Abstract
COS-1 cells were created by transforming an established line of monkey epithelial cells, CV-1, with a defective mutant of SV40 (1). The SV40 mutant used carried a small deletion within the origin of replication and, although this construct transformed CV-1 cells, which are permissive for lytic growth of SV40, no infectious virus was produced after prolonged culture. One transformed cell line, COS-1, was fully characterized and found to contain the complete early region of the SV40 genome. COS-1 cells express nuclear large T and all proteins necessary for replication of appropriate circular genomes. This was first demonstrated by showing that COS-1 cells could support the replication of early region mutants of SV40. More important, however, the introduction of any plasmid containing an SV40 origin of replication into COS-1 cells results in rapid replication of the plasmid to high copy number. Coincidently, of course, the transfected cells will express any gene on the plasmid that is driven by a suitable eukaryotic promoter. The combined effect of these phenomena is transient high-level expression of the encoded protein.
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
5. References
Gluzman, Y. (1981) SV40 transformed simian cells support the replication of early SV40 mutants. Cell 23, 175–182.
D’Andrea, A. D., Lodish, H. F., and Wong, G. G (1989) Expression cloning of the murine erythropoietin receptor. Cell 57, 277–285.
Aruffo. A. and Seed. B. (1987) Molecular cloning of aCD28cDNAby a high effficiency COS cell expression system. Proc. Natl. Acad. Sci. USA 84, 8575–8577.
Yamasaki, X., Taga, T., Hirata, Y., Yawata, H., Kawanishi, Y., Seed, B., Taniguchi, T., Hirano, T., and Kishimoto, T. (1988) Cloning and expression of the Human Interleukin-6 (BSF-2/IFN-2) Receptor Science 241, 825–828
Jing, S. Q. and Trowbridge, I. S. (1987) Identification of the lntermolecular disulphide bonds of the human transferrin receptor and its lipid attachment site. EMBO J. 6, 327–331.
Hancock, J. F., Magee, A. I, Childs, J., and Marshall, C. J. (1989) All ras proteins are polyisoprenylated but only some are palmitoylated. Cell 57, 1167–1177.
Reeves, R., Gorman, C, and Howard, B. (1985) Minichromosome aasembly of non-integrated plasmid DNA transfected into mammalian cells. Nucleic Acids Res 13, 3599–3615.
Lopata, M. A., Cleveland, D. W., and Sollner-Webb, B. (1984) High level expression of a chloramphenicol acetyltransferase gene by DEAE-dextran mediated DNA transfection coupled with a dimethyl sulphoxide or glycerol shock treatment. Nucleic Acids Res. 12, 5707–5717.
Hancock, J. F., Marshall, C. J., McKay, A. I., Gardner, S., Houslay, M. D, Hall, A., and Wakelam, M. J. O. (1988) Mutant but not normal p21™ elevates inositol phosphate breakdown in two different cell systems. Oncogene 3, 187–193.
Miller, J. and Germain, R.N. (1986) Efficientcell surface expression of class IIMHC molecules in the absence of associated invariant chain. J. Exp. Med. 164, 1478–1489.
Seed, B. (1987) An LFA-3 cDNA encodes a phosphohpid linked membrane protein homologous to its receptor CD2. Nature 329, 840–842.
Maniatis, T., Fritisch, E. F., and Sambrook, J. (1982) Molecular Cloning. A Laboratory Manual (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY)
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1991 The Humana Press Inc., Clifton, NJ
About this protocol
Cite this protocol
Hancock, J.F. (1991). COS Cell Expression. In: Collins, M.K.L. (eds) Practical Molecular Virology. Methods in Molecular Biology, vol 8. Humana Press. https://doi.org/10.1385/0-89603-191-8:153
Download citation
DOI: https://doi.org/10.1385/0-89603-191-8:153
Publisher Name: Humana Press
Print ISBN: 978-0-89603-191-3
Online ISBN: 978-1-59259-495-5
eBook Packages: Springer Protocols