目的构建人GMV/ARL-1真核表达载体,为进一步建立转ARL-1基因Hep-G2单克隆细胞系奠定基础.方法采用限制性内切酶技术,酶切真核骨架质粒pBK/CMV及另一质粒pBK/ARL-1(内含ARL-lcDNA全长),用TONA连结酶将目的基因连至骨架质粒上,酶切及序列分析的方法鉴定重组质粒.结果经Eco R I、Nco I、Uind Ⅲ酶切鉴定和序列分析,证明ARL-1基因已插入到pBK/CMV真核表达载体上.结论成功地构建了CHV/ARL-1真核表达载体.“,”Objective:To construct the eukaryotic expression vector CMV/ARL - 1 in order to prepare for establishing the Hep - G2 cell lines transfected with ARL - I. Methods: By endonuclease digestion, ARL - 1 gene from the plasmid pBK/ARL - 1 was cloned into the eukaryotic expression vector pBK/CMV to construct the eukaryotic expression vector CMV/ARL- I ,the recombinant plasmid was identified by endonuclease digestion and sequence analysis. Results:The ARL- I gene was identified to be inserted into the eukaryotic expression vector pBK/CMV by the digestion of Eco R Ⅰ , Nco Ⅰ , Hind Ⅲ and seqtence analysis. Conclusions: The eukaryotic expression vector CMV/ARL - 1 has been successfully constructed.