Expression of hNeuritin protein in a baculovirus expression system and the analysis of its activity

Neuritin plays an important role in the development and regeneration of the nervous system, and shows good prospects in the treatment and protection of the nervous system. To characterize neuritin function, we constructed a baculovirus expression system of neuritin, and identified the biological activity of the neuritin protein. The results and showed that the expression product could promote the neurite growth of dorsal root ganglion in chicken embryos. The neuritin open reading frame was amplified and cloned into the plasmid pFastBac™HTA. The pFastBac™HTA-neuritin was confirmed to be correct by PCR and DNA sequencing, and then transformed into Escherichia coli DH10Bac. The high purity recombinant Bacmid-neuritin (shuttle vectors) was obtained from DH10Bac through screening and identification. Recombinant virus, including the neuritin gene (virus-neuritin), was produced by transfection of SF9 cells using the bacmid-neuritin, and then amplified repeatedly to express the neuritin fusion protein. Finally, we identified the fusion protein with SDS-PAGE and western blotting, and optimized the best expression time of the neuritin fusion protein. We also analyzed the activity of the expressed protein by dorsal root ganglion from chicken embryos.