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Plasmid DNA transformation of yeast cells treated with chelating agents. The role of lipid peroxidation in inducing competence in yeast
- Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR
It is found that the pretreatment of yeast Saccharomyces cerevisiae (strains 746 and LL-20) cells with the chelating agents of transition metals (8-hydroxyquinoline, 1,10-phe-nanthroline) causes the appearance of competence for plasmid DNA transformation. The factors affecting the efficiency of yeast cell transformation are studied using 8-hydroxyquinoline and double-replicon plasmid RB4. Transformant yield under application of such a method reached 4×103-2×104 transformations per 10 u.g of plasmid DNA. The chelating agents are shown to increase lipid peroxide formation in yeast. The addition of the lipid oxidation inhibitor (ionol, the synthetic antioxidant) during cell treatment with 8-hydroxyquinoline decreases the transformant yield. The data obtained indicate that the effect of chelators on the yeast competence for transformation may be due to their influence on the membrane lipid peroxidation.
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