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Recombinant Gene Expression Protocols pp 149–157Cite as

Inducible Expression Cassettes in Yeast: ADH2

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 62))

Abstract

The ADH2 promoter belongs to a class of promoters subject to catabolite repression. Many of these promoters have strong transcriptional start signals, which are repressed by the presence of glucose. The strength of the ADH2 promoter coupled with the fact that the promoter is tightly regulated make it highly amenable for use in the expression of heterologous genes.

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References

  1. Denis, C. L. and Young, E. T. (1983) Isolation and characterization of the positive regulatory gene ADR1 from Saccharomyces cerevisiae. Mol. Cell Biol. 3, 360–310.

    PubMed  CAS  Google Scholar 

  2. Denis, C. L., Ciriacy, M., and Young, E. T. (1981) A positive regulatory gene is required for accumulation of the functional messenger RNA for the glucose-repressible alcohol dehydrogenase from Saccharomyces cerevisiae. J. Mol. Biol. 148, 355–368.

    Article  PubMed  CAS  Google Scholar 

  3. Ciriacy, M. (1975) Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. II Two loci controlling synthesis of the glucose-repressible ADHII. Mol. Gen. Genet. 138, 157–164.

    Article  PubMed  CAS  Google Scholar 

  4. Ciriacy, M. (1977) Isolation and characterization of mutants defective in intermediary carbon metabolism and in carbon catabolite repression. Mol. Gen. Genet. 154, 213–220.

    Article  PubMed  CAS  Google Scholar 

  5. Denis, C. L. (1984) Identification of new genes involved in the regulation of yeast alcohol dehydrogenase II. Genetics 108, 833–844.

    PubMed  CAS  Google Scholar 

  6. Yu, J., Donoviel, M. S., and Young, E. T. (1989) Adjacent upstream activation sequence elements synergistically regulate transcription of ADH2 in Saccharomyces cerevisiae. Mol. Cell Biol. 9, 34–42.

    PubMed  CAS  Google Scholar 

  7. Eisen, A., Taylor, W. E., Blumberg, H., and Young, E. T. (1988) The yeast regulatory protein ADR1 binds in a zinc-dependent manner to the upstream activating sequence of ADH2. Mol. Cell Biol. 8, 4552–4556.

    PubMed  CAS  Google Scholar 

  8. Beier, D. R. and Young, E. T. (1982) Characterization of a regulatory region upstream of the ADR2 locus of S. cerevisiae. Nature 300, 724–728.

    Article  PubMed  CAS  Google Scholar 

  9. Beier, D. R., Sledziewski, A., and Young, E. T. (1985) Deletion analysis identifies a region, upstream of the ADH2 gene of Saccharomyces cerevisiae, which is required for ADR1-mediated derepression. Mol. Cell Biol. 5, 1743–1749.

    PubMed  CAS  Google Scholar 

  10. Russell, D. W., Smith, M., Cox, D., Williamson, V. M., and Young, E. T. (1983) DNA sequences of two yeast promoter-up mutants. Nature 304, 652–654.

    Article  PubMed  CAS  Google Scholar 

  11. Ciriacy, M. (1979) Isolation and characterization of further cis-and transacting regulatory elements involved in the synthesis of glucose-repressible alcohol dehydrogenase (ADHII). Mol. Gen. Genet. 176, 427–431.

    Article  PubMed  CAS  Google Scholar 

  12. Thukral, S. K., Eisen, A., and Young, E. T. (1991) Two monomers of yeast transcription factor ADR1 bind a palindromic sequence symmetrically to activate ADH2 expression. Mol. Cell. Biol. 11, 1566–1577.

    PubMed  CAS  Google Scholar 

  13. Cook, W. J. and Denis, C. L. (1993) Identification of three genes required for the glucose-dependent transcription of the yeast transcriptional activator ADR1. Curr. Genet. 23, 192–200.

    Article  PubMed  CAS  Google Scholar 

  14. Denis, C. L. and Gallo, C. (1986) Constitutive RNA synthesis for the yeast activator ADR1 and identification of the ADR1-5 c mutation implications in posttranslational control of ADR1. Mol. Cell. Biol. 6, 4026–4030.

    PubMed  CAS  Google Scholar 

  15. Denis, C. L., Fontaine, S. C., Chase, D., Kemp, B. E., and Bemis, L. T. (1992) ADR1c mutations enhance the ability of ADR1 to activate transcription by a mechanism that is independent of effects on cyclic AMP-dependent protein kinase phosphorylation of Ser-230. Mol. Cell. Biol. 12, 1507–1514.

    PubMed  CAS  Google Scholar 

  16. Cook, W. J., Chase, D., Audino, D. C., and Denis, C. L. (1994) Dissection of the ADR1 protein reveals multiple, functionally redundant activation domains interspersed with inhibitory regions: evidence for a repressor binding to the ADR1c region. Mol. Cell. Biol. 14, 629–640.

    PubMed  CAS  Google Scholar 

  17. Denis, C. L. (1987) The effects of ADR1 and CCR1 gene dosage on the regulation of the glucose-repressible alcohol dehydrogenase from Saccharomyces cerevisiae. Mol. Gen. Genet. 208, 101–106.

