关键词: 肿瘤坏死因子;hPgnK5;内皮，血管;抑制因子，免疫

    摘 要:目的 获得hTNFα-hPgnK5融合蛋白基因，在原核细胞中表达融合蛋白，探索其活性. 方法 PCR法改造hTNFα基因，消除其终止密码子，构建hTNFα-hPgnK5融合蛋白基因.在大肠杆菌DH5α中表达该融合蛋白，测定其体外抑制肿瘤细胞或血管内皮细胞增殖的活性. 结果 hTNFα基因经PCR法改造，去除了终止密码子，并在3'端引入BamHI位点.hTNFα-hPgnK5融合蛋白基因构建完成，并在原核系统中获得表达，免疫印迹反应表明该融合蛋白具有hPgnK5免疫活性.体外实验表明该融合蛋白可抑制肿瘤细胞和血管内皮细胞的增殖. 结论 hPgnK5蛋白可与hTNFα一起表达，该融合蛋白有抑制肿瘤细胞和血管内皮细胞增殖的活性.

    Expression and characterization of hTNFα┐hPgnK5fusion protein in prokaryotic system

    FAN Bin YAN Zhen ZHANG Ying-Qi ZHAO Ning YAO Li-Bo Su Cheng-Zhi

    1 Biotechnology Centre，

    2 Department of Biochem-istry&Molecular Biology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China

    Keywords:tumor necrosis factor;hPgnK5;endothelium，vasculav;suppressor factors，immundogic

    Abstract:AIM To obtain the gene of hTNFα-hPgnK5fu-sion protein，express the fusion protein in prokaryotic system and research its characterization.METHODS hTNFαgene was reformed by PCR and its termination codon eliminated，the hTNFα-hPgnK5fusion protein gene then constructed was and the fusion protein in prokaryotic system expressed ......