| 重组ACTH(4-10)与GDNF融合蛋白及其生物活性研究 |
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| 引用本文： | 陈哲宇,张勇,何成,路长林,吴祥甫.重组ACTH(4-10)与GDNF融合蛋白及其生物活性研究[J].生物化学与生物物理进展,2001,28(1):67-71. |
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| 作者姓名： | 陈哲宇 张勇 何成 路长林 吴祥甫 |
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| 作者单位： | 1. 第二军医大学神经生物学教研室,
2. 中国科学院上海生物化学研究所, |
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| 基金项目： | 国家自然科学基金资助项目(30000048)及军队“十五”青年基金. |
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| 摘 要： | 通过PCR方法构建了促肾上腺皮质激素4-10 (ACTH(4-10))与胶质细胞源性神经营养因子（GDNF）的融合基因，并将它重组克隆到表达载体pET-28a(+)中，构建表达质粒pET-ACTH(4-10)-GDNF，转化大肠杆菌BL21(DE3)，经IPTG诱导可高效表达ACTH(4-10)-GDNF融合蛋白.用Ni2+-NTA树脂一步法纯化目的蛋白，纯度达85％以上.纯化和复性的ACTH(4-10)-GDNF融合蛋白能显著促进脊髓神经元存活，作用强于ACTH(4-10)及GDNF蛋白.
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| 关 键 词： | 促肾上腺皮质激素4-10，胶质细胞源性神经营养因子，融合蛋白，基因表达 |
| 收稿时间： | 1999/12/4 0:00:00 |
| 修稿时间： | 1999年12月4日 |
The Recombinant ACTH(4-10)-GDNF Fusion Protein and Study of Its Biological Activity |
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| CHEN Zhe-Yu,ZHANG Yong,HE Cheng,LU Chang-Lin and WU Xiang-Fu.The Recombinant ACTH(4-10)-GDNF Fusion Protein and Study of Its Biological Activity[J].Progress In Biochemistry and Biophysics,2001,28(1):67-71. |
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| Authors: | CHEN Zhe-Yu ZHANG Yong HE Cheng LU Chang-Lin and WU Xiang-Fu |
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| Institution: | Department of Neurobiology, The Second Military Medical University, Shanghai 200433, China;Shanghai Institute of Biochemistry, The Chinese Academy of Sciences, Shanghai 200031, China;Department of Neurobiology, The Second Military Medical University, Shanghai 200433, China;Department of Neurobiology, The Second Military Medical University, Shanghai 200433, China;Department of Neurobiology, The Second Military Medical University, Shanghai 200433, China;Shanghai Institute of Biochemistry, The Chinese Academy of Sciences, Shanghai 200031, China |
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| Abstract: | The chimeric gene of ACTH(4-10) with GDNF was constructed by PCR amplification. The fused gene was inserted into the expression vector pET-28a(+) and expressed in E.coli. with a level of 30% of the total bacterial proteins. The expressed product was purified by Ni2+-NTA resin, up to 85% purity. The results of activity assays showed that the chimeric protein could significantly promote the survival of spinal cord neurons and had a higher neurotrophic activity than ACTH(4-10) and GDNF respectively. |
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| Keywords: | adrenocorticotropin(4-10) glial cell line derived neurotrophic factor chimeric protein gene expression |
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