| 果蝇血液发育标记基因mrj的多克隆抗体制备及检测 |
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| 引用本文： | 雷旻音,杨哲,黄婷,刘丹,李永青,吴秀山.果蝇血液发育标记基因mrj的多克隆抗体制备及检测[J].湖南师范大学自然科学学报,2012(4):55-58. |
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| 作者姓名： | 雷旻音 杨哲 黄婷 刘丹 李永青 吴秀山 |
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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 |
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| 基金项目： | 国家自然科学基金资助项目(30671053,30871340,30930054) |
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| 摘 要： | mrj是黑腹果蝇中一个蛋白编码基因.根据已报道的mrj基因序列,利用生物信息学分析选取果蝇mrj基因抗原亲水区,通过PCR扩增出其部分编码区序列,将其连接到pET-28a原核表达载体上.经酶切及测序鉴定质粒构建成功后,将重组质粒(pET-28a-mrj)转入Rosetta菌后通过IPTG(Isopropy1β-D-thiogalactoside)诱导表达出带His标签的重组融合蛋白后用镍柱纯化.将纯化的蛋白免疫新西兰大白兔制备多克隆抗体,并用Western Blot检测抗体的效价和特异性.
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| 关 键 词： | mrj基因 果蝇 多克隆抗体 |
Preparation and Assay of Polyclonal Antibody of Blood Development Marker Gene Mrj in Drosophila Melanogaster |
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| LEI Min-yin,YANG Zhe,HUANG Ting,LIU Dan,LI Yong-qing,WU Xiu-shan.Preparation and Assay of Polyclonal Antibody of Blood Development Marker Gene Mrj in Drosophila Melanogaster[J].Journal of Natural Science of Hunan Normal University,2012(4):55-58. |
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| Authors: | LEI Min-yin YANG Zhe HUANG Ting LIU Dan LI Yong-qing WU Xiu-shan |
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| Institution: | (Center for Heart Development,Key Lab of MOE for Development Biology and Protein Chemistry, Hunan Normal University,Changsha 410081,China) |
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| Abstract: | Mrj is a protein coding gene from Drosophila.Hydrophilic and well specific fragment of mrj is selected by bioinformatics method.Then it was amplified the fragment by PCR and cloned into the expression vector pET-28a to acquire the recombinant expression plasmid pET-28a-mrj.It was transformed into Escherichia coli Rosetta,and the fusion protein with His tag was induced with IPTG.The fusion protein was purified by nickel chelating resin.His-mrj fusion protein was used to immune the New Zealand white rabbits,then identified the antibody by Western blot. |
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| Keywords: | mrj gene Drosophila polyclonal antibody |
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