中文摘要：

目的：构建人DC-SIGN绿色荧光融合蛋白的真核表达载体,在COS7细胞表面表达鉴定。方法：用RT-PCR从人外周血单个核细胞扩增DC-SIGN蛋白基因片段,连接到T载体。测序鉴定后切下相应片段,连接到pEGFP-C1表达载体,重组质粒（命名为：DS-pEGFP-C1）转染COS7细胞,表达DC-SIGN绿色荧光融合蛋白（命名为：DS-EGFP）。荧光定量PCR检测融合基因mRNA拷贝量,激光扫描共焦显微镜鉴定DS-pEGFP-C1的表达和摄取减毒牛型分枝杆菌BCG功能。结果：用RT-PCR扩增得到正确的目的基因片段并构建成真核表达载体DS-pEGFP-C1,转染COS7细胞,荧光定量PCR检测DS-pEGFP-C1转录mR-NA拷贝量是4.52×10^11 copies/ml,激光扫描共焦显微镜检测证实DS-EGFP在COS7细胞表面表达,同时具有摄取BCG的功能。结论：本研究成功构建人DC-SIGN绿色荧光融合蛋白的真核表达载体DS-pEGFP-C1,表达了融合蛋白,后者能够摄取BCG。

英文摘要：

Objective：To construct the eukaryotic vector of human DC-SIGN and EGFP fusion protein, and to identify the protein in cell line COS7. Methods： RT-PCR, T-vector and pEGFP-C1 vector were used to construct the recombinant expressing plasmid encoding for the fusion protein of DC-SIGN and EGFP. COS7 cells were transfected with the plasmid. Real-time PCR and laser scanning confocal microscope were used to quantificate expression of the fusion protein and cell function of uptaking BCG. Results： DC-SIGN eDNA was successfully cloned into the eukaryotic vector pEGFP-C1. The recombinant vector was transfected into COS7, real-time PCR test showed the amount of mRNA encoding for the fusion protein was 4.52 × 10^11 copies/ml and laser scanning confocal nficmscope confirmed that the cells could uptake BCG. Conclusion： We have constructed a recombinant vector expressing DC-SIGN and EGFP fusion protein.