Establishment of infectious bursal disease virus(IBDV) reverse genetics system using T7 RNA polymerase
基于T7 RNA聚合酶的传染性法氏囊病病毒反向遗传系统的建立

In order to establish the reverse genetics of IBDV,the sequences of T7 promoter and ribozyme HDV were introduced into the 5' and 3' terminals of segments A and B of IBDV genome using PCR.Then the segments,which containing promoter and ribozyme HDV sequences,were cloned into PT-Pluc vector and named as PT-mA and PT-mB, respectively.Both PT-mA and PT-mB were co-transfected into vero cells which were infected by recombinant vaccinia virus vTF7-3(vTF7-3 can express the T7 RNA polymerase in vero cell) for 1 hour.The surveying of cytopathic effect(CPE),indirect immunofluorescence test and molecular marker detection confirmed the succeeding of the rescuing of IBDV.