Abstract
The efficient production of proteins via expression of recombinant genes in cells other than those that naturally produce the protein has been critical to the study and the therapeutic utility of a wide variety of proteins. E. coli expression systems are useful for many proteins. However, large complex proteins are not expressed efficiently in bacteria. Also, bacterial expression cannot be used for eukaryotic proteins that require glycosylation for their activity. Some established animal cell lines can be used as production hosts since they are easily transfected, can grow to high cell densities, and can be propagated in low-serum or serum-free medium.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kaufman, R. J., and Sharp, P. A. (1982) Amplification and expression of sequences cotransfected with a modular dihydrofolate reductase complementary DNA gene. J. Mol. Biol. 159, 601–621.
Haynes, J., and Weissmann, C. (1983) Constitutive long-term production of human interferons by hamster cells containing multiple copies of a cloned interferon gene. Nucleic Acids Res. 11, 687–707.
Scahill, S. J., Devos, R., Van der Heyden, J., and Fiers, W. (1983) Expression and characterization of the product of a human immune interferon cDNA gene in Chinese hamster ovary cells. Proc. Natl. Acad. Sci. USA 80, 4654–4658.
McCormick, F., Trahey, M., Innis, M., Dieckmann, B., and Ringold, G. (1984) Inducible expression of amplified human beta interferon genes in CHO cells. Mol. Cell. Biol. 4, 166–172.
Kaufman, R. J., Wasley, L. C., Spiliotes, A. J., Gossels, S. D., Latt, S. A., Larsen, G. R., and Kay, R. M. (1985) Coamplification and expression of human tissue-type plasminogen activator and murine dihydrofolate reductase sequences in Chinese hamster ovary cells. Mol. Cell. Biol 5, 1750–1759.
Urlaub, G. and Chasin, L. A. (1980) Isolation of Chinese hamster cell mutants deficient in dihyrofolate reductase activity. Proc. Natl. Acad. Sci. USA 77, 4216–4220.
Barsoum, J. (1990) Introduction of stable high-copy-number DNA into Chinese hamster ovary cells by electroporation. DNA Cell Biol. 9, 293–300.
Sato, K., Ito, R., Baek, K.-H., and Agarwal, K. (1986) A specific DNA sequence controls termination of transcription in the gastrin gene. Mol. Cell Biol. 6, 1032–1043.
Fisher, R. A., et al. (1988) HIV infection is blocked in vitro by recombinant soluble CD4. Nature 331, 76–78.
Miller, G. T., et al. (1993) Specific interaction of lymphocyte function-associated antigen 3 with CD2 can inhibit T cell responses. J. Exp. Med. 178, 211–222.
Lobb, R., et al. (1991) Expression and functional characterization of a soluble form of vascular cell adhesion molecule 1. Biochem. Biophys. Res. Commun. 178, 1498–1504.
Graham, F. L., and Van der Eb, A. J. (1973) A new technique for the assay of infectivity of human adenovirus 5. Virology 52, 456–467.
Bebbington, C. R., Renner, G., Thomson, S., King, D., Abrams, D., and Yarranton, G. T. (1992) High-level expression of a recombinant antibody from myeloma cells using a glutamine synthetase gene as an amplifiable selectable marker. Bio/Technology 10, 169–175.
Southern, P. J. and Berg, P. (1982) Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region. J. Mol. Appl. Genet. 1, 327–341.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Humana Press Inc.
About this protocol
Cite this protocol
Barsoum, J. (1995). Stable Integration of Vectors at High Copy Number for High-Level Expression in Animal Cells. In: Nickoloff, J.A. (eds) Animal Cell Electroporation and Electrofusion Protocols. Methods in Molecular Biology, vol 48. Humana Press. https://doi.org/10.1385/0-89603-304-X:225
Download citation
DOI: https://doi.org/10.1385/0-89603-304-X:225
Publisher Name: Humana Press
Print ISBN: 978-0-89603-304-7
Online ISBN: 978-1-59259-535-8
eBook Packages: Springer Protocols