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Estrogen receptor beta yield from baculovirus lytic infection is higher than from stably transformed Sf21 cells

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Abstract

The production of estrogen receptors (ER) in cultured insect cells is advantageous because these cells are relatively easy to culture and they perform post-translation modifications necessary for protein stability and function. There are three options for protein expression in insect cells: transient transfection, lytic baculovirus infection, or transfection followed by selection to create stable cell lines. Stable transfection has been promoted to be advantageous for the production of recombinant proteins because no re-infection is required, which might provide better lot-to-lot reproducibility in protein production. In this paper, we demonstrate that lytic baculovirus infection of Sf21 cells yields approximately tenfold more bioactive ERβ than cells stably transformed with pIZ/V5-His plasmid under OpIE2 promoter. We provide the first evidence that stable expression of recombinant human ERβ decreases the proliferation of Sf21 cells by inhibition of cell replication in a ligand-independent manner. These results mirror findings in breast cancer cells showing that an increase in ERβ expression decreases cell proliferation. We conclude that baculovirus infection of Sf21 cells is better for human ERβ production than stable-transformation of Sf21 cells.

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Acknowledgment

This research was supported by NIH R01 DK 53220 to CMK. We thank Dr. Samuel R. Wellhausen, Department of Pathology James Graham Brown Cancer Center, for his help with flow-cytometric analysis and Dr. Peter C. Kulakosky for his suggestions on this manuscript.

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  1. Department of Biochemistry & Molecular Biology and the Center for Genetics and Molecular Medicine, University of Louisville School of Medicine, Louisville, KY, 40292, USA

    Margarita M. Ivanova, Kathleen A. Mattingly & Carolyn M. Klinge

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  1. Margarita M. Ivanova
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  2. Kathleen A. Mattingly
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  3. Carolyn M. Klinge
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Correspondence to Carolyn M. Klinge.

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Ivanova, M.M., Mattingly, K.A. & Klinge, C.M. Estrogen receptor beta yield from baculovirus lytic infection is higher than from stably transformed Sf21 cells. Appl Microbiol Biotechnol 74, 1256–1263 (2007). https://doi.org/10.1007/s00253-006-0784-9

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  • DOI: https://doi.org/10.1007/s00253-006-0784-9

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