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Cytotechnology

, Volume 37, Issue 1, pp 23–30 | Cite as

An oriP expression vector containing the HIV-1 Tat/TAR transactivation axis produces high levels of protein expression in mammalian cells

  • Myung-Sam ChoEmail author
  • Helena Yee
  • Colleen Brown
  • Kuan-Teh Jeang
  • Sam Chan
Article

Abstract

A mammalian gene expression vector based on cytomegalovirus (CMV)enhancer/promoter (CMVe/p) for the regulation of gene expression was further optimized by adding oriP elements derived from Epstein-Barr virus (EBV) and the Tat/TAR transactivation axisfrom human immunodeficiency virus type 1 (HIV-1). Using the Tat/TAR-oriP expression vector, a transient transfection system was optimized for an extended culture period to produce large amounts of secreted IL-2SA (an IL-2 mutein) in HKB11 cells. We observed a 4-fold increase in IL-2SA expression in cells transfected with vectors containing the HIV-1 transactivation axis (Tat/TAR) or oriP elements alone when compared to cells transfected with the control vector having a CMVe/p. Cells transfected with expression vectors equipped with both oriP and Tat/TAR showed an 18-fold increase in IL-2SA expression. This transient transfection system maintained high secretion of IL-2SA for a period of 10-day with no appreciable loss in expression. We demonstrate that during this 10-day culture period, it was possible to produce 1–100 mg of proteins using 500 μg of plasmid DNA.

HKB11 cells IL-2 mutein mammalian cellexpression oriTat/TAR 

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Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Myung-Sam Cho
    • 1
    Email author
  • Helena Yee
    • 2
  • Colleen Brown
    • 2
  • Kuan-Teh Jeang
    • 3
  • Sam Chan
    • 2
  1. 1.Molecular and Cell Biology, Process Sciences, BiotechnologyBayer CorporationBerkeleyU.S.A.
  2. 2.Molecular and Cell Biology, Process Sciences, BiotechnologyBayer CorporationBerkeleyU.S.A.
  3. 3.Molecular Virology Section, Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious DiseasesNational Institute of HealthBethesdaU.S.A.

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