田耕,易继林,华中科技大学同济医学院附属同济医院普外科 湖北省武汉市 430030

    田耕,男, 1971-03-18生,陕西省宝鸡市人,汉族, 博士,主治医师.

    项目负责人:田耕, 430030, 湖北省武汉市汉口解放大道1095号,华中科技大学同济医学院附属同济医院普外科. geng_tian707@hotmail.com

    电话:027-83663402

    收稿日期:2003-03-08 接受日期:2003-03-25

    Genecloning of murine a-fetoproteingene and construction of its eukaryotic expression vector and expressionin CHO cells

    GengTian, Ji-Lin Yi

    GengTian, Ji-Lin Yi, Department of General Surgery, Tongji Hospital, TongjiMedical College, Huazhong University of Science and Technology,Wuhan430030, Hubei Province, China

    Correspondence to: Geng Tian, Department of General Surgery ,Tongji Hospital, Tongji Medical College, Huazhong University of Scienceand Technology, Wuhan 430030, Hubei Province, China. geng_tian707@hotmail.com

    Received:2003-03-08 Accepted:2003-03-25

    AbstractAIM: To clone murine AFP gene, to construct the eukaryotic expressionvector of AFP and express it in CHO cells.

    METHODS:Total RNA was extracted from Hepa1-6 cells, then the murine AFP gene wasamplified by RT-PCR and cloned into the eukaryotic expression vectorpcDNA3.1. The recombinant vector was transformed into E.coli. DH5a,the positive clones were selected and plasmid DNA was identified byrestriction enzyme analysis and sequencing. After transient transfectionof CHO-K1 cells with the recombinant vector, Western blotting was used todetect the expression of AFP. ......