趋化因子受体CCR6的克隆及其在HEK293细胞中的表达

目的：构建人趋化因子受体6（CCR6）cDNA序列的真核表达载体，并了解其在HEK293细胞中的表达。方法：提取人淋巴结总RNA，通过逆转录PCR扩增出CCR6基因片段，并构建真核表达载体pcDNA3，1（＋）-CCR6；重组载体通过脂质体转染HEK293细胞，免疫荧光法鉴定CCR6的表达。结果：酶切鉴定和序列分析证实重组质粒含有CCR6编码序列．转染实验表明重组质粒能在HEK293细胞中表达出具有活性的CCR6片段。结论：CCR6真核表达载体构建及表达成功，为下一步CCR6拈抗剂的筛选奠定了基础。

Construction of eukaryotic expression vectors containing coding of human chemokine receptor 6(CCR6) gene and its expression in HEK293 cells

Objective:To construct an eukaryotic expression vector containing the coding region of human chemokine recptor 6(CCR6) gene and to detect its expression in HEK293 cells.Methods:The CCR6 gene was cloned into eukaryotic expression vector pcDNA3.1(+),The recombinant pcDNA3.1(+)-CCR6 was transfected into HEK293 cells by DOTAP Lipofectamine Reagent.Immunoflourescence and FCM were used to dctect the CCR6 gene expression in HEK293 cells.Results:Restriction analysis and DNA sequencing showed that the recombinant Plasmid contained the coding region of human chemokine recptor 6(CCR6) gene.Transfection experiment verified that CCR6 gene could be expressed in HEK293 cells.Conclusion:The eukaryotic expression vector containing the CCR6 gene was succesfully constructed and expressed,which is a basis for the study for CCR6 antagonists.