Jiang Lei, Zhang Bin, Wang Guiliang, Wang Kangkai, Xiao Xianzhong
Department of Pathophysiology, Central South University, Changsha, Hunan, People's Republic of China.
Mol Cell Biochem. 2009 Aug;328(1-2):137-44. doi: 10.1007/s11010-009-0083-8. Epub 2009 Apr 1.
The novel gene Mipu1 was recently identified in rat due to its up-regulation in response to myocardial ischemia preconditioning. We previously demonstrated that Mipu1 was a nuclear protein and a transcriptional repressor. In this study, Mipu1 was expressed in E. coli and purified using a recombinant expression system and a purification protocol. Milligram quantities of highly purified Mipu1 were obtained. The purified protein was characterized using western blotting, size exclusion chromatography and EMSA. The Mipu1 protein was also used to generate antiserum in rabbits, which was used to detect the expression of Mipu1 protein under normal and stress conditions, by western blotting.
新型基因Mipu1最近在大鼠中被发现，因为它在心肌缺血预处理反应中上调。我们之前证明Mipu1是一种核蛋白和转录抑制因子。在本研究中，Mipu1在大肠杆菌中表达，并使用重组表达系统和纯化方案进行纯化。获得了毫克量的高度纯化的Mipu1。使用蛋白质免疫印迹、尺寸排阻色谱和电泳迁移率变动分析对纯化的蛋白质进行表征。Mipu1蛋白还用于在兔中产生抗血清，该抗血清通过蛋白质免疫印迹用于检测正常和应激条件下Mipu1蛋白的表达。
