, besides a 600 bp internal control product. Western blots on 5 µg (C) or 2.5 µg (D) total protein from strains after 5-FOA treatment with an antiserum against yeast 14-3-3 proteins (C) or against Pan 14-3-3 proteins (D). Lane b contains 20 ng purified Bmh2p.





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