Construction of the vector of human nerve growth factor recombinant adeno-associated virus / 西安交通大学学报(医学版)

ABSTRACT

Objective To investigate the treatment of spinal cord injury (SCI) by recombinant adeno-associated virus (AAV) transducing the hNGF gene, and construct and produce the vector of hNGF recombinant AAV. Methods The resulting gene of hNGF was inserted into the KpnⅠ-BamHⅠ site of vector plasmid pSSHG-Neo to construct the vector of hNGF recombinant AAV. The recombinant AAV viral stock was packaged. Renal embryo 293 cell was co-transfected with the rAAV vector of plasmid pSSHG/hNGF, packaging plasmid pAAV/Ad and helper adenovirus pasmid pFG140 instead of adenovirus by calcium phosphate precipitation. Results The recombinant viral stock vector of plasmid pSSHG/hNGF was constructed successfully. The results of dot blot showed that we had obtained the rAAV stocks of high titre 1.46?10 12 PFU?mL -1. Conclusion We prepared the viral stock of rAAV-hNGF that can serve as the experimental study of gene therapy of SCI.