COS Cell Expression

  • John F. Hancock
Part of the Methods in Molecular Biology book series (MIMB, volume 8)


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.


Transfection Efficiency Cesium Chloride Calcium Phosphate Precipitation Defective Mutant Shock Solution 
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5. References

  1. 1.
    Gluzman, Y. (1981) SV40 transformed simian cells support the replication of early SV40 mutants. Cell 23, 175–182.PubMedCrossRefGoogle Scholar
  2. 2.
    D’Andrea, A. D., Lodish, H. F., and Wong, G. G (1989) Expression cloning of the murine erythropoietin receptor. Cell 57, 277–285.PubMedCrossRefGoogle Scholar
  3. 3.
    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.CrossRefGoogle Scholar
  4. 4.
    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, 825–828PubMedCrossRefGoogle Scholar
  5. 5.
    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.PubMedGoogle Scholar
  6. 6.
    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.PubMedCrossRefGoogle Scholar
  7. 7.
    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.PubMedCrossRefGoogle Scholar
  8. 8.
    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.PubMedCrossRefGoogle Scholar
  9. 9.
    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.PubMedGoogle Scholar
  10. 10.
    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.PubMedCrossRefGoogle Scholar
  11. 11.
    Seed, B. (1987) An LFA-3 cDNA encodes a phosphohpid linked membrane protein homologous to its receptor CD2. Nature 329, 840–842.PubMedCrossRefGoogle Scholar
  12. 12.
    Maniatis, T., Fritisch, E. F., and Sambrook, J. (1982) Molecular Cloning. A Laboratory Manual (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY)Google Scholar

Copyright information

© The Humana Press Inc., Clifton, NJ 1991

Authors and Affiliations

  • John F. Hancock
    • 1
  1. 1.Department ofHaematologyRoyal Free Hospital School of Medicine, HampsteadLondonUK

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