Establishment of a stable mouse FcγRIII COS7 cell line exhibiting native receptor activity

Abstract

In order to further the study the properties of the mouse Fc gamma receptor III (moFcγRIII) and mouse g chain their genes were co-expressed in cultured COS7 cell and a stable cell line was produced. The moFcγRIII and g chain genes were isolated from the mouse peripheral blood leucocyte mRNA, amplified separately by RT-PCR, and eukaryotic expression plasmids of pc3mRIII and pc3mγ were constructed. After identification by restriction digestion and PCR, the recombinant plasmids of the Pvu I-linearised pc3mRIII and the Pvu I-linearised pc3mg 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 moFcγRIII and g chain RNA was confirmed by RT-PCR, and functional expression was identified by flow cytometry and rosetting assay. The mFcγRIII expressed on the surface of selected cell line shows similar specificity. As that of native moFcγRIII and provides a novel experimental system to study further the function of moFcγRIII in a convenient replicating.© 2011 Taylor & Francis.