Establishment of a stable mouse Fc gamma RIII COS7 cell line exhibiting native receptor activity

文献类型： 外文期刊

第一作者： Xi, J.

作者： Xi, J.;Zhang, L. N.;Wang, L.;Miao, X. W.;Liu, Y. C.;Tian, X. H.;Deng, R. G.;Xi, J.;Bu, G. H.;Lu, Q. Y.

作者机构：

关键词： moFc gamma RIII and gamma chain;co-transfection;flow cytometry;rosetting assay

期刊名称：JOURNAL OF APPLIED ANIMAL RESEARCH （ 影响因子：1.63； 五年影响因子：1.727 ）

ISSN： 0971-2119

年卷期： 2011 年 39 卷 2 期

页码：

收录情况： SCI

摘要： In order to further the study the properties of the mouse Fc gamma receptor III (moFc gamma RIII) and mouse gamma chain their genes were co-expressed in cultured COS7 cell and a stable cell line was produced. The moFc gamma RIII and gamma chain genes were isolated from the mouse peripheral blood leucocyte mRNA, amplified separately by RT-PCR, and eukaryotic expression plasmids of pc3mRIII and pc3m gamma were constructed. After identification by restriction digestion and PCR, the recombinant plasmids of the Pvu I-linearised pc3mRIII and the Pvu I-linearised pc3m gamma were co-transfected into COS7 cells with lipofectamine. Cloning by limiting dilution after selective culture with G418, established a stably-transfected cell line successfully. Co-expression of the moFc gamma RIII and gamma chain RNA was confirmed by RT-PCR, and functional expression was identified by flow cytometry and rosetting assay. The moFc gamma RIII expressed on the surface of selected cell line shows similar specificity. As that of native moFc gamma RIII and provides a novel experimental system to study further the function of moFc gamma RIII in a convenient replicating cell line.

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