Applied Microbiology and Biotechnology

, Volume 30, Issue 4, pp 388–394 | Cite as

G418 resistance in the yeast Saccharomyces cerevisiae: comparison of the neomycin resistance genes from Tn5 and Tn903

  • Christine Lang-Hinrichs
  • Dietmar Berndorff
  • Carsten Seefeldt
  • Ulf Stahl
Applied Genetics and Regulation

Summary

The neo genes of Tn5 and Tn903 (Tn601) coding for amigoglycoside phosphotransferase type II and type I, respectively, were joined to the yeast adc 1 promoter and trp1 terminator and introduced into yeast (Saccharomyces cerevisiae) cells. Transformants were obtained by direct selection for G418 resistance. Plasmids containing the Tn5 neo gene induced antibiotic resistance only at low frequency, whereas colonies transformed with the Tn903 neo gene could be selected at high frequency (300–400 transformants/μg plasmid DNA). The resistance threshold of transformed strains was increased to 30 mg G418/ml by both genes and high level expression of the bacterial genes in yeast could be shown using an in vitro phosphotransferase assay. The results indicate that this system can be used for high frequency transformation of wild-type strains and might in addition be used for the identification and isolation of promoter-active sequences.

Abbreviations

adc 1

alcoholdehydrogenase I gene

APH

aminoglycoside-3′-phosphotransferase

leu2

β-isopropylmalate dehydrogenase gene

neo

aminoglycoside-3′-phosphotransferase gene

trp1

N-(5′-phosphoribosyl)-anthranilate isomerase gene

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Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Christine Lang-Hinrichs
    • 1
  • Dietmar Berndorff
    • 2
  • Carsten Seefeldt
    • 2
  • Ulf Stahl
    • 2
  1. 1.Institut für Gärungsgewerbe und BiotechnologieBerlin 65
  2. 2.Fachgebiet MikrobiologieTechnische Universität Berlin Fachbereich 132Berlin 65

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