THE HETEROLOGOUS EXPRESSION AND CHARACTERIZATION OF HUMAN PROSTAGLANDIN G/H SYNTHASE-2 (COX-2)

Abstract The open reading frame of human cyclooxygenase-2 was cloned by pcr amplification of IL-1β 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.