Abstract
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.
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References
Hiscox, J. A.., Mawditt, K. L., Cavanagh, D. & Britton, P. (1995). Investigation of the control of Coronavirus subgenomic mRNA transcription by using T7-generated negative sense RNA transcripts. J. Virol. 69, 6219–6227.
Makino, S. & Lai, M. M. C. (1989). High-frequency leader sequence switching during Coronavirus defective interfering RNA replication. J. Virol. 63, 5285–5292.
Penzes, Z., Tibbies, K., Shaw, K., Britton, P., Brown, T. D. K. & Cavanagh, D. (1994). Characterisation of a replicating and passaged defective RNA of avian Coronavirus infectious bronchitis virus. Virology. 203, 286–293.
Stirrups, K., Shaw, K., Evans, S., Dalton, K., Cavanagh, D. & Britton, P. (2000a). Leader switching occurs during the rescue of defective RNAs by heterologous strains of the Coronavirus infectious bronchitis virus (IBV). J. Gen. Virol. 81, 791–801.
Stirrups, K., Shaw, K., Evans, S., Dalton, K., Casais, R., Cavanagh, D. & Britton, P. (2000b). Expression of reporter genes from the defective RNA CD-61 of the Coronavirus infectious bronchitis virus. J. Gen. Virol. 81, 1687–1698.
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Evans, S., Cavanagh, D., Britton, P. (2001). Functional IBV Minigenomes Generated by Recombinant Fowl Pox Viruses for use in IBV-Targeted Recombination Studies. In: Lavi, E., Weiss, S.R., Hingley, S.T. (eds) The Nidoviruses. Advances in Experimental Medicine and Biology, vol 494. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-1325-4_78
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DOI: https://doi.org/10.1007/978-1-4615-1325-4_78
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