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Engineering infectious foot-and-mouth disease virus in vivo from a full-length genomic cDNA clone of the A/AKT/58 strain

  • In Memoriam: Professor Ray Wu
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Abstract

Two full-length genomic cDNA clones, pTA/FMDV and pCA/FMDV, were constructed that contained three point-mutants [A174G and A308G (not present in pTA/FMDV); T1029G] in the genome compared with the wild type A/AKT/58 strain of foot-and-mouth disease virus. These two viruses were rescued by co-transfection of pCA/FMDV with pCT7RNAP, which can express T7 RNA polymerase in BHK-21 cell-lines, or by transfection of the in vitro transcribed RNA. Their biological properties were analyzed for their antigenicity, virulence in suckling-mice (LD50) and growth kinetics in BHK-21 cells. The in vivo rescued viruses showed high pathogenicity for 3-day-old unweaned mice (LD50=10−7.5). However, the in vitro transcribed RNA derived from pTA/FMDV had lower pathogenicity for suckling-mice (LD50=10−6), and the in vivo transcribed RNA recovered from pCA/FMDV co-transfected with pCT7RNAP showed no significant differences from the wild type virus. These data showed that recovery of the infectious foot-and-mouth disease virus directly from the use of in vivo techniques was better than from in vitro methods. Furthermore, the reverse genetic procedure technique was simplified to a faster one-step procedure based on co-transfection with pCT7RNAP. These results suggest that in vivo RNA transcripts may be more valuable for engineering recombinant foot-and-mouth disease virus than in vitro RNA transcripts, and may contribute to further understanding of the biological properties, such as replication, maturation and quasispecies, of the foot-and-mouth disease virus.

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Authors and Affiliations

  1. Key Laboratory of Animal Virology of Ministry of Agriculture, National Foot-and-Mouth Disease Reference Laboratory of China, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China

    XingWen Bai, PingHua Li, YiMei Cao, Dong Li, ZengJun Lu, JianHong Guo, DeHui Sun, Pu Sun, XiangTao Liu, JianXun Luo & ZaiXin Liu

  2. Veterinary Research Institute, Guangdong Agricultural Academy of Sciences, Guangzhou, 510640, China

    HaiXue Zheng

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Supported by the National Key Basic Research Program of China (Grant No. 2005CB523201) and National High-Tech Research and Development Program of China (Grant No. 2006BAD06A03)

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Bai, X., Li, P., Cao, Y. et al. Engineering infectious foot-and-mouth disease virus in vivo from a full-length genomic cDNA clone of the A/AKT/58 strain. SCI CHINA SER C 52, 155–162 (2009). https://doi.org/10.1007/s11427-009-0007-6

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