关键词: 肝炎病毒，乙型;酵母菌;杂交;基因，报告

    摘 要:目的 用含乙型肝炎病毒(HBV)前S区不同大小的cDNA片段构建酵母双杂交诱饵载体，并检测其表达产物对酵母细胞有无毒性作用及对报告基因有无激活作用. 方法 PCR扩增含HBV前S区不同大小的cDNA片段，分别克隆入pUC19质粒，经测序正确后，再分别亚克隆入酵母双杂交诱饵载体pGBKT7中.将重组质粒导入酵母菌AH109，检测其表达产物在酵母细胞中对报告基因有无激活作用. 结果 成功获得含HBV前S区不同大小的cDNA片段，HBV前S区不同片段所表达的蛋白对酵母菌AH109无毒性，但是对报告基因均有激活作用. 结论 不能利用酵母双杂交Gal4系统3来研究与HBV前S区相互作用的蛋白;证实了HBV前S区不同片段所表达的蛋白的自激活功能，为进一步研究乙型肝炎病毒致病机制打下了基础.

    Reporter genes assay of the different fragments of HBV preS region in yeast two┐hybrid system

    ZHANG Wei LIU Xin-Ping ZHANG Jing YAO Li-Bo CHEN Chang-Qing

    1 Department of Biochemistry and Molecular Biolo-gy，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China，

    2 Centre of Shanghai Biology Engineering，Academy of Sci-ences of China，Shanghai200032，China

    Keywords:hepatitis B virus;yeasts;hybridization;genes，reporter

    Abstract:AIM To construct the bait vector with different sizes of HBV PreS region in yeast two-hybrid system，then assay whether its expression product can affect the growth of yeast cell and activate the reporter genes.METHODS The cDNA fragments encoding different sizes of HBV PreS region were amplified by PCR ......