Expression and Purification of Cell Penetrating Peptide TAT and Mouse Stem Cell Transcriptional Factor Oct4 Recombinant Protein

Abstract:

In this study,TAT-Oct4 target gene was inserted into pET28a expression vector to construct pET28a-TAT-Oct4 recombinant expression vector. TAT-Oct4 fusion protein was mainly expressed with insoluble inclusion bodies in E.coli. The purity of expression product was more than90% after purification. And the average refolding field is 8.8% using urea gradient dialysis.Immunocytochemistry analysis showed that TAT-Oct4 recombinant protein could penetrate nearly100% cells membrane and was mainly concentrated in nucleus. The method of generating active TAT-Oct4 recombinant protein was established in this article. It provided not only effective study materials for cell reprogramming but also practical method for other reprogramming factors design.