| GST-HRB融合蛋白的表达与纯化 |
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| 引用本文： | 刘丽杰,高学娟,刘小会,陈妙娟,朱森,刘朗夏.GST-HRB融合蛋白的表达与纯化[J].生物技术通报,2011(11). |
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| 作者姓名： | 刘丽杰 高学娟 刘小会 陈妙娟 朱森 刘朗夏 |
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| 作者单位： | 暨南大学生命科学技术学院,广州,510632 |
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| 基金项目： | 广东省自然科学基金项目(9151065004000005); 中央高校基本科研业务费专项资金资助项目(21609705,21609317) |
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| 摘 要： | 构建GST-HRB重组质粒,进行融合蛋白的表达、纯化及鉴定.利用PCR扩增及基因重组技术,以pcDNA-3.1-HRB为模板扩增出HRB全基因序列,并将其插入带有GST(谷胱甘肽巯基转移酶)标签的原核表达载体pGEX-6P-1中,构建GST-HRB融合蛋白表达质粒.然后,将重组质粒GST-HRB转化至大肠杆菌Rosseta进行融合蛋白的表达.利用GST琼脂糖珠进行融合蛋白的纯化,最后应用SDS-PAGE电泳和Western blotting鉴定纯化的融合蛋白.结果表明,成功构建pGEX-6P-1-HRB原核表达载体,表达及纯化了GST-HRB融合蛋白.
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| 关 键 词： | GST-HRB 构建 融合蛋白 表达 纯化 |
The Expression and Purification of GST-HRB Fusion Protein |
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| Liu Lijie,Gao Xuejuan,Liu Xiaohui,Chen Miaojuan,Zhu Sen,Liu Langxia.The Expression and Purification of GST-HRB Fusion Protein[J].Biotechnology Bulletin,2011(11). |
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| Authors: | Liu Lijie Gao Xuejuan Liu Xiaohui Chen Miaojuan Zhu Sen Liu Langxia |
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| Institution: | Liu Lijie Gao Xuejuan Liu Xiaohui Chen Miaojuan Zhu Sen Liu Langxia(College of Life Science and Technology,Jinan University,Guangzhou 510632) |
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| Abstract: | It was to construct a GST-HRB recombinant plasmid,and identify fusion protein.Using PCR and recombinant DNA technology,HRB full gene sequence was amplified from template pcDNA-3.1-HRB and inserted into the prokaryotic expression vector pGEX-6P-1 with tag GST(glutathione sulfurtransferase).The recombinant vector was then transformed into the E.coli Rosetta strain to produce the GST-HRB fusion protein,which was purified through a Sepharose-Glutathione column,and verified by SDS-PAGE and Western blotting using... |
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| Keywords: | GST-HRB Construction Fusion protein Expression Purification |
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