| ISOLATION,CHARACTERIZATION AND EXPRESSION OF THE COMPLEMENTARY DNA FOR HUMAN TUMOR NECROSIS FACTOR (TNF-α) |
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| 引用本文： | 肖蕾,田园,王秀琴,贾凤兰,吴旻.ISOLATION,CHARACTERIZATION AND EXPRESSION OF THE COMPLEMENTARY DNA FOR HUMAN TUMOR NECROSIS FACTOR (TNF-α)[J].中国科学B辑(英文版),1990(10). |
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| 作者姓名： | 肖蕾 田园 王秀琴 贾凤兰 吴旻 |
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| 作者单位： | National Laboratory of Molecular Oncology，Cancer Institute，Chinese Academy of Medical Sciences，Beijing 100021，PRC，National Laboratory of Molecular Oncology，Cancer Institute，Chinese Academy of Medical Sciences，Beijing 100021，PRC，National Laboratory of Molecular Oncology，Cancer Institute，Chinese Academy of Medical Sciences，Beijing 100021，PRC，Insitute of Hepartitis，Beijing Army General Hospital，Beijing，National Laboratory of Molecular Oncology，Cancer Institute，Chinese Academy of Medical Sciences，Beijing 100021，PRC |
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| 摘 要： | A cDNA for human TNF-α (615bp) was isolated by means of polymerase chain reaction (PCR) using first strand cDNA from PMA-induced HL-60 cells as template. The result from sequencing the 615 bp cDNA fragment indicated that it corresponded to the entire sequence of mature human TNF coding region. Direct expression of mature human TNF was achieved using a plasmid pHT-1 constructed by ligation of the cDNA and a synthetic DNA. The IPTG-induced bacterial product (hTNF) showed cytotoxicity to mouse L-929 cells. The TNF activity was further identified by neutralization of a specific monoclonal antibody against human TNF-α. Approximately 80,000 units of activity were detected per ml of culture at A600=2.
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