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. 1985 Dec;82(23):7909–7913. doi: 10.1073/pnas.82.23.7909

Construction of strains of Saccharomyces cerevisiae that grow on lactose.

K Sreekrishna, R C Dickson
PMCID: PMC390879  PMID: 3934664

Abstract

We have constructed strains of Saccharomyces cerevisiae that grow on lactose (Lac+). S. cerevisiae strain YNN27, which, like all S. cerevisiae, is unable to grow on lactose, was transformed with pKR1B-LAC4-1. This plasmid has a selectable marker gene conferring resistance to the antibiotic G418 and carries a 13-kilobase region of the Kluyveromyces lactis genome including LAC4, a beta-galactosidase gene. Transformants were selected first for G418 resistance and then for growth on lactose. Southern hybridization experiments showed that Lac+ transformants had integrated 15-25 tandem copies of the vector into a host chromosome. Several lines of evidence indicate that the Lac+ phenotype in pKR1B-LAC4-1-transformed S. cerevisiae is due to expression of a K. lactis lactose permease gene that lies between 2 and 8.6 kilobase upstream of LAC4 and also to expression of LAC4. The permease gene has been designated LAC12.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Dickson R. C., Barr K. Characterization of lactose transport in Kluyveromyces lactis. J Bacteriol. 1983 Jun;154(3):1245–1251. doi: 10.1128/jb.154.3.1245-1251.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dickson R. C. Expression of a foreign eukaryotic gene in Saccharomyces cerevisiae: beta-galactosidase from Kluyveromyces lactis. Gene. 1980 Sep;10(4):347–356. doi: 10.1016/0378-1119(80)90155-9. [DOI] [PubMed] [Google Scholar]
  3. Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hsiao C. L., Carbon J. High-frequency transformation of yeast by plasmids containing the cloned yeast ARG4 gene. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3829–3833. doi: 10.1073/pnas.76.8.3829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Johnston S. A., Hopper J. E. Isolation of the yeast regulatory gene GAL4 and analysis of its dosage effects on the galactose/melibiose regulon. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6971–6975. doi: 10.1073/pnas.79.22.6971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Laughon A., Gesteland R. F. Isolation and preliminary characterization of the GAL4 gene, a positive regulator of transcription in yeast. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6827–6831. doi: 10.1073/pnas.79.22.6827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Needleman R. B., Kaback D. B., Dubin R. A., Perkins E. L., Rosenberg N. G., Sutherland K. A., Forrest D. B., Michels C. A. MAL6 of Saccharomyces: a complex genetic locus containing three genes required for maltose fermentation. Proc Natl Acad Sci U S A. 1984 May;81(9):2811–2815. doi: 10.1073/pnas.81.9.2811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Serrano R. Energy requirements for maltose transport in yeast. Eur J Biochem. 1977 Oct 17;80(1):97–102. doi: 10.1111/j.1432-1033.1977.tb11861.x. [DOI] [PubMed] [Google Scholar]
  9. Sheetz R. M., Dickson R. C. Lac4 is the structural gene for beta-galactosidase in Kluyveromyces lactis. Genetics. 1981 Aug;98(4):729–745. doi: 10.1093/genetics/98.4.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Sreekrishna K., Webster T. D., Dickson R. C. Transformation of Kluyveromyces lactis with the kanamycin (G418) resistance gene of Tn903. Gene. 1984 Apr;28(1):73–81. doi: 10.1016/0378-1119(84)90089-1. [DOI] [PubMed] [Google Scholar]
  11. Stinchcomb D. T., Thomas M., Kelly J., Selker E., Davis R. W. Eukaryotic DNA segments capable of autonomous replication in yeast. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4559–4563. doi: 10.1073/pnas.77.8.4559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Terrell S. L., Bernard A., Bailey R. B. Ethanol from Whey: Continuous Fermentation with a Catabolite Repression-Resistant Saccharomyces cerevisiae Mutant. Appl Environ Microbiol. 1984 Sep;48(3):577–580. doi: 10.1128/aem.48.3.577-580.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Webster T. D., Dickson R. C. Direct selection of Saccharomyces cerevisiae resistant to the antibiotic G418 following transformation with a DNA vector carrying the kanamycin-resistance gene of Tn903. Gene. 1983 Dec;26(2-3):243–252. doi: 10.1016/0378-1119(83)90194-4. [DOI] [PubMed] [Google Scholar]

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