| Construction of shuttle, expression vector of human tumor necrosis factor alpha (hTNF-α) gene and its expression in a cyanobacterium, Anabaena sp. PCC 7120 |
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| 引用本文： | 刘凤龙
,施定基
,商之狄
,邵宁
,徐旭东
,钟泽璞
,张宏斌
,吴锦银
,王捷
,江悦华
,赵树进
,林晨
,张雪艳
,吴旻
,彭国宏
,张海霞
,曾呈奎.Construction of shuttle, expression vector of human tumor necrosis factor alpha (hTNF-α) gene and its expression in a cyanobacterium, Anabaena sp. PCC 7120[J].中国科学：生命科学英文版,1999(1). |
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| 作者姓名： | 刘凤龙 施定基 商之狄 邵宁 徐旭东 钟泽璞 张宏斌 吴锦银 王捷 江悦华 赵树进 林晨 张雪艳 吴旻 彭国宏 张海霞 曾呈奎 |
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| 作者单位： | 1. Laboratory of Photosynthesis，Institute of Botany，Chinese Academy of Sciences，Beijing 100093，China;
2. The Guangzhou General Hospital of Guangzhou Military Command，Guangzhou 510010，China;
3. StateKey Laboratory of Molecular Oncology，Institute of Oncology，Chinese Academy of Medicine，Beijing 100021，China;
4. EMBL，Institute of Oceanology，Chinese Academy of Sciences，Qingdao 266071，China |
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| 基金项目： | Project supported by the National Natural Science Foundation of China (Grant No. 39280016). |
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| 摘 要： | The construction of the shuttle, expression vector of human tumor necrosis factor alpha (hTNF-a) gene and its expression in a cyanobacterium Anabaena sp. PCC 7120 was reported. The 700-bp hTNF cDNA fragments have been recovered from plasmid pRL-rhTNF, then inserted downstream of the promoter PpsbA in the plasmid pRL439. The resultant intermediary plasmid pRL-TC has further been combined with the shuttle vector pDC-8 to get the shuttle, expression vector pDC-TNF. The expression of the rhTNF gene in Escherichia coli has been analyzed by SDS-PAGE and thin-layer scanning, and the results show that the expressed TNF protein with these two vectors is 16.9 percent (pRL-TC) and 15.0 percent (pDC-TNF) of the total proteins in the cells, respectively, while the expression level of TNF gene in plasmid pRL-rhTNF is only 11.8 percent. Combined with the participation of the conjugal and helper plasmids, pDC-TNF has been introduced into Anabaena sp PCC 7120 by triparental conjugative transfer, and the stable transgenic
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| 关 键 词： | human tumor necrosis factor alpha (hTNF-α) Anabaena sp. PCC 7120 shuttle expression vector triparental conjugative transfer Southern and Western blottings cytotoxicity. |
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