Construction of recombinant adeno-associated virus vector with human preproenkephalin gene

Objective To construct the recombinant adeno-associated virus vector with human preproenkephalin gene (rAAV-hPPE). Methods The human preproenkephalin (hPPE) gene was cloned into the adeno-associated virus (AAV) vector plasmid pSNAV which contained neo expression box. The recombinant pSNAV-hPPE was then transfected into BHK cells using lipofectamine?2000. The G418-resistant cells, BHK / SH1, were obtained. The BHK / SH1 cells were infected with HSV1-rc /△UL2 which has the function of packaging the recombinant AAV(rAAV) . After purification, the construction of rAAV-hPPE was achieved.Results The construction of pSNAV-hPPE was confirmed by digestion with restriction enzyme. Southeon bolting was used to detect the virus liters (2.5 × 1212 v.g /ml) .Conclusion This rAAV-hPPE virus vector with high liter and strong infectivity can be used in transgenic analgesic research.