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COS Cell Expression

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

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.

Keywords

Transfection Efficiency Cesium Chloride Calcium Phosphate Precipitation Defective Mutant Shock Solution 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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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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