摘要： 为建立小鼠IgGFc受体(moFcγRⅡ)稳定表达的哺乳动物细胞系,PCR克隆小鼠moFcγRⅡ片段,构建真核表达载体pcDNAmoFcγRⅡ。转染COS-7细胞后通过G418抗性筛选和连续克隆,获得了在COS-7细胞表面稳定表达FcγRⅡ的真核细胞系。玫瑰花环试验结果显示其阳性率在90%以上。
关键词:
小鼠IgG受体(Fc&gamma,
RⅡ),
细胞系,
玫瑰花环,
稳定转染
Abstract: To establish the mouse IgGFc Receptor(moFcγRII)stable expression cell line.The coding sequence of FcγRII was amplified and cloned into the mammalian expression vector pcDNA3.0 under control of the CMV promoter.The COS-7 cells were tranfected with the recombinant plasmids,and the FcγRII stable expression COS-7 cells were selected by the antibiotics of G418,and subcloned by well-known method of limiting dilution.Rosetting test of the binding of mouse IgG-sensitized chicken erythrocytes show that the percent of positive cells were more than 90%.The establishment of the mouse FcγRII stable expression cell line laid the necessary foundation for the identification of the linear epitope of IgG binding on mouse FcγRII,and further investing the immunologic mechanism of Fc receptor of IgG.
Key words:
Mouse FcRII,
Cell line,
Rosetting test,
Stable transfection
中图分类号:
苗现伟, 席俊, 刘玺, 张改平, 邓瑞广, 李清州, 张利娜, 游雷鸣, 刘运超. 小鼠IgGFc受体(moFcγRⅡ)稳定表达细胞系的建立及鉴定[J]. 华北农学报, 2010, 25(2): 133-135. doi: 10.7668/hbnxb.2010.02.026.
MIAO Xian-wei, XI Jun, LIU Xi, ZHANG Gai-ping, DENG Rui-guang, LI Qing-zhou, ZHANG Li-na, YOU Lei-ming, LIU Yun-chao. Establishment of Cell Line Stably Transfected with the Constructed Plasmid of Mus Musculus FcγRⅡ[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(2): 133-135. doi: 10.7668/hbnxb.2010.02.026.