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| 人催乳素受体胞外区蛋白的原核表达及纯化 | |
| 引用本文： | 刘雪瑶,鲁雅洁,屈芫,姚俊,魏钦俊,曹新,冯振卿. 人催乳素受体胞外区蛋白的原核表达及纯化[J]. 生物技术通讯, 2010, 21(1): 7-11. DOI: 10.3969/j.issn.1009-0002.2010.01.002 |
| 作者姓名： | 刘雪瑶 鲁雅洁 屈芫 姚俊 魏钦俊 曹新 冯振卿 |
| 作者单位： | 1. 南京医科大学,基础医学院,江苏,南京,210029 2. 南京医科大学,基础医学院,江苏,南京,210029;南京医科大学,卫生部抗体技术重点实验室,江苏,南京,210029 3. 南京医科大学,卫生部抗体技术重点实验室,江苏,南京,210029 |
| 基金项目： | 江苏省自然科学基金，中国博士后科学基金，江苏省博士后科研资助计划，南京医科大学科技发展基金 |
| 摘 要： | 目的:建立人催乳素受体的原核表达系统,并在大肠杆菌中获得表达。方法:由RT-PCR获得人催乳素受体(human prolactin receptor,hPRLR)胞外区氨基酸的编码序列,扩增并通过酶切位点修饰后克隆至pMD18-T载体,经测序正确后,切下编码序列连接到重组表达载体pGEX-4T-2中,转化大肠杆菌Rosetta(DE3),用IPTG诱导重组工程菌表达,使用谷胱甘肽偶联的GSTrapFF柱亲和层析纯化重组蛋白。结果:重组菌株可以表达GST-hPRLR融合蛋白,用免疫印迹反应鉴定纯化的融合蛋白,在相对分子质量为37.6×103处有一条带。结论:利用大肠杆菌表达系统获得了较高纯度的GST-hPRLR融合蛋白,为进一步研究催乳素受体的功能和制备特异性的抗体奠定了基础。 |
| 关 键 词： | 人催乳素受体 融合蛋白 原核表达 纯化 |
Prokaryotic Expression and Purification of the Extraceliuar Domain of Human Prolactin Receptor |
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| LIU Xue-Yao,LU Ya-Jie,QU Yuan,YAO Jun,WEI Qin-Jun,CAO Xin,FENG Zhen-Qing. Prokaryotic Expression and Purification of the Extraceliuar Domain of Human Prolactin Receptor[J]. Letters in Biotechnology, 2010, 21(1): 7-11. DOI: 10.3969/j.issn.1009-0002.2010.01.002 | |
| Authors: | LIU Xue-Yao LU Ya-Jie QU Yuan YAO Jun WEI Qin-Jun CAO Xin FENG Zhen-Qing |
| Affiliation: | LIU Xue-Yao1,LU Ya-Jie1,QU Yuan1,YAO Jun1,WEI Qin-Jun1,CAO Xin1,2,FENG Zhen-Qing2 1.School of Basic Medical Sciences,Nanjing Medical University,Nanjing 210029,2.Key Laboratory of Antibody Technique of Ministry of Health,China |
| Abstract: | Objective:To construct prokaryotic expression vector carrying DNA fragment encoding human prolactin receptor(hPRLR)and to express hPRLR in E.coli.Methods:By RT-PCR,the coding sequence of the extracellular domain of hPRLR fragment was obtained and cloned into pMD18-T.DNA fragment sequenced correctly was ligated into pGEX-4T-2.E.coli Rosetta(DE3)strain was transformed by recommended pGEX-4T-2 and induced by IPTG to express fusion protein GST-hPRLR.Results:Expressing plasmid pGEX-4T-2-hPRLR was obtained.The pu... |
| Keywords: | human prolactin receptor fusion protein prokaryotic expression purification |
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