Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

Evaluation of Foreign Gene Codon Optimization in Yeast: Expression of a Mouse IG Kappa Chain

Bio/Technology volume 9, pages 1386–1389 (1991)Cite this article

Abstract

We have optimized the codons in an immunoglobulin kappa chain gene to those preferred in the yeast Saccharomyces cerevisiae. The mutant and wild type kappa chain genes were each fused with a synthetic invertase signal peptide that also contained only yeast–preferred codons, and expressed in the F762 yeast strain. The use of yeast–preferred codons resulted in a more than 5–fold increase in the rate of synthesis and at least a 50–fold increase in the steady state level of protein.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Skerra, A. and Plückthun, A. 1988. Assembly of a functional immunoglobulin Fv fragment in Escherichia coli. Science 240: 1038–1041.

    Article  CAS  Google Scholar 

  2. Plückthun, A. and Skerra, A. 1989. Expression of functional antibody Fv and Fab fragments in E. coli. Meth. in Enzymology 178: 497–515.

    Article  Google Scholar 

  3. Better, M. and Horwitz, A.H. 1989. Expression of engineered antibodies and antibody fragments in microorganisms. Methods in Enzymol. 178: 476–496.

    Article  CAS  Google Scholar 

  4. Hasemann, C.A. and Capra, J.D. 1990. High-level production of a functional immunoglobulin heterodimer in a baculovirus expression system. Proc. Nat. Acad. Sci. USA 87: 3942–3946.

    Article  CAS  Google Scholar 

  5. zu Putlitz, J., Kubasek, W.L., Duchene, M., Matthias, M., von Specht, B.-U. and Horst, D. 1990. Antibody production in baculovirus-infected insect cells. Bio/Technology 8: 651–654.

    CAS  PubMed  Google Scholar 

  6. Schekman, R. 1985. Protein localization and membrane traffic in yeast. Ann. Rev. Cell Biol. 1: 115–143.

    Article  CAS  Google Scholar 

  7. Zsebo, K.M., Lu, H.-S., Fieschko, J.C., Goldstein, L., Davis, J., Duker, K., Suggs, S.V., Lai, P.-H. and Bitter, G.A. 1986. Protein secretion from Saccharomyces cerevisiae directed by the prepro-α-factor leader region. J. Biol. Chem. 261: 5858–5865.

    CAS  PubMed  Google Scholar 

  8. Miyajima, A., Bond, M.W., Otsu, K., Arai, K.-i and Arai, N. 1985. Secretion of mature mouse interleukin-2 by Saccharomyces cerevisiae: use of a general secretion vector containing promoter and leader sequences of the mating pheromone α-factor. Gene 37: 155–161.

    Article  CAS  Google Scholar 

  9. Brake, A.J., Merryweather, J.P., Coit, D.G., Heberlein, U.A., Masiarz, F.R., Mullenbach, G.T., Urdea, M.S., Valenzuela, P. and Barr, P.J. 1984. α-factor-directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae. Proc. Nat. Acad. Sci. USA 81: 4642–4646.

    Article  CAS  Google Scholar 

  10. Miyajima, A., Otsu, K., Schreurs, J., Bond, M.W., Abrams, J.S. and Arai, K. 1986. Expression of murine and human granulocyte-macrophage colony-stimulating factors in S. cerevisiae: mutagenesis of the potential glycosylation sites. EMBO J. 5: 1193–1197.

    Article  CAS  Google Scholar 

  11. Kniskern, P.J., Hagopian, A., Montgomery, D.L., Burke, P., Dunn, N.R., Hofmann, K.J., Miller, W.J. and Ellis, R.W. 1986. Unusually high-level expression of a foreign gene (hepatitis B virus core antigen) in Saccharomyces cerevisiae. Gene 46: 135–141.

    Article  CAS  Google Scholar 

  12. Bennetzen, J.L. and Hall, B.D. 1981. Codon selection in yeast. J. Biol. Chem. 257: 3026–3031.

    Google Scholar 

  13. Ikemura, T. 1982. Correlation between the abundance of yeast transfer RNAs and the occurrence of the respective codons in protein genes. J. Mol. Biol. 158: 573–597.

