Skip to main content
SpringerLink
Log in

Reverse Genetics for Porcine Reproductive and Respiratory Syndrome Virus

  • Protocol
  • First Online:
Reverse Genetics of RNA Viruses

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1602))

Abstract

Porcine reproductive and respiratory syndrome (PRRS) is a reemerging swine disease, and has become economically the most significant disease in pork production worldwide. The causative agent is PRRS virus (PRRSV), which is a member virus of the family Arteriviridae. The PRRSV genome is a single-stranded positive-sense RNA and is infectious. Two strategies in the PRRSV reverse genetics system have been employed for reconstitution of progeny virus: RNA transfection and DNA transfection. The PRRSV reverse genetics has broadly been used for studies including protein structure-function relationship, foreign gene expression, vaccine development, virulence determinants, and viral pathogenesis. Herein, we describe the modification of the pFL12 “RNA launch” reverse genetic system to the CMV promoter-driven pXJ41-FL13 “DNA launch” system. The generation of progeny PRRSV using pXJ41-FL13 is further elucidated.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Chand RJ, Trible BR, Rowland RRR (2012) Pathogenesis of porcine reproductive and respiratory syndrome virus. Curr Opin Virol 2(3):256–263. doi:10.1016/j.coviro.2012.02.002

    Article  CAS  PubMed  Google Scholar 

  2. Wensvoort G, Terpstra C, Pol JMA, Terlaak EA, Bloemraad M, Dekluyver EP, Kragten C, Vanbuiten L, Denbesten A, Wagenaar F, Broekhuijsen JM, Moonen PLJM, Zetstra T, Deboer EA, Tibben HJ, Dejong MF, Vantveld P, Groenland GJR, Vangennep JA, Voets MT, Verheijden JHM, Braamskamp J (1991) Mystery swine disease in the Netherlands—the isolation of lelystad virus. Vet Quart 13(3):121–130

    Article  CAS  Google Scholar 

  3. Allende R, Lewis TL, Lu Z, Rock DL, Kutish GF, Ali A, Doster AR, Osorio FA (1999) North American and European porcine reproductive and respiratory syndrome viruses differ in non-structural protein coding regions. J Gen Virol 80(Pt 2):307–315

    Article  CAS  PubMed  Google Scholar 

  4. Nelsen CJ, Murtaugh MP, Faaberg KS (1999) Porcine reproductive and respiratory syndrome virus comparison: divergent evolution on two continents. J Virol 73(1):270–280

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Snijder EJ, Kikkert M, Fang Y (2013) Arterivirus molecular biology and pathogenesis. J Gen Virol 94:2141–2163. doi:10.1099/Vir.0.056341-0

    Article  CAS  PubMed  Google Scholar 

  6. Fang Y, Treffers EE, Li Y, Tas A, Sun Z, van der Meer Y, de Ru AH, van Veelen PA, Atkins JF, Snijder EJ, Firth AE (2012) Efficient-2 frame shifting by mammalian ribosomes to synthesize an additional arterivirus protein. Proc Natl Acad Sci U S A 109(43):E2920–E2928. doi:10.1073/pnas.1211145109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Sawicki SG, Sawicki DL, Siddell SG (2007) A contemporary view of coronavirus transcription. J Virol 81(1):20–29. doi:10.1128/Jvi.01358-06

    Article  CAS  PubMed  Google Scholar 

  8. Han M, Yoo D (2014) Engineering the PRRS virus genome: updates and perspectives. Vet Microbiol 174(3–4):279–295. doi:10.1016/j.vetmic.2014.10.007

    Article  CAS  PubMed  Google Scholar 

  9. Meulenberg JJM, BosDeRuijter JNA, van de Graaf R, Wensvoort G, RJM M (1998) Infectious transcripts from cloned genome-length cDNA of porcine reproductive and respiratory syndrome virus. J Virol 72(1):380–387

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Lee C, Calvert JG, Welich SKW, Yoo D (2005) A DNA-launched reverse genetics system for porcine reproductive and respiratory syndrome virus reveals that homodimerization of the nucleocapsid protein is essential for virus infectivity. Virology 331(1):47–62. doi:10.1016/j.virol.2004.10.026

