An improved method for infectious bursal disease virus rescue using RNA polymerase II system.
Reverse genetics system is an excellent platform to research the construction and function of viruses. Genome modification, such as gene recombination, mosaicism, and mutation may interfere with replication, assembly and release of viruses. An efficient, convenient and economical method of virus rescue is undoubtedly required for elevating the efficiency of rescuing crippled virus. In this study, we developed a method to rescue infectious bursal disease virus (IBDV) using RNA polymerase II. The genome of IBDV Gt strain, flanked by hammerhead ribozyme and hepatitis delta ribozyme sequences, were cloned downstream of the cytomegalovirus enhancer and the beta chicken actin promoter of the vector pCAGGS. Through direct transfection in various cell lines, IBDV could be rescued efficiently. The RNA polymerase II-based reverse genetics system is efficient, stable, convenient, and fit to various cells. The system not only provides the basis of the gene function research of IBDV, but is also useful for reverse genetics research of other birnaviridae viruses.
Qi
Xiaole
X
Division of Avian Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
Gao
Yulong
Y
Gao
Honglei
H
Deng
Xiaoyun
X
Bu
Zhigao
Z
Wang
Xiaoyan
X
Fu
Chaoyang
C
Wang
Xiaomei
X
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary; D016679:Genome, Viral; D007243:Infectious bursal disease virus / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D010957:Plasmids; D012319:RNA Polymerase II / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*; D014709:Vero Cells; D014773:Virology / Q000379:methods
D018076:DNA, Complementary; D012319:RNA Polymerase II