Expression cloning of the murine interferon gamma receptor cDNA.
Expression cloning of the murine interferon gamma receptor cDNA.
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DOI：
10.1073/pnas.86.23.9248
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发表时间：
1989-12
影响因子：
11.1
通讯作者：
Sean Munro;Tom Maniatis
中科院分区：
文献类型：
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作者：
Sean Munro;Tom Maniatis
从小鼠胸腺细胞制成的表达文库中分离出编码小鼠干扰素 γ (IFN-γ) 受体的 cDNA。通过将文库转染到猴 COS 细胞中并用放射性标记的鼠 IFN-γ 探测转染的单层来鉴定克隆。通过放射自显影术鉴定表达受体的细胞，并直接从产生阳性信号的那些细胞中拯救编码受体的质粒。使用如此获得的部分cDNA通过常规方法从小鼠L929细胞分离全长cDNA。当该 cDNA 在 COS 细胞中表达时，它会产生鼠 IFN-γ 的特异性结合位点，其亲和常数与 L929 细胞上发现的受体相似。鼠类 IFN-γ 受体的预测氨基酸序列与先前报道的人 IFN-γ 受体的氨基酸序列具有同源性。然而，尽管这两种蛋白明显相关，但它们在推定的胞外结构域和胞内结构域中的同一性均低于 60%。
A cDNA encoding a receptor for murine interferon gamma (IFN-gamma) was isolated from an expression library made from murine thymocytes. The clone was identified by transfecting the library into monkey COS cells and probing the transfected monolayer with radiolabeled murine IFN-gamma. Cells expressing the receptor were identified by autoradiography and plasmids encoding the receptor were directly rescued from those cells producing a positive signal. A partial cDNA so obtained was used to isolate a full-length cDNA from mouse L929 cells by conventional means. When this cDNA was expressed in COS cells it produced a specific binding site for murine IFN-gamma with an affinity constant similar to that of the receptor found on L929 cells. The predicted amino acid sequence of the murine IFN-gamma receptor shows homology to that previously reported for the human IFN-gamma receptor. However, although the two proteins are clearly related, they show less than 60% identity in both the putative extracellular domain and the intracellular domain.