Research Paper | Published:

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

Bio/Technology volume 9, pages 13861389 (1991) | Download Citation

Subjects

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    and 1988. Assembly of a functional immunoglobulin Fv fragment in Escherichia coli. Science 240: 1038–1041.

  2. 2.

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

  3. 3.

    and 1989. Expression of engineered antibodies and antibody fragments in microorganisms. Methods in Enzymol. 178: 476–496.

  4. 4.

    and 1990. High-level production of a functional immunoglobulin heterodimer in a baculovirus expression system. Proc. Nat. Acad. Sci. USA 87: 3942–3946.

  5. 5.

    , , , , and 1990. Antibody production in baculovirus-infected insect cells. Bio/Technology 8: 651–654.

  6. 6.

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

  7. 7.

    , , , , , , , and 1986. Protein secretion from Saccharomyces cerevisiae directed by the prepro-α-factor leader region. J. Biol. Chem. 261: 5858–5865.

  8. 8.

    , , , and 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.

  9. 9.

    , , , , , , , and 1984. α-factor-directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae. Proc. Nat. Acad. Sci. USA 81: 4642–4646.

  10. 10.

    , , , , and 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.

  11. 11.

    , , , , , , and 1986. Unusually high-level expression of a foreign gene (hepatitis B virus core antigen) in Saccharomyces cerevisiae. Gene 46: 135–141.

  12. 12.

    and 1981. Codon selection in yeast. J. Biol. Chem. 257: 3026–3031.

  13. 13.

    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.

  14. 14.

    , , and 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.

  15. 15.

    and 1990. Changes in a mammalian signal sequence required for efficient protein secretion by yeasts. Gene 86: 251–255.

  16. 16.

    , , , , , and 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.

  17. 17.

    , , , , , and 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.

  18. 18.

    , and 1984. Glycosylation and processing of prepro-α-factor through the yeast secretory pathway. Cell 36: 309–318.

  19. 19.

    , , , and 1989. Secretion of glycosylated human erythropoietin from yeast directed by the α-factor leader region. Gene 79: 167–180.

  20. 20.

    , , , and 1988. Competitive expression of two heterologous genes inserted into one plasmid in Saccharomyces cerevisiae. Gene 73: 113–120.

  21. 21.

    , , , , , and 1986. Yeast metallothionein function in metal ion detoxification. J. Biol. Chem. 261: 16895–16900.

  22. 22.

    , and 1982. Methods in Yeast Genetics: A Laboratory Manual. Cold Spring Harbor Laboratory, NY.

  23. 23.

    , and 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, NY.

  24. 24.

    and 1983. Nucleotide sequence of the yeast SUC2 gene for invertase. Nuc. Acids Res. 11: 1943–1954.

  25. 25.

    and 1988. Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, NY.

  26. 26.

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

  27. 27.

    and 1982. Analysis of the junction between ribosomal RNA genes and single-copy chromosomal sequences in the yeast Saccharomyces cerevisiae. Cell 28: 355–364.

  28. 28.

    , , , , , and 1984. Copper metallothionein of yeast, structure of the gene, and regulation of expression. Proc. Nat. Acad. Sci. USA 81: 3332–3336.

Download references

Author information

Author notes

    • Peter J. Curtis

    Corresponding author.

Affiliations

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

    • Leszek Kotula
    •  & Peter J. Curtis

Authors

  1. Search for Leszek Kotula in:

  2. Search for Peter J. Curtis in:

About this article

Publication history

Received

Accepted

Published

DOI

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

Further reading