The generation of a recombinant HSV (rHSV) that can provide packaging function for rAAV production is described. A set of cosmids including cos48, cos28, cos6, cos14 and cos56, which represents the HSV-1 genome was used for generation of this rHSV. Rep and cap genes of AAV-2 were inserted into Xba Ⅰ site of UL2 gene on cos6, generating cos6-rcΔUL2. After being digested with Pac Ⅰ , cos6-rcΔUL2 and the other 4 cosmids were cotrans-fected into BHK-21 cells. The recombinant virus HSV1-rc/ΔUL2 carrying rep and cap genes was generated due to the homologous recombination of the 5 cosmids. The results showed that the existence of rep and cap genes on this rHSV was stable from passage to passage and the rHSV could support the packaging of rAAV either in cells transiently transfected with AAV vector or in stable cell line harboring AAV vector. Further modification of this rHSV and optimization of conditions involved in rAAV preparation may lead to a large-scale production of rAAV in the near future. The generation of a recombinant HSV (rHSV) that can provide packaging function for rAAV production is described. A set of cosmids including cos48, cos28, cos6, cos14 and cos56, which represents the HSV-1 genome was used for generation of this rHSV. Rep and cap genes of AAV-2 were inserted into Xba I site of UL2 gene on cos6, generating cos6-rcΔUL2. After being digested with Pac I, cos6-rcΔUL2 and the other 4 cosmids were cotransfected into BHK-21 cells. The recombinant of HSV1-rc / ΔUL2 carrying rep and cap genes was generated due to the homologous recombination of the 5 cosmids. The results showed that the existence of rep and cap genes on this rHSV was stable from passage to passage and the rHSV could support the packaging of rAAV either in cells transiently transfected with AAV vector or in stable cell line harboring AAV vector. Further modification of this rHSV and optimization of conditions involved in rAAV preparation may lead to a large-scale production of rAAV in the near future