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[人促红细胞生成素受体胞外区cDNA的克隆及其在大肠杆菌中的表达]

[Cloning cDNA of extracellular domain of human erythropoietin receptor and its expression in Escherichia coli].

作者信息

Zhang Y H, Lu Y F, Liu Y P, Deng J X

机构信息

Beijing Institute of Biotechnology.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2000 May;16(3):328-32.

Abstract

Human erythropoietin receptor (hEpoR) plays an important role in regulating the red blood cell production by promoting the proliferation and differentiation of RBC from erythroid precursors. hEpoR is a transmembrane protein, and its extrocellular domain (sEpoR) is of great importance in Epo signal transduction pathway. We cloned the gene of sEpoR by RT-PCR from the total RNA of human fetal liver and expressed it in E. coli after insertion of the gene in the expression vector pBV220. The cloned gene was confirmed by sequencing analysis and gene product was confirmed by both Western blot and its first 11 amino acid residues sequence of the N-terminal. In vitro bioassay showed that the purified gene product can repress the growth of TF-cells in the presence of Epo.

摘要

人促红细胞生成素受体(hEpoR)通过促进红细胞前体中红细胞的增殖和分化,在调节红细胞生成中发挥重要作用。hEpoR是一种跨膜蛋白,其胞外结构域(sEpoR)在促红细胞生成素(Epo)信号转导途径中至关重要。我们通过逆转录聚合酶链反应(RT-PCR)从人胎儿肝脏的总RNA中克隆了sEpoR基因,并将该基因插入表达载体pBV220后在大肠杆菌中进行表达。通过测序分析确认了克隆的基因,并通过蛋白质免疫印迹法(Western blot)及其N端的前11个氨基酸残基序列确认了基因产物。体外生物测定表明,纯化的基因产物在Epo存在的情况下可抑制TF细胞的生长。

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