Miller D B, Munster D, Wasvary J S, Simke J P, Peppard J V, Bowen B R, Marshall P J
Department of Biological Technologies, Ciba Pharmaceuticals Division, CIBA-GEIGY Corporation, Summit, NJ 07901.
Biochem Biophys Res Commun. 1994 May 30;201(1):356-62. doi: 10.1006/bbrc.1994.1709.
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.
通过对白细胞介素-1β刺激的人皮肤成纤维细胞cDNA进行PCR扩增，克隆出人类环氧化酶-2的开放阅读框。编码区用于构建重组杆状病毒，该病毒用于感染Sf9细胞时可指导重组人环氧化酶-2的表达。对异源表达的酶进行了表征，发现其具有环氧化酶的所有显著特征。感染细胞的大规模微粒体制备物产生了超过20个酶单位，比活性为240纳摩尔前列腺素产物/毫克蛋白质。
