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ISSN 1672-9145                                                Acta Biochim Biophys Sin 2005, 37(3): 192–198                                                   CN 31-1940/Q

Synthesis of Reassortant Infectious Bursal Disease Virus in Chickens Injected Directly with Infectious Clones from Different Virus Strains

Long LI, Yao-Wei HUANG#, Lian-Sheng WANG, Wang-Jun WAN, and Lian YU*

 

Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Institute of Preventive Veterinary Medicine,

Zhejiang University, Hangzhou 310029, China

 

Abstract        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.

 

Key words        infectious bursal disease; infectious bursal disease virus; reverse genetics; reassortant virus; animal model

 

 

 

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Received: December 29, 2004        Accepted: January 19, 2005

This work was supported by the grants from the National High Techno­logy Research and Development Program of China (No. 2004BA757C) and the Key Project of Zhejiang Province (No. 2003C22002)

# Present address: Center for Molecular Medicine and Infectious Diseases, Virginia Polytechnic Institute and State University, USA

*Corresponding author: Tel/Fax, 86-571-86971894; E-mail, [email protected]