Unit

UNIT 16.23 Amplification Using CHO Cell Expression Vectors

  1. Robert E. Kingston1,
  2. Randal J. Kaufman (DHFR)2,
  3. C.R. Bebbington3,
  4. M.R. Rolfe (GS)3

Published Online: 1 NOV 2002

DOI: 10.1002/0471142727.mb1623s60

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Kingston, R. E., Kaufman, R. J., Bebbington, C. and Rolfe, M. 2002. Amplification Using CHO Cell Expression Vectors. Current Protocols in Molecular Biology. 60:III:16.23:16.23.1–16.23.13.

Author Information

  1. 1

    Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts

  2. 2

    University of Michigan, Ann Arbor, Michigan

  3. 3

    Celltech Ltd., Slough, United Kingdom

Publication History

  1. Published Online: 1 NOV 2002
  2. Published Print: OCT 2002

Abstract

The ability to select for integration of plasmid DNA into the host chromosome allows the generation of stably transfected cell lines. With transfection of a selectable marker linked to a nonselectable target gene (or by cotransfection of the two unlinked genes), high-level expression of the desired gene is obtained by selecting for amplification of the selectable marker. This unit presents two systems for gene amplification and expression. The first describes the dihydrofolate reductase (DHFR) selection system while the second is based on selection of the glutamine synthetase (GS) gene. The DHFR system is probably more widely used, and results in very high levels of amplification and expression; however, the DHFR amplification process is lengthy and may require several months to isolate and characterize a stable, amplified line. In contrast, the GS system typically requires only a single round of selection for amplification to achieve maximal expression levels.