The fused gene (TAT PTD-Ngb) which included nine amine acid transactivator of transcription (TAT) protein transduction domain (RKKRRQRRR) of HIV-1 and neuroglobin gene was amplified by RT-PCR from rat brain RNA and cloned into the expression plasmid pET28b. The recombinant plasmid pET-PN was transformed into the E.coli. BL21(DE3)plysS, which was induced with IPTG (0.4 mmol/L) to express TAT PTD-Ngb fusion protein. Ni-NTA resin was used to purify the product, which was identified by SDS-PAGE and Western blot subsequently. After being purified and desalted, the biological activity of TAT PTD-Ngb was deteced in primary cultured cortical neurons. The results showed that TAT PTD-Ngb could transduced into cortical neurons, increase cell viability under hypoxia and attenuate apoptosis induced by hypoxia. The present study provides a clue for the research of neuroglobin and seems to provide a protein therapy strategy for CNS diseases,especially in cerebrovascular diseases and neurodegenerative diseases.