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Janmohamed A, Thaunsukon P, Shephard EA, et al. Expression of Recombinant Flavin-Containing Monooxygenases in a Baculovirus/Insect Cell System. Methods Mol Biol. 2006;320:39-59doi: 10.1385/1-59259-998-2:39.
Janmohamed, A., Thaunsukon, P., Shephard, E. A., & Phillips, I. R. (2006). Expression of Recombinant Flavin-Containing Monooxygenases in a Baculovirus/Insect Cell System. Methods in molecular biology (Clifton, N.J.), 32039-59. https://doi.org/10.1385/1-59259-998-2:39
Janmohamed, Azara, et al. "Expression of Recombinant Flavin-Containing Monooxygenases in a Baculovirus/Insect Cell System." Methods in molecular biology (Clifton, N.J.) vol. 320 (2006): 39-59. doi: https://doi.org/10.1385/1-59259-998-2:39
Janmohamed A, Thaunsukon P, Shephard EA, Phillips IR. Expression of Recombinant Flavin-Containing Monooxygenases in a Baculovirus/Insect Cell System. Methods Mol Biol. 2006;320:39-59. doi: 10.1385/1-59259-998-2:39. PMID: 27699678.
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Methods Mol Biol. 2006;320:39-59. doi: 10.1385/1-59259-998-2:39.

Expression of Recombinant Flavin-Containing Monooxygenases in a Baculovirus/Insect Cell System.

Methods in molecular biology (Clifton, N.J.)

Azara Janmohamed, Prapimpuk Thaunsukon, Elizabeth A Shephard, Ian R Phillips

Affiliations

  1. Department of Biochemistry and Molecular Biology, University College London, London, UK.
  2. School of Biological Sciences, Queen Mary, University of London, London, UK.

PMID: 27699678 DOI: 10.1385/1-59259-998-2:39

Abstract

The baculovirus/insect cell heterologous expression system provides an important tool for investigating the catalytic activity of individual drug-metabolizing enzymes toward a particular substrate. In this chapter we describe a baculovirus/insect cell system that we have used for the expression of human and mouse flavin-containing monooxygenases. Methods are described for the generation of recombinant baculoviral DNAs, via both site-specific transposition in Escherichia coli and site-specific recombination in vitro; adaptation of Spodoptera frugiperda (Sf) 9 cells to shaking culture and to serum-free medium; cryopreservation and transfection of Sf9 cells; amplification of baculovirus and determination of viral titer; analysis of baculoviral DNA; and expression and analysis of recombinant proteins.

Keywords: FMO4; Flavin-containing monooxygenase; baculovirus; cytochrome P450; heterologous expression; insect cells

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