Abstract: Objective To provide the material foundation for inducing pluripotent stem cells through protein transduction by constructing the recombinant expression vector TAT-OCT4 which can highly express and purify fusion protein in E.coli BL21. Methods The coding human OCT4 gene sequence was obtained by RT-PCR amplification and linked to the prokaryotic expression vector TAT-V2 for the construction of recombinant vector TAT-OCT4. E.coli BL21 was transformed into TAT-OCT4. Expression of TAT-OCT4 was induced with IPTG and identified by SDS-PAGE. The fusion protein was purified by affinity chromatography, its specificity was assayed by Western blot, and its effect on transduction of HSF cells was detected by immunofluorescence. Results The prokaryotic expression vector of TAT-OCT4 fusion protein was successfully constructed, which could highly express and purify the fusion protein as shown by Western blot. Immunofluorescence indicated that the fusion protein could be rapidly transduced into HSF cells. Conclusion TAT-OCT4 fusion protein can be safely and effectively transduced into HSF cells.