关键词: granzyme B;基因表达;HeLa细胞

    摘 要:目的 构建携带人granzyme B基因的真核表达载体，并将其在HeLa细胞中表达. 方法 用反转录PCR获取人granzyme B全长cDNA序列，将其活性型序列克隆进pcD-NA3重组表达质粒.脂质体法转染HeLa细胞，间接免疫荧光检测目的蛋白表达，HE染色观察其对转染HeLa细胞形态的影响. 结果 成功获得了野生型granzyme B cDNA，构建了编码活性型granzyme B的真核表达载体pcDNA3-G.转染HeLa细胞后观察到目的蛋白表达，转染细胞形态发生变化，出现多核大细胞及固缩小细胞. 结论 活性型granzyme B哺乳动物表达系统的建立，为进一步研究granzyme B的生物学效应奠定了基础.

    Expression of human granzyme B gene in HeLa cells

    ZHAO Jing XU Yan-Ming JIA Lin-Tao YU Cui-Juan CHEN Lei HUANG Wei-Quan WANG Cheng-Ji

    YANG An-Gang

    1 Department of Biochemistry and Molecular Biolo-gy，

    2 Department of Histology and Embryology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China

    Keywords:granzyme B;gene expression;HeLa cells

    Abstract:AIM To construct eukaryotic expression vector carrying human granzyme B gene and express it in HeLa cells.METHODS RT-PCR was used to obtain human granzyme B cDNA.Then the recombinant expression vector pcDNA3-G was constructed by cloning active ganzyme B into eukaryotic expression vector pcDNA3.After transfection into HeLa cells through lipofectamine2000 ......