Cloning and Expression of Cyclooxygenase-1 and Cyclooxygenase-2

  • Nicholas R. Staten
  • Beverly A. Reitz
Part of the Methods in Molecular Biology book series (MIMB, volume 644)


Cyclooxygenase (COX) enzymes play important roles in normal physiology and during inflammation of various cells and tissues. In order to help understand the functions of these enzymes, their genes can be cloned to facilitate the production of the proteins in recombinant form. We outline a method to clone the genes from a human macrophage cell line for expression in an insect cell line infected with recombinant baculovirus encoding these enzymes.

Key words

Cyclooxygenase (COX) Messenger ribonucleic acid (mRNA) Polymerase chain reaction (PCR) Baculovirus Bacmid Insect cells 


  1. 1.
    Shimokawa T, Smith WL (1992) Expression of prostaglandin endoperoxide synthase-1 in a baculovirus system. Biochem Biophys Res Commun 183(3):975–982CrossRefPubMedGoogle Scholar
  2. 2.
    Miller DB, Munster D, Wasvary JS, Simke JP, Peppard JV, Bowen BR, Marshall PJ (1994) The heterologous expression and characterization of human prostaglandin G/H synthase-2 (COX-2). Biochem Biophys Res Commun 201(1):356–362CrossRefPubMedGoogle Scholar
  3. 3.
    Barnett J, Chow J, Ives D, Chiou M, Mackenzie R, Osen E, Nguyen B, Tsing S, Bach C, Freire J, Chan J, Sigal E, Ramesha C (1994) Purification, characterization and selective inhibition of human prostaglandin G/H synthase 1 and 2 expressed in the baculovirus system. Biochim Biophys Acta 1209(1):130–139PubMedGoogle Scholar
  4. 4.
    Gierse JK, Hauser SD, Creely DP, Koboldt C, Rangwala SH, Isakson PC, Seibert K (1995) Expression and selective inhibition of the constitutive and inducible forms of human cyclo-oxygenase. Biochem J 305(Pt. 2):479–484PubMedGoogle Scholar
  5. 5.
    George HJ, Van Dyk DE, Straney RA, Trzaskos JM, Copeland RA (1996) Expression purification and characterization of recombinant human inducible prostaglandin G/H synthase from baculovirus-infected insect cells. Protein Expr Purif 7(1):19–26CrossRefPubMedGoogle Scholar
  6. 6.
    Gierse JK, Staten NR, Casperson GF, Koboldt CM, Trigg JS, Reitz BA, Pierce JL, Seibert K (2002) Cloning, expression, and selective inhibition of canine cyclooxygenase-1 and cyclooxygenase-2. Vet Ther 3(3):270–280PubMedGoogle Scholar
  7. 7.
    Zhang WY, Yang XN, Jin DZ, Zhu XZ (2004) Expression and enzyme activity determination of human cyclooxygenase-1 and -2 in a baculovirus-insect cell system. Acta Pharmacol Sin 25(8):1000–1006PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Nicholas R. Staten
    • 1
  • Beverly A. Reitz
    • 1
  1. 1.Pfizer, Inc.ChesterfieldUSA

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