小鼠pdd87基因在大肠杆菌中的表达与纯化  

Expression and purification of murine pdd87gene in E.coli

作　　者：段瑞峰[1] 李刚[1] Matthias Groszer 胡迎春[1] 项晓琼[1] 吴虹[2] 吴德昌[1] 

机构地区：[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

出　　处：《生物技术通讯》2004年第4期330-332,共3页Letters in Biotechnology

基　　金：国家自然科学基金(30000201);海外青年合作研究基金(39928002)

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。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.

关 键 词：小鼠 pdd87基因 大肠杆菌 表达 纯化 亲和层析 

分 类 号：Q786[生物学—分子生物学]