The heterologous expression and characterization of human prostaglandin G/H synthase-2 (COX-2).
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.
Miller
D B
DB
Department of Biological Technologies, Ciba Pharmaceuticals Division, CIBA-GEIGY Corporation, Summit, NJ 07901.
Munster
D
D
Wasvary
J S
JS
Simke
J P
JP
Peppard
J V
JV
Bowen
B R
BR
Marshall
P J
PJ
D000818:Animals; D001483:Base Sequence; D002451:Cell Compartmentation; D003001:Cloning, Molecular; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D009036:Moths; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics* / Q000378:metabolism; D011453:Prostaglandins / Q000096:biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins
D016861:Cyclooxygenase Inhibitors; D017931:DNA Primers; D011453:Prostaglandins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D011451:Prostaglandin-Endoperoxide Synthases