Use of copper sulphate and Paraquat as selectable resistance markers for stable maintenance of yeast recombinant plasmids

• Published in: Biotechnology letters Vol. 19; no. 11; pp. 1103 - 1107
• Main Authors: Merlotti, M.C, Rech, S.B, Oliver, S.G
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.11.1997; Springer Nature B.V
• Subjects: Biological and medical sciences; Biotechnology; Fermentation; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; genetic transformation; Methods. Procedures. Technologies; Microbiology; Miscellaneous; Paraquat; plasmid vectors; Plasmids; Synthetic digonucleotides and genes. Sequencing; yeast episomal plasmids; Yeasts; Recombinant microorganism; Yeast; Stability; Genetic transformation; Selection; Herbicide; Fungi; Plasmid; Resistance; Copper Sulfates; Paraquat; Ascomycetes; Saccharomyces cerevisiae; Thallophyta
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1023/A:1018440626941

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.