Recovery of infectious foot-and-mouth disease virus from full-length genomic cDNA clones using an RNA polymerase I system.
The prototypic foot-and-mouth disease virus (FMDV) was shown more than a century ago to be the first filterable agent capable of causing FMD, and it has served as an important model for studying basic principles of Aphthovirus molecular biology. However, the complex structure and antigenic diversity of FMDV have posed a major obstacle to the attempts at manipulating the infectious virus by reverse genetic techniques. Here, we report the recovery of infectious FMDV from cDNAs based on an efficient in vivo RNA polymerase I (polI) transcription system. Intracellular transcription of the full-length viral genome from polI-based vectors resulted in efficient formation of infectious virus displaying a genetic marker. Compared with wild-type virus, an abundance of genomic mRNA and elevated expression levels of viral antigens were indicative of the hyperfunction throughout the life-cycle of this cDNA-derived virus at transcription, replication, and translation levels. The technology described here could be an extremely valuable molecular biology tool for studying FMDV complex infectious characteristics. It is an operating platform for studying FMDV functional genomics, molecular mechanism of pathogenicity and variation, and lays a solid foundation for the development of viral chimeras toward the prospect of a genetically engineered vaccine.
Chang
Yanyan
Y
Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou 730046, China.
Zheng
Haixue
H
Shang
Youjun
Y
Jin
Ye
Y
Wang
Guangxiang
G
Shen
Xiaoyan
X
Liu
Xiangtao
X
D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification*; D016679:Genome, Viral / Q000235:genetics*; D012318:RNA Polymerase I / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary; D012318:RNA Polymerase I