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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。
Objective: To construct a HEK293 cell line stably expressing human chemokine receptor 6 (CCR6). Methods: The CCR6 and Gα16 plasmids were co-transfected into HEK293 cells and HEK293 cell clones stably expressing the CCR6 gene were picked. In vitro chemotaxis assay, calcium flow assay, RT-PCR, Western blot and immunofluorescence staining were used to detect the expression of CCR6 on HEK293 cells. Results: The above experiment confirmed that the CCR6 gene and Gα16 plasmid co-transfected HEK293 cells stably expressing CCR6, and has biological activity. Conclusion: Stable expression of biologically active CCR6 on the surface of HEK293 cells lays a foundation for studying the biological function of CCR6 and screening antagonists of CCR6.