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

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 mai...

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Bibliographic Details
Published in:Biotechnology letters 1997-11, Vol.19 (11), p.1103-1107
Main Authors: Merlotti, M.C, Rech, S.B, Oliver, S.G
Format: Article
Language:English
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
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Summary: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.
ISSN:0141-5492
1573-6776
DOI:10.1023/A:1018440626941