Use of copper sulphate and Paraquat as selectable resistance markers for stable maintenance of yeast recombinant plasmids
1997
Merlotti, M.C. | Rech, S.B. | Oliver, S.G.
Saccharomyces cerevisiae SOD2 and CUP1 genes were used to maintain high-copy number plasmids (YEp) in laboratory and industrial yeast strains. The plasmid, YEpS1, containing the SOD2 gene was unstable in a sod2 mutant. However when Paraquat (0.5 mM) was used as a selective agent, the plasmid was maintained in the sad2 mutant but lost in the wild-type strain. When the CUPi gene was inserted into YEpS1, the resulting plasmid (YEpCuS1) was 100% stable in the sod2 mutant grown in Cu2-containing medium. In the absence of Cu2+, the proportion of plasmid-containing cells fell to 20%. YEpS1 was also transformed into an industrial strain, transformants could be selected in Paraquat-containing medium but showed poor stability.
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