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作者通过RT-PCR和引物延伸PCR得到了全长的人类环加氧酶-2(hcox-2)的cDNA,所推测的hcox-2多肽氨基酸顺序与hcox-1蛋白的氨基酸顺序有61％同源,而与鸡、鼠cox-2基因所编码的多肽分别有81％和88％同源。作者又将hcox-1、hcox-2cDNA的 DRFs亚克隆在真核表达载体pcDNA中,所得到的相应mRNA在~(35)S-甲硫氨酸兔网织红细胞裂解物中进行翻译,合成了70kD的多肽。然后分别把hcox-1和hcox-2 cDNA转染到COS细胞中,它们都表达了环加氧酶活性。这些资料表明hcox-2cDNA能够产生有酶活性的环加氧酶。作者进一步
The full-length cDNA of human cyclooxygenase-2 (hcox-2) was obtained by RT-PCR and primer extension PCR. The predicted amino acid sequence of hcox-2 polypeptide was 61% identical to the amino acid sequence of hcox-1 protein Source, and with chicken, mouse cox-2 gene encoded polypeptide 81% and 88% homologous, respectively. The authors further subcloned hcox-1, hcox-2 cDNA DRFs in the eukaryotic expression vector pcDNA, the corresponding mRNA obtained in ~ (35) S-methionine rabbit reticulocyte lysate translation, synthesis 70kD polypeptide. The hcox-1 and hcox-2 cDNAs were then transfected into COS cells, respectively, which all expressed cyclooxygenase activity. These data indicate that hcox-2 cDNA is capable of producing enzymatically active cyclooxygenase. Author further