CHARACTERIZATION OF HUMAN 12-LIPOXYGENASE GENES
CHARACTERIZATION OF HUMAN 12-LIPOXYGENASE GENES
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DOI：
10.1073/pnas.89.9.3962
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发表时间：
1992-05-01
影响因子：
11.1
通讯作者：
SAMUELSSON, B
中科院分区：
文献类型：
--
作者：
FUNK, CD;FUNK, LB;SAMUELSSON, B
从分离自三个基因组噬菌体和粘粒文库的13个不同克隆中鉴定了两个人12-脂氧合酶（花生四烯酸：氧12-氧化还原酶，EC 1.13.11.31）相关基因。一个完整的基因（12-脂氧合酶基因1）跨越几乎等于17个酶和14个外显子组成的序列匹配克隆的血小板/人红白血病（HEL）细胞的cDNA序列进行了鉴定。确定了转录因子的几个共有位点和5'侧翼区域内的两个潜在的转录起始位点，包括推定的启动子区域。第二，可能的假基因（12-脂氧合酶基因2），其中显示几乎等于85%的同一性基因1的外显子序列内的一段，也进行了表征。两个12-脂氧合酶基因的存在也证实了总人类基因组DNA的Southern印迹分析。12-脂氧合酶基因的外显子-内含子边界位于与先前克隆的人5-脂氧合酶和兔15-脂氧合酶基因相同的相应位置，表明高度相关的基因家族。三个脂氧合酶基因（12-脂氧合酶基因1和2，15-脂氧合酶）定位于人类17号染色体，而最无关的脂氧合酶（5-脂氧合酶）被定位于染色体10通过PCR分析的人-仓鼠体细胞杂交DNA面板。12-通过逆转录-PCR分析，可以在人红白血病细胞、血小板和人脐静脉内皮细胞中确定地检测到脂氧合酶基因1的表达。在几种组织和细胞系中没有检测到12-脂氧合酶基因2的表达。
Two human 12-lipoxygenase enzyme (arachidonate:oxygen 12-oxidoreductase, EC 1.13.11.31)-related genes were characterized from 13 distinct clones isolated from three genomic bacteriophage and cosmid libraries. A complete gene (12-lipoxygenase gene 1) spanning almost-equal-to 17 kilobases and consisting of 14 exons with sequence matching the cloned platelet/human erythroleukemia (HEL) cell cDNA sequence was identified. Several consensus sites for transcription factors and two potential transcription initiation sites within the 5' flanking region, encompassing the putative promoter region, were identified. A segment of a second, probable pseudogene (12-lipoxygenase gene 2), which displays almost-equal-to 85% identity to gene 1 within exon sequences, was also characterized. The presence of two 12-lipoxygenase genes was also substantiated by Southern blot analysis of total human genomic DNA. Exon-intron boundaries for the 12-lipoxygenase genes were located in the identical corresponding positions to the previously cloned human 5-lipoxygenase and rabbit 15-lipoxygenase genes, indicating a highly related gene family. Three lipoxygenase genes (12-lipoxygenase genes 1 and 2, 15-lipoxygenase) were localized to human chromosome 17, whereas the most unrelated lipoxygenase (5-lipoxygenase) was mapped to chromosome 10 by PCR analysis of a human-hamster somatic hybrid DNA panel. 12-Lipoxygenase gene 1 expression could be detected in human erythroleukemia cells, platelets, and human umbilical vein endothelial cells with certainty by reverse transcription-PCR analysis. There was no detectable 12-lipoxygenase gene 2 expression in several tissues and cell lines.