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 axis from 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.
CITATION STYLE
Cho, M. S., Yee, H., Brown, C., Jeang, K. T., & Chan, S. (2001). An oriP expression vector containing the HIV-1 Tat/TAR transactivation axis produces high levels of protein expression in mammalian cells. Cytotechnology, 37(1), 23–30. https://doi.org/10.1023/A:1016124911071
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