人趋化因子受体CCR6在HEK293细胞内的稳定表达及其功能分析

Expression and function analysis of chemokine receptor CCR6 in HEK293 cells

目的:构建稳定表达人趋化因子受体6(CCR6)的HEK293细胞株。方法:将CCR6基因和Gα16质粒共转到HEK293细胞中,并挑取稳定表达CCR6基因的HEK293细胞克隆。采用体外趋化实验、钙流实验、RT-PCR、Western blot及免疫荧光染色法,检测CCR6在HEK293细胞表面的表达。结果:经上述实验证实,CCR6基因和Gα16质粒共转染的HEK293细胞上,可稳定表达CCR6,且具有生物学活性。结论:成功地在HEK293细胞表面稳定表达具有生物学活性的CCR6,为研究CCR6的生物学功能及筛选CCR6的拮抗剂奠定了基础。

AIM： To construct a cell line which stably expresses human chemokine receptor 6 （CCR6）. METHODS： The human CCR6 cDNA and plasmid G were cotransfected into HEK 293 cells and the clones stably expressing CCR6 were picked out. The expression of CCR5 in HEK293 cells was detected by RTPCR, Western blotting, immunofluorescence test, calcium mobilization and in vitro chemotaxis assay.