Thiele B J, Fleming J, Chester J, O'Prey J, Prehn S, Janetzki S, Rapoport S M, Harrison P R
Institut für Biochemie, Humboldt-Universität, Berlin, DDR.
Biomed Biochim Acta. 1990;49(2-3):S17-24.
The complete structure of the rabbit erythroid cell-specific 15-lipoxygenase mRNA and its gene was established by sequencing cDNA and genomic recombinants. The transcription initiation site was obtained by primer-extension sequencing. A presumptive promoter structure was characterized by sequencing 0.5 kb 5' to the transcription initiation site and by transfection experiments using constructs with the chloramphenicol transferase gene. The mRNA codes for a polypeptide of 662 amino acids. Its 3' untranslated region contains an intriguing repeated sequence of 10 copies with the consensus C4PuC3TCTTC4AAG which may be involved in its regulation during reticulocyte maturation. The transcription unit consists of 8.0 kb and is split like the gene for the leukocyte 5' lipoxygenase by 13 introns. Another interesting aspect in the structure of the 15-LOX gene is a highly conserved repeat in intron seven consisting of a unit of 54 nucleotides which is repeated eight times. Comparing the predicted amino acid sequence with those from other lipoxygenases published recently shows that lipoxygenases are a related group of enzymes which may have arisen from a common ancestral gene.
通过对cDNA和基因组重组体进行测序,确定了兔红细胞特异性15 - 脂氧合酶mRNA及其基因的完整结构。转录起始位点通过引物延伸测序获得。通过对转录起始位点上游0.5 kb区域进行测序以及使用氯霉素转移酶基因构建体进行转染实验,对推定的启动子结构进行了表征。该mRNA编码一个662个氨基酸的多肽。其3'非翻译区包含一个由10个拷贝组成的有趣重复序列,共有序列为C4PuC3TCTTC4AAG,这可能在网织红细胞成熟过程中参与其调节。转录单元由8.0 kb组成,像白细胞5'脂氧合酶基因一样被13个内含子分割。15 - LOX基因结构的另一个有趣方面是内含子7中一个高度保守的重复序列,由一个54个核苷酸的单元重复八次组成。将预测的氨基酸序列与最近发表的其他脂氧合酶的序列进行比较表明,脂氧合酶是一组相关的酶,可能起源于一个共同的祖先基因。