| 小鼠pdd87基因在大肠杆菌中的表达与纯化 |
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| 引用本文： | 段瑞峰,李刚,胡迎春,项晓琼,吴虹,吴德昌.小鼠pdd87基因在大肠杆菌中的表达与纯化[J].生物技术通讯,2004,15(4):330-332. |
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| 作者姓名： | 段瑞峰 李刚 胡迎春 项晓琼 吴虹 吴德昌 |
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| 作者单位： | 1. 军事医学科学院,放射医学研究所,北京,100850
2. 加州大学,洛杉矶分校医学院分子与医学药理系,洛杉矶,90095 |
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| 基金项目： | 国家自然科学基金(30000201)，海外青年合作研究基金(39928002) |
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| 摘 要： | 利用PCR和基因重组技术构建了三个小鼠pdd87基因的原核表达质粒：表达全长cDNA的pET-28a-pdd87质粒；表达PDD87C端404个氨基酸的pET-28a-pdd87-404质粒；表达全长cDNA的pMXB10-pdd87质粒。经IPTG诱导后三种质粒都得到表达。pET-28a-pdd87质粒和pET-28a-pdd87-404质粒表达的蛋白经His6亲和层析纯化后分别获得了带His标签的PDD87蛋白和含C端404个氨基酸的蛋白。pMXB10-pdd87质粒表达的蛋白经几丁质柱亲和层析纯化后获得了纯的PDD87蛋白。
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| 关 键 词： | 小鼠 pdd87基因 大肠杆菌 表达 纯化 亲和层析 |
| 文章编号： | 1009-0002(2004)04-0330-03 |
| 修稿时间： | 2003年12月2日 |
Expression and purification of murine pdd87 gene in E.coli |
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| DUAN Rui-feng ,LI Gang ,Matthias Groszer ,HU Ying-chun ,XIANG Xiao-qiong ,WU Hong ,Wu De-chang.Expression and purification of murine pdd87 gene in E.coli[J].Letters in Biotechnology,2004,15(4):330-332. |
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| Authors: | DUAN Rui-feng LI Gang Matthias Groszer HU Ying-chun XIANG Xiao-qiong WU Hong Wu De-chang |
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| Institution: | DUAN Rui-feng 1,LI Gang 1,Matthias Groszer 2,HU Ying-chun 1,XIANG Xiao-qiong 1,WU Hong 2,Wu De-chang 1 1.Institute of Radiation Medicine,Academy of Military Medical Sciences,Beijing100850,2.Department of Molecular and Medical Pharmacology,UCLA School of Medicine,Los Angeles,USA90095 |
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| Abstract: | Utilizing PCR and gene recombination technologies,three expression plasmids,pET-28a-pdd877,pET-28a-pdd87-404and pMXB10-pdd87,were constructed.pET-28a-pdd87and pMXB10-pdd87contain the entire murine pdd87gene ORF.pET-28a-pdd87-404contains the C-terminus404amino acids of the PDD87protein.Induced by IPTG,the three plasmids were all expressed.Purified proteins,PDD87and PDD87-404,with His 6 tag were obtained after Ni-NTA affinity chromatography.Purified PDD87protein without His 6 tag was obtained after Chitin affinity chromatography. |
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| Keywords: | pdd87gene protein express affinity chromatography protein purification |
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