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Acta Biochimica et Biophysica Sinica Advance Access published online on November 12, 2009
Acta Biochimica et Biophysica Sinica, doi:10.1093/abbs/gmp093
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© The Author 2009. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

Recovery of infectious foot-and-mouth disease virus from full-length genomic cDNA clones using an RNA polymerase I system

Yanyan Chang, Haixue Zheng, Youjun Shang, Ye Jin, Guangxiang Wang, Xiaoyan Shen and Xiangtao Liu*

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

* Correspondence address. Tel: +86-931-8342710; Fax: +86-931-8342052; E-mail: hnxiangtao{at}hotmail.com


  Abstract

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.

Keywords    foot-and-mouth disease virus; RNA polI-based system; infectious cDNA clone

Received: May 11, 2009; Accepted: August 10, 2009
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