| 人胶质细胞源性神经营养因子基因片段克隆及其在大肠杆菌中的表达 |
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| 引用本文： | 刘亚珍,施雨露,王友联,刘永茂,万全. 人胶质细胞源性神经营养因子基因片段克隆及其在大肠杆菌中的表达[J]. 国际生物制品学杂志, 2011, 34(1): 1-4. DOI: 10.3760/cma.j.issn.1673-4211.2011.01.001 |
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| 作者姓名： | 刘亚珍 施雨露 王友联 刘永茂 万全 |
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| 作者单位： | 130021长春, 吉林大学第一医院药剂科（刘亚珍）；130021长春, 吉林大学白求恩医学院实验中心（施雨露、刘永茂）；130051长春, 白求恩医科大学制药厂（王友联）；130051 长春, 吉林大学生命科学学院08级生物技术系（万全） |
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| 摘 要： |
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| 关 键 词： | 神经生长因子类 基因表达 逆转录聚合酶链反应 |
Gene cloning of human glial cell line-derived neurotrophic factor and its expression in E. coli |
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| LIU Ya-zhen,SHI Yu-lu,WANG You-lian,LIU Yong-mao,WAN Quan. Gene cloning of human glial cell line-derived neurotrophic factor and its expression in E. coli[J]. International Journal of Biologicals, 2011, 34(1): 1-4. DOI: 10.3760/cma.j.issn.1673-4211.2011.01.001 |
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| Authors: | LIU Ya-zhen SHI Yu-lu WANG You-lian LIU Yong-mao WAN Quan |
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| Affiliation: | *Department of Pharmaceutics, The First Hospital of Jilin University, Changchun 130021, China |
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| Abstract: | Objective To express human glial cell line-derived neurotrophic factor (GDNF)in E. coli (BL21). Methods Gene segment of GDNF was amplified using the genomic DNA from human glioma as template by polymerase chain reaction, and was directly ligated with vector pET28a+. The recombinant expression vector pET28a+ -GDNF was transformed into E. coli BL21 ( DE3 ) and induced with isopropyl β-D-thiogalactoside. The expression product was identified by SDS-PAGE after gel chromatography isolation.Results Gene segment of pET28a+ -GDNF was identified as the sequence of encoding mature peptide of GDNF by restriction analysis and DNA sequencing. SDS-PAGE revealed that the purified product had a M.16 000protein band. Conclusion The prokaryotic expression system we constructed can express human GDNF. |
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| Keywords: | Nerve growth factors Gene expression RT-PCR |
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