Functional IBV Minigenomes Generated by Recombinant Fowl Pox Viruses for use in IBV-Targeted Recombination Studies
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Having previously demonstrated that the IBV defective (D)-RNA CD-61, can be used as an RNA vector for the expression of heterologous genes via electroporation of transcripts into infected cells (Stirrups et.al., 2000b) we wished to improve this procedure with another strategy. Hence, IBV D-RNA sequences (minigenomes) were introduced into the fowl pox virus (FPV) genome for use in a FPV/T7 system. Modified CD-61 D-RNA minigenomes were generated in situ, in a helper virus dependent manner, from cells co-infected with IBV, recombinant FPV containing CD-61 directed by the T7 promoter and rFPV expressing T7 RNA Polymerase. Efficient rescue of CD-61 D-RNAs by helper IBV was demonstrated upon serial passage in tissue culture.