Use of Defective RNAs Containing Reporter Genes to Investigate Targeted Recombination for Avian Infectious Bronchitis Virus
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At present there is no infectious cDNA of IBV and the robustness of Coronavirus cDNA reverse genetic systems remains undetermined. We have been investigating an alternate strategy for reverse genetic manipulation of IBV using defective RNAs (D-RNAs) to exchange genetic material with the IBV genome, based on efficient recombination systems for other coronaviruses. It has been previously shown that two strains of IBV can recombine in ovo (Kottier et al., 1995) though recombination has not yet been demonstrated in cell culture. IBV D-RNA CD-91 (Pénzes et al., 1994) consisting of regions from the 5’-end, open reading frame 1b and the 3’-end including most of the N gene was isolated and cloned after high multiplicity passage of IBV Beaudette. Internal sequences were deleted from CD-91 to generate CD-61 (Pénzes et al., 1996). Both D-RNAs contain all the necessary signals for replication and packaging (rescue) by helper IBV.
KeywordsGene Cassette Plaque Assay Infectious Bronchitis Virus EGFP Gene Reverse Genetic System
- Stirrups, K. Shaw, K., Evans, S., Dalton, K., Casais, R., Cavanagh, D. & Britton, P. Expression of reporter genes from the defective RNA CD-61 of the Coronavirus infectious bronchitis virus. J. Gen. Virol. 81, 1687–1698.Google Scholar