中文摘要：

目的：构建真核表达重组体PcDNA3．1-BEGγ的稳定转染细胞株，为进一步深入研究REGγ的功能奠定基础。方法：提取MCF-γ细胞的总RNA为模板，通过RT—PCR获取全长REGγcDNA，经限制性内切酶EcoRI、EcoRV酶切后与PcDNA3．1连接构建重组体，再经内切酶酶切及DNA序列分析鉴定重组体构建正确。以Lipokctamine2000将重组体导入HBL-100细胞，由G418筛选出稳定转染细胞株。结果：经免疫细胞化学和RT—PCR检测并证实稳定转染细胞株中有REG1的高表达。结论：真核表达重组体PcDNA3．1-REGγ构建成功，并经抗生素筛选获得了稳定高表达REGγ的细胞株，为进一步研究REGγ的功能奠定了基础。

英文摘要：

Objective： To construct the stable transfective cell line with the eukaryotic expression vector for human REGγ cDNA to laid the foundation for further study of the function of REG γ. Methods： REG γ, cDNA was obtained by RT-PCR of total RNA extracted from the human breast cancer cell MCF-7, and it was digested by the restriction endonuclease EcoR I and EcoR V before connection with PcDNA3.1.The recombination with forward insert were slelected by restriction endonuclease digestion and confirmed correct by sequencing and then transfected into HBL-100 cell by using lipofectamine2000.The stable transfeeted cell line was selected in medium containing antibiotic （G418）. Results： The result of immunocytochemistry and RT-PCR of total RNA extracted from the stable transfected cell line showed that there existed the overexpression of REG 5＇ cDNA in it. Conclusion： The construction of the eukaryotic expression vector for REG 5＇ was successful, and the stable transfected cell line for overexpression REG γ was obtained by G418 selection. This work may pave the way for further study of the functions of REGγ.