[Cloning of infectious bursal disease virus (Gt strain) lack of VP5 gene].
Infectious bursal disease virus (IBDV) belongs to genus Avibirnavirus of the family Birnaviridae. The genome of IBDV consists of two segments of double-strand RNA, which encode four structural protein VP1-VP4 and one non-structural protein VP5.


OBJECTIVE
To study the function of VP5 of IBDV, the recombinant virus, lack of VP5 gene, was constructed and rescused by reverse genetic technique.


METHODS
We deleted the VP5 gene (on segment A) of IBDV Gt strain by silence the start codon (ATG-ATC) using site-directed mutagenesis. The full length cDNA of segment A was flanked by hammerhead ribozyme and hepatitis delta virus ribozyme, which was introduced into an eukaryotic expression vector PCAGGS, under the strong chicken beta actin promoter. The recombinant plasmid was named as pCAGGmGtA deltaVP5HRT. Co-transfection was carried with PCAGGmGtAdVP5HRT and PCAGGmGtBHRT in DF-I cells.


RESULTS
Recombinant virus was successfully rescused, which was verified by RT-PCR and indirect immnuofluorescence assay. The rescusd virus could be a very helpful platform for further study of VP5 biological function.
Qin
Liting
L
Division of Avian Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. qinliting@yahoo.com.cn
Qi
Xiaole
X
Gao
Yulong
Y
Gao
Honglei
H
Bu
Zhigao
Z
Wang
Xiaomei
X
D000818:Animals; D002645:Chickens; D002522:Chlorocebus aethiops; D007243:Infectious bursal disease virus / Q000235:genetics* / Q000502:physiology; D016297:Mutagenesis, Site-Directed; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017361:Viral Nonstructural Proteins / Q000172:deficiency* / Q000235:genetics; D014779:Virus Replication / Q000235:genetics*
C518635:VP5 protein, infectious bursal disease virus; D017361:Viral Nonstructural Proteins