Synthesis of reassortant infectious bursal disease virus in chickens injected directly with infectious clones from different virus strains.
The infectious bursal disease virus (IBDV), a member of the Birnaviridae family, containing a bisegmented double-stranded RNA genome, encodes four structural viral proteins, VP1, VP2, VP3, and VP4, as well as a non-structural protein, VP5. In the present paper, the segment A from two IBDV strains, field isolate ZJ2000 and attenuated strain HZ2, were inserted into one NaeI site by site-directed silent mutagenesis and subcloned into the eukaryotic expression plasmid pCI under the control of the human cytomegalovirus (hCMV) immediate early enhancer and promoter to construct the recombinant plasmids pCI-AKZJ2000 and pCI-AKHZ2, respectively. Each of the two recombinants was combined with another recombinant pCI plasmid containing the marked segment B of strain HZ2 (pCI-mB), and injected intramuscularly into non-immunized chickens. Two chimeric IBDV strains were recovered from the chickens. Two out of eight chickens in each of two groups showed the bursal histopathological change. The reassortant virus derived from pCI-AKZJ2000/pCI-mB can infect chicken embryos and shows relatively low virulence. We have developed a novel virus reverse genetic approach for the study of IBDV. The results also form the basis for investigating the role of VP1 in viral replication and pathogenecity.
Li
Long
L
Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, China.
Huang
Yao-Wei
YW
Wang
Lian-Sheng
LS
Wan
Wang-Jun
WJ
Yu
Lian
L
D000818:Animals; D018175:Birnaviridae Infections / Q000821:virology*; D002642:Chick Embryo; D002645:Chickens / Q000821:virology*; D019976:Cloning, Organism / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D007243:Infectious bursal disease virus / Q000235:genetics* / Q000472:pathogenicity*; D007267:Injections; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D009154:Mutation / Q000235:genetics; D016865:Reassortant Viruses / Q000235:genetics* / Q000472:pathogenicity*; D013045:Species Specificity