    Article  PubMed  CAS  Google Scholar 

  18. Cherry, J. R. and Denis, C. L. (1989) Overexpression of the yeast transcriptional activator ADR1 induces mutation of the mitochondrial genome. Curr. Genet. 15, 311–317.

    Article  PubMed  CAS  Google Scholar 

  19. Price, V. L., Taylor, W. E., Clevenger, W., Worthington, M., and Young, E. T. (1990). Expression of heterologous proteins in Saccharomyces cerevisiae using the ADH2 promoter. Methods Enzymol. 185, 308–318.

    Article  PubMed  CAS  Google Scholar 

  20. Price, V., Mochizuki, D., March, C. J., Cosman, D., Deeley, M. C., Klinke, R., Clevenger, W., Gillis, S., Baker, P., and Urdal, D. (1987) Expression, purification and characterization of recombinant murine granulocyte-macrophage colony stimulating factor and bovine IL-2 from yeast. Gene 55, 287–293.

    Article  PubMed  CAS  Google Scholar 

  21. Kaslow, D. C., Hui, G., and Kumar, S. (1994) Expression and antigenicity of Plasmodium falciparum major merozoite surface protein (MSP19) variants secreted from Saccharomyces cerevisiae. Mol. Biochem. Parasitol. 63, 283–289.

    Article  PubMed  CAS  Google Scholar 

  22. Kaslow, D. C. and Shiloach, J. (1994) Production, purification and immunogenicity of a malaria transmission-blocking vaccine candidate TBV25H expressed in yeast and purified using nickel-NTA agarose. Biotechnology 12, 494–499.

    Article  PubMed  CAS  Google Scholar 

  23. Bishop, P. D., Teller, D. C., Smith, R. A., Lasser, G. W., Gilbert, T., and Seale, R. L. (1990) Expression, purification, and characterization of human factor XIII in Saccharomyces cerevisiae. Biochemistry, 29, 1861–1869.

    Article  PubMed  CAS  Google Scholar 

  24. Russell, D. W. and Smith, M. (1983) Nucleotide sequence of the yeast alcohol dehydrogenase II gene. J. Biol. Chem. 258, 2674–2682.

    PubMed  CAS  Google Scholar 

  25. Hill, J. E., Myers. A. M., Koerner, T. J., and Tzagoloff, A. (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2, 163–167.

    Article  PubMed  CAS  Google Scholar 

  26. Brunelli, J. P. and Pall, M. L. (1993) A series of yeast/Escherichia coli 1 expression vectors designed for directional cloning of cDNAs and cre/lox-mediated plasmid excision. Yeast 9, 1309–1318.

    Article  PubMed  CAS  Google Scholar 

  27. Sathe, G. M., O’Brien, S., McLaughlin, M. M., Watson, F., and Livi, G. P. (1991) Use of polymerase chain reaction for rapid detection of gene insertions in whole yeast cells. Nucleic Acids Res. 19, 4775.

    Article  PubMed  CAS  Google Scholar 

  28. Barnes, W. M. (1994) PCR amplification of up to 35-kb DNA with high fidelity and high yield from λ bacteriophage templates. Proc. Natl. Acad. Sci. USA 91, 2216–2220.

    Article  PubMed  CAS  Google Scholar 

  29. Gearing, D. P., Thut, C. J., VandenBos, T., Gimpel, S. D., Delaney, P. B., King, J., Price, V., Cosman, D., and Beckmann, M. P. (1991) Leukemia inhibitory factor receptor is structurally related to the IL-6_signal transducer, gp130. EMBO J. 10, 2839–2848.

    PubMed  CAS  Google Scholar 

  30. Curtis, B. M., Williams, D. E., Broxmeyer, H. E., Dunn, J., Farrah, T., Jeffery, E., Clevenger, W., deRoos, P., Martin, U., Friend, D., Craig, V., Gayle, R., Price, V., Cosman, D., March, C. J., and Park, L. S. (1991) Enhanced hematopoietic activity of a human granulocyte/macrophage colony-stimulating factor-interleukin 3 fusion protein. Proc. Natl. Acad. Sci. USA 88, 5809–5813.

    Article  PubMed  CAS  Google Scholar 

  31. Adams, S. E., Dawson, K. M., Gull, K., Kingsman, S. M., and Kingsman, A. J. (1987) The expression of hybrid HIV Ty virus-like particles in yeast. Nature 329, 68–70.

    Article  PubMed  CAS  Google Scholar 

  32. Chinery, S. A. and Hinchliffe, E. (1989) A novel class of vector for yeast transformation. Curr. Genet. 16, 21–25.

    Article  PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Immunex Corporation, Seattle, WA

    Virginia L. Price

Authors
  1. Virginia L. Price
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Editor information

Editors and Affiliations

  1. Thomas Jefferson University, Philadelphia, PA

    Rocky S. Tuan

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© 1997 Humana Press Inc.

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Cite this protocol

Price, V.L. (1997). Inducible Expression Cassettes in Yeast: ADH2. In: Tuan, R.S. (eds) Recombinant Gene Expression Protocols. Methods in Molecular Biology, vol 62. Humana Press. https://doi.org/10.1385/0-89603-480-1:149

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  • DOI: https://doi.org/10.1385/0-89603-480-1:149

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-480-8

  • Online ISBN: 978-1-59259-548-8

  • eBook Packages: Springer Protocols

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