Abstract
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, cosl4 and cos56, which represents the HSV-1 genome was used for generation of this rHSV.Rep andcap genes of AAV-2 were inserted intoXba I site ofUL2 gene on cod, generating cos6rcΔUL2. After being digested withPac I, cos6-rcΔUL2 and the other 4 cosmids were cotransfected into BHK-21 cells. The recombinant virus HSV1-rc/ΔUL2 carryingrep andcap genes was generated due to the homologous recombination of the 5 cosmids. The results showed that the existence ofrep andcap 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.
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