    Article  CAS  Google Scholar 

  14. Hoekema, A., Kastelein, R.A., Vasser, M. and di Boer, H.A. 1987. Codon replacement in the PGK1 gene of Saccharomyces cerevisiae: Experimental approach to study the role of biased codon usage in gene expression. Mol. Cell Biol. 7: 2914–2924.

    Article  CAS  Google Scholar 

  15. Ngsee, J.K. and Smith, M. 1990. Changes in a mammalian signal sequence required for efficient protein secretion by yeasts. Gene 86: 251–255.

    Article  CAS  Google Scholar 

  16. Urdea, M.S., Merryweather, J.P., Mullenbach, G.T., Coit, D., Heberlein, U., Valenzuela, P. and Barr, P.J. 1983. Chemical synthesis of a gene for human epidermal growth factor urogastrone and its expression in yeast. Proc. Nat. Acad. Sci. USA 80: 7461–7465.

    Article  CAS  Google Scholar 

  17. Sun, L.K., Curtis, P., Rakowicz-Szulczynska, E., Ghrayeb, J., Chang, N., Morrison, S.L. and Koprowski, H. 1987. Chimeric antibody with human constant regions and mouse variable regions directed against carcinoma-associated antigen 17-1A. Proc. Nat. Acad. Sci. USA 84: 214–218.

    Article  CAS  Google Scholar 

  18. Julius, D., Schekman, R. and Thorner, J. 1984. Glycosylation and processing of prepro-α-factor through the yeast secretory pathway. Cell 36: 309–318.

    Article  CAS  Google Scholar 

  19. Elliott, S., Giffin, J., Suggs, S., Lau, E.P. and Banks, A.R. 1989. Secretion of glycosylated human erythropoietin from yeast directed by the α-factor leader region. Gene 79: 167–180.

    Article  CAS  Google Scholar 

  20. Egel-Mitani, M., Hansen, M.T., Norris, K., Snel, L. and Fiil, N.P. 1988. Competitive expression of two heterologous genes inserted into one plasmid in Saccharomyces cerevisiae. Gene 73: 113–120.

    Article  CAS  Google Scholar 

  21. Ecker, D.J., Butt, T.R., Sternberg, E.J., Neeper, M.P., Debouck, C., Gorman, J.A. and Crooke, S.T. 1986. Yeast metallothionein function in metal ion detoxification. J. Biol. Chem. 261: 16895–16900.

    CAS  PubMed  Google Scholar 

  22. Sherman, F., Fink, G.R. and Hicks, J.B. 1982. Methods in Yeast Genetics: A Laboratory Manual. Cold Spring Harbor Laboratory, NY.

    Google Scholar 

  23. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, NY.

    Google Scholar 

  24. Taussig, R. and Carlson, M. 1983. Nucleotide sequence of the yeast SUC2 gene for invertase. Nuc. Acids Res. 11: 1943–1954.

    Article  CAS  Google Scholar 

  25. Harlow, E. and Lane, D. 1988. Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, NY.

    Google Scholar 

  26. Chomczynski, P. and Sacchi, N. 1986. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162: 156–159.

    Article  Google Scholar 

  27. Zamb, T.J. and Petes, T.D. 1982. Analysis of the junction between ribosomal RNA genes and single-copy chromosomal sequences in the yeast Saccharomyces cerevisiae. Cell 28: 355–364.

    Article  CAS  Google Scholar 

  28. Butt, T.R., Sternberg, E.J., Gorman, J.A., Clark, P., Hamer, M., Rosenberg, M. and Crooke, S.T. 1984. Copper metallothionein of yeast, structure of the gene, and regulation of expression. Proc. Nat. Acad. Sci. USA 81: 3332–3336.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Peter J. Curtis: Corresponding author.

Authors and Affiliations

  1. The Wistar Institute of Anatomy and Biology, 3601 Spruce Street, Philadelphia, PA, 19104

    Leszek Kotula & Peter J. Curtis

Authors
  1. Leszek Kotula
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter J. Curtis
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kotula, L., Curtis, P. Evaluation of Foreign Gene Codon Optimization in Yeast: Expression of a Mouse IG Kappa Chain. Nat Biotechnol 9, 1386–1389 (1991). https://doi.org/10.1038/nbt1291-1386

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt1291-1386

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing