重组人核因子κB亚基p65在大肠杆菌中的可溶性表达及纯化  

Soluble Expression in E.coli and Purification of Recombinant Human Nuclear Factor κB p65 Subunit

作　　者：闫海涛[1] 马晓芸[1] 董泗建[1] 郑建全[1] 丁日高[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《生物技术通讯》2005年第6期602-604,共3页Letters in Biotechnology

摘　　要：将扩增得到的核因子(NF)κB亚基p65基因片段克隆至测序载体pGEM-T,测序验证该序列为预期目的片段后,再将该基因片段克隆至表达载体pGEX-4T2中,转化大肠杆菌BL21,IPTG诱导表达,用GST蛋白亲和柱纯化融合蛋白p65/GST,Western印迹验证该蛋白具有NF-κB抗原活性。结果表明,构建了NF-κBp65/GST融合表达载体,并表达和纯化了NF-κBp65/GST融合蛋白,为进一步研究NF-κB的功能和筛选NF-κB拮抗分子奠定了基础。An expression vector with NF-κB p65/GST was constructed and expressed in E coli BI21 induced by IPTG, and the fusion protein NF-κB p65/GST was purified by MicroSpinTM GST purification kit. The resuh of Western blot showed that this fusion protein reacted specifically with the antibody of NF-κB p65 subunit. Fusion protein NF-κB p65/ GST has been effectively expressed and purified, it will be help for the further studies of the function of NF-κB and se- lection of antibody to NF-κB.

关 键 词：核因子 p65亚基 GST 

分 类 号：Q785[生物学—分子生物学] Q786