[Attenuation of a genotype VIId Newcastle disease virus ZJI strain of goose origin by reverse genetics].
Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain isolated from an outbreak in the goose, seven pairs of primers were designed to amplify cDNA fragment for constructing the plasmid pNDV/ZJI, which contained the full-length cDNA of NDV ZJI strain. The pNDV/ZJI with three helper plasmids, pCI-NP, pCI-P and pCI-L, were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase. After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock, infectious NDV ZJI strain was successfully rescued. The recombinant plasmid pNDV/ZJIFM was generated by converting the multi-basic amino acid sequence of the F0 protein cleavage region in pNDV/ZJI to the non-basic amino acid sequence characteristic of avirulent NDV strain. After cotransfection of the resultant plasmid and the three helper plasmids into BSR-T7/5 cells, the recombinant NDV, NDV/ZJIFM, was generated. The mean death time (MDT) of NDV/ZJIFM was more than 120h and the intrancerebral pathogenicity index (ICPI) was 0.16, indicating that the rescued virus was highly attenuated. This attenuated genotype VIId NDV of goose origin could be a desirable vaccine in controlling the current epidemic of ND.
Hu
Shun-lin
SL
Animal Infectious Disease Laboratory, School of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Sun
Qing
Q
Wu
Yan-tao
YT
Zhang
Yan-mei
YM
Liu
Xiu-fan
XF
D000818:Animals; D002642:Chick Embryo; D005777:Geese / Q000821:virology*; D005838:Genotype; D009522:Newcastle disease virus / Q000235:genetics* / Q000472:pathogenicity; D010957:Plasmids; D011995:Recombination, Genetic; D014613:Vaccines, Attenuated / Q000276:immunology; D014760:Viral Fusion Proteins / Q000235:genetics; D014765:Viral Vaccines / Q000276:immunology
D014613:Vaccines, Attenuated; D014760:Viral Fusion Proteins; D014765:Viral Vaccines