The heterologous expression and characterization of human prostaglandin G/H synthase-2 (COX-2)

Biochem Biophys Res Commun. 1994 May 30;201(1):356-62. doi: 10.1006/bbrc.1994.1709.

Abstract

The open reading frame of human cyclooxygenase-2 was cloned by pcr amplification of IL-1 beta stimulated human dermal fibroblast cDNA. The coding region was used to construct a recombinant baculovirus which when used to infect Sf9 cells directed the expression of recombinant human cyclooxygenase-2. The heterologously expressed enzyme was characterized and found to display all salient features of cyclooxygenase. Large-scale microsomal preparations of infected cells yielded more than 20 units of enzyme with a specific activity of 240 nmoles prostaglandin product/mg protein.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Compartmentation
  • Cloning, Molecular
  • Cyclooxygenase Inhibitors / pharmacology
  • DNA Primers / chemistry
  • Gene Expression
  • Humans
  • Molecular Sequence Data
  • Moths
  • Prostaglandin-Endoperoxide Synthases / genetics*
  • Prostaglandin-Endoperoxide Synthases / metabolism
  • Prostaglandins / biosynthesis
  • RNA, Messenger / genetics
  • Recombinant Proteins

Substances

  • Cyclooxygenase Inhibitors
  • DNA Primers
  • Prostaglandins
  • RNA, Messenger
  • Recombinant Proteins
  • Prostaglandin-Endoperoxide Synthases