Prokaryotic expression of buffalo iPSCs transcription factor Sox2 and assist protein HA2-TAT

To obtain Sox2 protein which could penetrate cell membrane,buffalo iPSCs(induced pluripotent stem cells) transcription factor Sox2 and cell-penetrating peptide HIV TAT were fusingly expressed.First,HA2-TAT fragment was synthesised to construct a basic expressing plasmid,pET-HA2-TAT.Then buffalo Sox2 gene was subcloned to pET-HA2-TAT,forming a recombinant plasmid pET-NLS-Sox2-TAT.It was transformed into E.coli BL21(DE3) and the fusion protein was expressed with induction of IPTG.SDS-PAGE analysis and Western blot was performed to detect the fusion protein which was isolated and purified by Ni2+ protein purificating column.The results showed that the fusion protein HA2-TAT(24 400)and NLS-Sox2-TAT(57 700) could high efficiently expressed.When the imidazole at the density of 365.30 mmol/L,fusion protein NLS-Sox2-TAT emerges eluting peak.The fusion protein could be detected with biology antigenicity by Western blot.