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Development of a novel single-step reverse genetics system for the generation of classical swine fever virus

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Abstract

We describe an alternative reverse genetics system for generating classical swine fever virus (CSFV) based on swine RNA polymerase I promoter (pSPI)-mediated vRNA transcription. The recombinant plasmid pSPTI/SM harboring a full-length CSFV Shimen strain cDNA, flanked by a swine RNA polymerase I (pol I) promoter sequence at the 5′ end and a murine pol I terminator sequence at the 3′ end, was constructed. When the plasmid pSPTI/SM was introduced into PK-15 cells by transfection, an infectious CSFV with termini identical to those of the parental virus was generated directly. CSFV rescued from this reverse genetics system exhibited similar growth kinetics and plaque formation compared with the parental CSFV. When the novel reverse genetics system was used to generate the CSFV vaccine C-strain, infectious virus was detected in the supernatant of PK-15 cells transfected with the recombinant plasmid pSPTI/C. This novel reverse genetics system is a simple and efficient tool for the investigation of the structure and function of the viral genome, for molecular pathogenicity studies, and for the development of genetically engineered vaccines for CSFV.

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Acknowledgments

We were grateful to Dr. N. Ruggli and Dr. J.-D. Tratschin for kindly providing the plasmid pACNR1180. This study was funded by National Natural Science Foundation of China (31070134 and 31272585).

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Authors and Affiliations

  1. State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, China

    Ling Li, Huining Pang, Rui Wu, Yanwen Zhang, Yiluo Tan & Zishu Pan

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  1. Ling Li
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  2. Huining Pang
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  3. Rui Wu
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  4. Yanwen Zhang
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Correspondence to Zishu Pan.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Li, L., Pang, H., Wu, R. et al. Development of a novel single-step reverse genetics system for the generation of classical swine fever virus. Arch Virol 161, 1831–1838 (2016). https://doi.org/10.1007/s00705-016-2851-6

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  • DOI: https://doi.org/10.1007/s00705-016-2851-6

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