Human erythropoietin gene: high level expression in stably transfected mammalian cells and chromosome localization.
Human erythropoietin gene: high level expression in stably transfected mammalian cells and chromosome localization.
复制标题
人促红细胞生成素基因：在稳定转染的哺乳动物细胞中高水平表达和染色体定位。
DOI：
10.1073/pnas.83.17.6465
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发表时间：
1986
影响因子：
11.1
通讯作者：
Adamson,JW
中科院分区：
文献类型：
--
作者：
Powell,JS;Berkner,KL;Lebo,RV;Adamson,JW
糖蛋白激素促红细胞生成素在调节红细胞生成中起主要作用，促红细胞生成素缺乏会导致贫血。由于缺乏纯化材料，对这种激素的详细研究和替代疗法的尝试一直很困难。我们利用克隆的人促红细胞生成素基因建立了稳定转基因的哺乳动物细胞系，该细胞系能分泌大量具有强大生物学活性的激素。这些细胞系是通过将含有可选择标记的质粒和含有克隆的人促红细胞生成素基因的质粒结构插入到强启动子旁边，与哺乳动物细胞共转染而产生的。这些细胞分泌的蛋白质刺激红系祖细胞的增殖和分化，随着选择的增加，其中几种细胞株每升上清液分泌高达80毫克的蛋白质。对高分辨率双激光分选分离的人染色体DNA的杂交分析表明，人促红细胞生成素基因位于人类7号染色体上。
The glycoprotein hormone erythropoietin plays a major role in regulating erythropoiesis and deficiencies of erythropoietin result in anemia. Detailed studies of the hormone and attempts at replacement therapy have been difficult due to the scarcity of purified material. We used a cloned human erythropoietin gene to develop stably transfected mammalian cell lines that secrete large amounts of the hormone with potent biological activity. These cell lines were produced by cotransfection of mammalian cells with a plasmid containing a selectable marker and plasmid constructions containing a cloned human erythropoietin gene inserted next to a strong promoter. The protein secreted by these cells stimulated the proliferation and differentiation of erythroid progenitor cells and, with increased selection, several of these cell lines secrete up to 80 mg of the protein per liter of supernatant. Hybridization analysis of DNA from human chromosomes isolated by high resolution dual laser sorting provides evidence that the gene for human erythropoietin is located on human chromosome 7.