| 斑马鱼Nfatc3基因多克隆抗体的制备及检测 |
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| 引用本文： | 冯德峰,屈永贵,袁佳佳,陈发,吴秀山,李永青. 斑马鱼Nfatc3基因多克隆抗体的制备及检测[J]. 湖南理工学院学报：自然科学版, 2014, 0(2): 21-24 |
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| 作者姓名： | 冯德峰 屈永贵 袁佳佳 陈发 吴秀山 李永青 |
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| 作者单位： | 湖南师范大学 蛋白质化学及鱼类发育生物学教育部重点实验室,心脏发育研究中心 湖南省生物发育工程及新产品研发协同创新中心,长沙 410081 |
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| 基金项目： | 国家自然科学基金资助项目（81270156）;湖南省社会发展支撑计划项目（2014SK3099） |
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| 摘 要： | 为了在斑马鱼模型中更深入地研究Nfatc3蛋白在心脏发育中的功能，采用PCR技术扩增斑马鱼Nfatc3基因的部分编码区并插入pET-28a表达载体中，之后将重组质粒转入大肠杆菌（E.coli）通过IPTG诱导表达His-Nfatc3融合蛋白，对该融合蛋白采用Ni-IDA凝胶柱层析纯化后，免疫新西兰兔制备了多克隆抗体，并用western blotting对抗体进行分析.结果表明获得了高效价的特异性兔抗Nfatc3多克隆抗体.这为Nfatc3功能的进一步研究奠定了基础.
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| 关 键 词： | Nfatc3 斑马鱼 融合蛋白 多克隆抗体 |
Preparation and Detection of Polyclonal Antiserum of Nfatc3Gene in Zebrafish |
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| FENG De-feng,QU Yong-gui,YUAN Jia-jia,CHEN Fa,WU Xiu-shan,LI Yong-qing. Preparation and Detection of Polyclonal Antiserum of Nfatc3Gene in Zebrafish[J]. Journal of Hunan Institute of Science and Technology, 2014, 0(2): 21-24 |
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| Authors: | FENG De-feng QU Yong-gui YUAN Jia-jia CHEN Fa WU Xiu-shan LI Yong-qing |
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| Affiliation: | (The Center for Heart Development, Key Lab of MOE for Development Biology and Protein Chemistry, Hunan Normal University; The Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province, Changsha 41008 l, China) |
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| Abstract: | To better understand the function of theNfatc3 protein in heart development by using zebrafish as a model, here, a fraction of the encoded region in the zebrafishNfatc3was obtained by PCR amplification, then the fragment was inserted into pET-28a vector to establish the expressing system. The recombinant plasmid was transformed into E.coli and fusion protein was induced by IPTG. The purified protein was obtained by treating New Zealand white rabbits to prepare antibody. The antibody was assayed by Western blotting. The result shows that the polyclonal antibody is of high sensitivity and specificity, which lays a solid foundation for the further studies ofNfatc3function. |
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| Keywords: | Nfatc3 zebrafish fusion protein polyclonal antiserum |
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