Development of a reverse genetics system based on RNA polymerase II for Newcastle disease virus genotype VII.
Newcastle disease virus (NDV) has only a single serotype but diversified genotypes. Genotype VII strains are the prevalent currently circulating genotype worldwide, and in particular, these strains cause outbreaks in waterfowl. In this study, a reverse genetics system for highly virulent NDV isolated from goose flocks was developed independent of conventional T7 RNA polymerase. Infectious virus was successfully generated by an RNA polymerase II promoter to drive transcription of the full-length virus antigenome. A green fluorescent protein (GFP)-expressing virus was generated by inserting an additional transcription cassette coding for the enhanced GFP between the P and M genes of the genome. The expression of GFP was confirmed by western blotting and fluorescence microscopy. The replication kinetics and pathogenicity of the recombinant viruses are indistinguishable from the parental wild-type virus. This reverse genetics system will provide a powerful tool for the analysis of goose-origin NDV dissemination and pathogenesis, as well as preparation for genotype-matched NDV attenuated vaccines.
Laboratory of Infectious Diseases, College of Veterinary Medicine, Jilin University, Changchun, 130062, China.
Jianzhong
J
Wang
Chunfeng
C
Wang
Na
N
Feng
Hualei
H
Wang
Xuexing
X
Zheng
Songtao
S
Yang
Yuwei
Y
Gao
Xianzhu
X
Xia
Renfu
R
Yin
Xiufan
X
Liu
Shunlin
S
Hu
Chan
C
Ding
Shengqing
S
Yu
Yanlong
Y
Cong
Zhuang
Z
Ding
D000818:Animals; D019677:Artificial Gene Fusion; D015153:Blotting, Western; D005777:Geese / Q000821:virology; D017930:Genes, Reporter; D016679:Genome, Viral; D005838:Genotype; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008856:Microscopy, Fluorescence; D009522:Newcastle disease virus / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity / Q000502:physiology; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D059386:Reverse Genetics / Q000379:methods*; D014158:Transcription, Genetic; D014773:Virology / Q000379:methods*; D014779:Virus Replication
D049452:Green Fluorescent Proteins; D012319:RNA Polymerase II