Establishment of Reverse Genetics System for NDV Isolated from Goose

Abstract:

Eight fragments were amplified and cloned into pCR2.1 vector with the designed primers. The fragments, amplified with primer Ⅰ to Ⅶ, were subcloned into transcription vector to construct the plasmid pNDVZJI which contained the full-length cDNA of NDV ZJI strain. The eukaryotic expression vector pCI-L was constructed by subcloning the fragments, amplified with the primer Ⅴ, Ⅵ and Ⅷ, into the expression vector pCI-neo. The full-length cDNA clone, pNDVZJI, with three helper plasmids, pCI-NP、pCI-P and pCI-L, were cotranfected into BSR-T7/5 cell expressing T7 RNA polymerase. After inoculation of transfected cell culture into embryonated chicken eggs from specific pathogen free(SPF) flock, The NDV of ZJI strain was rescued successfully, which laid a good foundation for the further related research.

Key words:Newcastle disease virus, cDNA, Transfection, Rescue