Yoshimoto T, Yamamoto Y, Arakawa T, Suzuki H, Yamamoto S, Yokoyama C, Tanabe T, Toh H
Department of Biochemistry, Tokushima University School of Medicine, Japan.
Biochem Biophys Res Commun. 1990 Nov 15;172(3):1230-5. doi: 10.1016/0006-291x(90)91580-l.
The cDNA for a 12-lipoxygenase was isolated from cDNA library of human erythroleukemia cells. The cDNA had an open reading frame encoding 663 amino acids with a calculated molecular weight of 75,513. The deduced amino acid sequence of human 12-lipoxygenase exhibited 41.5%, 65.3% and 65.4% identity with human 5-lipoxygenase, human 15-lipoxygenase and porcine 12-lipoxygenase, respectively. Blot hybridization analysis of RNA from human erythroleukemia cells demonstrated a single species (3.1 kb) of mRNA with the cDNA probe for 12-lipoxygenase of these cells, but not with the cDNA for porcine leukocyte enzyme. The cytosol of Escherichia coli transformed with a recombinant pUC19 plasmid oxygenated the position 12 of arachidonic acid.
从人红白血病细胞的cDNA文库中分离出12-脂氧合酶的cDNA。该cDNA有一个开放阅读框，编码663个氨基酸，计算分子量为75513。推导的人12-脂氧合酶氨基酸序列与人类5-脂氧合酶、人类15-脂氧合酶和猪12-脂氧合酶的一致性分别为41.5%、65.3%和65.4%。用人红白血病细胞的RNA进行印迹杂交分析表明，用这些细胞的12-脂氧合酶cDNA探针可检测到一种单一的mRNA（3.1kb），而用猪白细胞酶的cDNA则检测不到。用重组pUC19质粒转化的大肠杆菌细胞质可使花生四烯酸的12位发生氧化。
