| 人β神经生长因子在大肠杆菌中的高表达 |
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| 引用本文： | 张剑 张林元. 人β神经生长因子在大肠杆菌中的高表达[J]. 微生物学报, 1997, 37(6): 429-433 |
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| 作者姓名： | 张剑 张林元 |
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| 作者单位： | 南京师范大学生物系，南京师范大学生物系，南京师范大学生物系，南京军事医学研究所分子遗传室，复旦大学国家重点实验室 南京 210097，南京 210097，南京 210097 |
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| 摘 要： | 将编码人β神经生长因子(Huβ-HGF)的基因克隆到由T7噬菌体启动子控制的pET11c大肠杆菌表达载体中,重组质粒经鉴定含有Huβ-NGF基因,未解聚的表达产物经SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE),结果显示出二聚体27kD的蛋白带。而完全解聚的表达产物SDS-PAGE显示出一条13β5kD单体带。经凝胶电泳扫描,表达带占菌体总蛋白的14.5％。用兔抗鼠β—NGF的多克隆抗体进行的Western-Blot的结果表明,二聚体同单体都有免疫原性。在生物活性的鉴定中,菌体表达产物可以使小鸡鸡胚的背根神经节产生神经元突起,由此可以证明该表达产物有较高的生物活性。
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| 关 键 词： | 人β申经生长因子 Huβ-NGF基因 基因表达 rHuβ-NGF 鸡胚背根神经节 |
The high expression of human beta-nerve growth factor in E. coli] |
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| Zhang Jian Zhang Lingyuan Zhang Xiran Chai Jianhua Li Chaojun. The high expression of human beta-nerve growth factor in E. coli][J]. Acta microbiologica Sinica, 1997, 37(6): 429-433 |
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| Authors: | Zhang Jian Zhang Lingyuan Zhang Xiran Chai Jianhua Li Chaojun |
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| Affiliation: | Deartment of Biology, Nanjing Normal University, Nanjing 210097. |
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| Abstract: | The Hu beta-NGF gene encoding the beta subunit of mature human nerve growth factor was cloned into the pET11c vector under the control of T7 bacteriophage promoter. The SDS-PAGE electrophoresis results of nonreduced recombinant products showed a clear band corresponding to homodimeric (27 kD) form of the molecule. But the SDS-PAGE results of reduced expression protein showed a single band corresponding to monomeric form of Hu beta-NGF (13.5 kD). It represented approximately 14.5% of the total cellular protein. Both of them were immunopositive on Western-Blots with rabbit anti-m beta-NGF polyclonic antibodies. And the recombinant product was biologically active on cultured chicken dorsal root ganglion neurons. So it demonstrated the feasibility of synthesizing the biologically active forms of Hu beta-NGF in E. coli. |
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| Keywords: | Hup-NGF Hup-NGF gene Gene expression rHup-NGF Chicken dorsal root ganglion neurons |
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