| LRP16融合蛋白载体的构建及其原核表达 |
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| 引用本文： | 孟元光,韩为东,黄柯,李琦,伍志强,宋磊. LRP16融合蛋白载体的构建及其原核表达[J]. 中国肿瘤临床与康复, 2006, 13(3): 212-214 |
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| 作者姓名： | 孟元光 韩为东 黄柯 李琦 伍志强 宋磊 |
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| 作者单位： | 1. 解放军总医院妇产科,北京,100853
2. 解放军总医院基础所分子生物室 |
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| 基金项目： | 国家自然科学基金(编号30471813)，北京市自然科学基金(编号7052061)资助项目 |
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| 摘 要： | 目的观察和鉴定LRP16在大肠杆菌中的表达,并分离、纯化其蛋白产物。方法应用DNA重组技术,用RT-PCR方法从正常人血液中扩增出LRP16基因全长及其抗原表位区,构建重组表达质粒pRSET-C-LRP16,IPTG诱导表达并纯化融合蛋白采用SDS-PAGE凝胶电泳进行鉴定。结果成功构建LRP16基因融合蛋白载体,SDS-PAGE显示pRSET-C-LRP16表达于原核细胞,蛋白经电泳分离,切胶后获得纯化的LRP16蛋白。结论制备的高纯度的LRP16蛋白,可在大肠杆菌中稳定表达。
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| 关 键 词： | LRP16基因 融合蛋白 |
| 文章编号： | 1005-8664(2006)03-0212-03 |
| 修稿时间： | 2005-12-20 |
Construction of human LRP16 fusion protein vector and its expression in E.coli |
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| MENG Yuan-guang,HAN Wei-dong,HUANG Ke,et al. Construction of human LRP16 fusion protein vector and its expression in E.coli[J]. Chinese Journal of Clinical Oncology and Rehabilitation, 2006, 13(3): 212-214 |
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| Authors: | MENG Yuan-guang HAN Wei-dong HUANG Ke et al |
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| Abstract: | Objective To observe and indentify the expression of human LRP16 gene in E.coli,and isolate and purify its protein product.Methods With DNA recombinant technique,the recombinant expressive plasmide pRSET-C-LRP16 was constructed and introduced into E.coli.pRSET-C-LRP16 fusion protein was expressed at the induction of IPTG.After pRSET-C-LRP16 fusion protein was purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).Results Recombinant plasmid pRSET-C-LRP16 was successfully constructed.The results of SDS-PAGE showed pRSET-C-LRP16 fusion protein was expressed in E.coli cell,and was produced at a level of 50% of the total cellar protein.Conclusions LRP16 protein could be expressed in E.coli system stably. |
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| Keywords: | LRP16 Gene fusion |
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