moFcγRⅢ、γ链真核表达载体的构建及其共转染稳定细胞系的建立

Establishment of a Cell Line Cotransfected with a Eukaryotic Expression Vector Stably Expressing moFcγRⅢ and γ Chain

将moFcγRⅢ、γ链编码区cDNA分别亚克隆到真核表达载体pcDNA3的巨细胞病毒启动子下游,构建了重组表达质粒pc3moRⅢ、pc3moγ;用PvuⅠ线性化重组质粒,以线性化重组质粒共转染COS-7细胞,用玫瑰花环试验检测moFcγRⅢ在转染细胞表面的表达,通过G418抗性筛选和单细胞克隆,在转染细胞表面稳定表达了moFcγRⅢ受体分子。稳定转染细胞系的建立和moFcγRⅢ受体分子基因的表达,为进一步研究moFcγRⅢ的功能提供了基础。

The cDNA for the complete encoding region of moFcγRⅢ and γ-chain were separately cloned into the mammalian expression vector pcDNA3 to construct expressing plasmids,pc3moRⅢand pc3moγ.COS-7 cells were cotransfected with the linear expressing plasmids,selected by G418 and detected by rosette assay for the expression of moFcγRⅢ.The establishment of the stably-transfected cell line expressing target gene provides a basis for further studies on the function of the moFcγRⅢ gene.