Expression, purification and characterization of rat zinc finger protein Mipu1 in Escherichia coli
Expression, purification and characterization of rat zinc finger protein Mipu1 in Escherichia coli
复制标题
大鼠锌指蛋白 Mipu1 在大肠杆菌中的表达、纯化和表征
DOI：
10.1007/s11010-009-0083-8
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发表时间：
2009-08-01
影响因子：
4.3
通讯作者：
Xiao, Xianzhong
中科院分区：
文献类型：
--
作者：
Jiang, Lei;Zhang, Bin;Xiao, Xianzhong
新基因Mipu 1是最近在大鼠心肌缺血预处理中发现的上调基因。我们以前证明，Mipu1是一个核蛋白和转录抑制因子。本研究在大肠杆菌中表达了Mipu 1。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。获得毫克量的高度纯化的Mipu 1。纯化的蛋白质，其特征在于使用蛋白质印迹，大小排阻色谱和EMSA。用Mipu 1蛋白制备兔抗血清，通过Western blotting检测正常和应激条件下Mipu 1蛋白的表达。
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.