Expression, purification and characterization of rat zinc finger protein Mipu1 in Escherichia coli.

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.