Abstract
The expression of cloned genes in prokaryotic or eukaryotic host cells provides the means not only for the study of gene function but also for the production of substantial amounts of protein and nonprotein molecules for commercial and investigational use. In the case of proteins, strategies for determining the most appropriate vector-host combination for the expression of an exogenous gene depend on a diverse range of factors that relate ultimately to the properties of the gene and its product. The approach used in the downstream purification of the product is another factor that impinges on this selection. However, among the most important considerations in the choice of vector and host in ensuring the maximal amount of expression is the compatibility of the host cells to translate the RNA transcript, to ensure the proper folding of the product, and to sustain the protein in the intact and functional state.
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
Baneyx, F. (1999) Recombinant protein expression in Escherichia coli. Curr. Opin. Biotechnol. 10, 411–421.
Balbás, P. (2001) Understanding the art of producing protein and nonprotein molecules in Escherichia coli. Mol. Biotechnol. 19, 251–267.
Swartz, J. R. (2001) Advances in Escherichia coli production of therapeutic proteins. Curr. Opin. Biotechnol. 12, 195–201.
Jonasson, P., Lijeqvist, S., Nygren, P., and Ståhl, S. (2002) Genetic design for facilitated production and recovery of recombinant proteins in Escherichia coli. Biotechnol. Appl. Biochem. 35, 91–105.
Kaufman, R. J. (2000) Overview of vector design for mammalian gene expression. Mol. Biotechnol. 16, 151–160.
de Boer, H. A. and Kastelein, R. A. (1986) Biased codon usage: an exploration of its role in optimization of translation, in Maximizing Gene Expression (Reznikoff, W. and Gold, L., eds.), Buttersworth, Boston.
Akashi, H. (2001) Gene expression and molecular evolution. Curr. Opin. Genet. Dev. 11, 660–666.
Kane J. F. (1995) Effects of rare codon clusters on high-level expression of heterologous proteins in Escherichia coli. Curr. Opin. Biotechnol. 6, 494–500.
Hannig, G. and Makrides, S. C. (1998) Strategies for optimizing heterologous protein expression in Escherichia coli. Trends in Biotechn. 16, 54–60.
Prinz, W. A., Aslund, F., Holmgren, A., and Beckwith, J. (1997) The role of the thioredoxin and glutaredoxin pathways in reducing protein disulfide bonds in the Escherichia coli cytoplasm. J. Biol. Chem. 272, 15661–15667.
LaVallie, E. R., DiBlasio-Smith, E. A., Collins-Racie, L. A., Lu, Z. and McCoy, J. M. (2003). Thioredoxin and related proteins as multifunctional fusion tags for soluble expression in E. coli. Methods Mol. Biol. 205, 119–140.
Lobel, L., Pollak, S., Klein, J., and Lustbader, J.W. (2001) High-level bacterial expression of a natively folded, soluble extracellular domain fusion protein of the human luteinizing hormone/chorionic gonadotropin receptor in the cytoplasm of Escherichia coli. Endocrine 14, 205–212.
Lobel, L., Pollak, S., Lustbader, B., Klein, J., and Lustbader, J.W. (2002) Bacterial expression of a natively folded extracellular domain fusion protein of the hFSH receptor in the cytoplasm of Escherichia coli. Protein Express. Purif. 25, 124–133.
Wilkinson, D. L. and Harrison, R. G. (1991) Predicting the solubility of recombinant proteins in Escherichia coli. Biotechnology 9, 443–448.
Davis, G. D., Elisee, C., Newham, D. M. and Harrison, R. G. (1999) New fusion protein systems designed to give soluble expression in Escherichia coli. Biotechnol Bioeng. 65, 382–388.
Romanos, M. A., Scorer, C. A. and Clare, J. J. (1992) Foreign gene expression in yeast: a review. Yeast 8, 423–488.
Sudbery, P. E. (1996) The expression of recombinant proteins in yeasts. Curr. Opin. Biotechnol. 7, 517–524.
Ostergaard, S., Olsson, L., and Nielsen, J. (2000) Metabolic engineering of Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 64, 34–50.
Giga-Hama, Y. and Kumagai, H. (1999) Expression system for foreign genes using the fission yeast Schizosaccharomyces pombe. Biotechnol. Appl. Biochem. 30, 235–244.
Luckow, V. A. (1993) Baculovirus systems for the expression of human gene products. Curr. Opin. Biotechnol. 4, 564–572.
Jarvis, D. L. and Guarino, L. A. (1995) Continuous foreign gene expression in transformed lepidopteran insect cells. Methods Mol. Biol. 39, 187–202.
McCarroll, L. and King, L. A. (1997) Stable insect cell cultures for recombinant protein production. Curr. Opin. Biotechnol. 8, 590–594.
Jarvis, D. L. and Finn, E. E. (1995) Biochemical analysis of the N-glycosylation pathway in baculovirus-infected lepidopteran insect cells. Virology 212, 500–511.
Jarvis, D. L., Howe, D. and Aumiller, J. J. (2001) Novel baculovirus expression vectors that provide sialylation of recombinant glycoproteins in lepidopteran insect cells. J. Virol. 75, 6223–6227.
Condreay, J. P., Witherspoon, S. M., Clay, W. C. and Kost, T. A. (1997) Transient and stable gene expression in mammalian cells transduced with a recombinant baculovirus vector. Proc. Natl. Acad. Sci. USA 96, 127–132.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Greene, J.J. (2004). Host Cell Compatibility in Protein Expression. In: Balbás, P., Lorence, A. (eds) Recombinant Gene Expression. Methods in Molecular Biology, vol 267. Humana Press. https://doi.org/10.1385/1-59259-774-2:003
Download citation
DOI: https://doi.org/10.1385/1-59259-774-2:003
Publisher Name: Humana Press
Print ISBN: 978-1-58829-262-9
Online ISBN: 978-1-59259-774-1
eBook Packages: Springer Protocols