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重组ACTH(4-10)与GDNF融合蛋白及其生物活性研究
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
作者姓名:陈哲宇  张勇  何成  路长林  吴祥甫
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,
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.
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.

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:1999-12-04
修稿时间:1999-12-04

The Recombinant ACTH(4-10)-GDNF Fusion Protein and Study of Its Biological Activity
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
Authors:CHEN Zhe-Yu  ZHANG Yong  HE Cheng  LU Chang-Lin  WU Xiang-Fu
Affiliation: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
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.
Keywords:adrenocorticotropin(4-10)   glial cell line derived neurotrophic factor   chimeric protein   gene expression
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