    Article  CAS  PubMed  Google Scholar 

  11. Fang Y, Christopher-Hennings J, Brown E, Liu H, Chen Z, Lawson SR, Breen R, Clement T, Gao X, Bao J, Knudsen D, Daly R, Nelson E (2008) Development of genetic markers in the non-structural protein 2 region of a US type 1 porcine reproductive and respiratory syndrome virus: implications for future recombinant marker vaccine development. J Gen Virol 89(Pt 12):3086–3096. doi:10.1099/vir.0.2008/003426-0

    Article  CAS  PubMed  Google Scholar 

  12. Tian DB, Zheng HH, Zhang R, Zhuang JS, Yuan SS (2011) Chimeric porcine reproductive and respiratory syndrome viruses reveal full function of genotype 1 envelope proteins in the backbone of genotype 2. Virology 412(1):1–8. doi:10.1016/j.virol.2010.12.048

    Article  CAS  PubMed  Google Scholar 

  13. Li Y, Zhou L, Zhang J, Ge X, Zhou R, Zheng H, Geng G, Guo X, Yang H (2014) Nsp9 and Nsp10 contribute to the fatal virulence of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China. Plos Pathog 10(7):e1004216. doi:10.1371/journal.ppat.1004216

    Article  PubMed  PubMed Central  Google Scholar 

  14. Ni YY, Opriessnig T, Zhou L, Cao DJ, Huang YW, Halbur PG, Meng XJ (2013) Attenuation of Porcine Reproductive and Respiratory Syndrome Virus by Molecular Breeding of Virus Envelope Genes from Genetically Divergent Strains. J Virol 87(1):304–313. doi:10.1128/Jvi.01789-12

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Zhou L, Ni YY, Pineyro P, Sanford BJ, Cossaboom CM, Dryman BA, Huang YW, Cao DJ, Meng XJ (2012) DNA shuffling of the GP3 genes of porcine reproductive and respiratory syndrome virus (PRRSV) produces a chimeric virus with an improved cross-neutralizing ability against a heterologous PRRSV strain. Virology 434(1):96–109. doi:10.1016/j.virol.2012.09.005

    Article  CAS  PubMed  Google Scholar 

  16. Zhou L, Ni YY, Pineyro P, Cossaboom CM, Subramaniam S, Sanford BJ, Dryman BA, Huang YW, Meng XJ (2013) Broadening the heterologous cross-neutralizing antibody inducing ability of porcine reproductive and respiratory syndrome virus by breeding the GP4 or M genes. PloS one 8(6):e66645. doi:10.1371/journal.pone.0066645

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ni YY, Zhao Z, Opriessnig T, Subramaniam S, Zhou L, Cao D, Cao Q, Yang H, Meng XJ (2014) Computer-aided codon-pairs deoptimization of the major envelope GP5 gene attenuates porcine reproductive and respiratory syndrome virus. Virology 450-451:132–139. doi:10.1016/j.virol.2013.12.009

    Article  CAS  PubMed  Google Scholar 

  18. Vu HL, Kwon B, de Lima M, Pattnaik AK, Osorio FA (2013) Characterization of a serologic marker candidate for development of a live-attenuated DIVA vaccine against porcine reproductive and respiratory syndrome virus. Vaccine 31(40):4330–4337. doi:10.1016/j.vaccine.2013.07.020

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Truong HM, Lu Z, Kutish GF, Galeota J, Osorio FA, Pattnaik AK (2004) A highly pathogenic porcine reproductive and respiratory syndrome virus generated from an infectious cDNA clone retains the in vivo virulence and transmissibility properties of the parental virus. Virology 325(2):308–319. doi:10.1016/j.virol.2004.04.046

    Article  CAS  PubMed  Google Scholar 

  20. Yount B, Curtis KM, Fritz EA, Hensley LE, Jahrling PB, Prentice E, Denison MR, Geisbert TW, Baric RS (2003) Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus. P Natl Acad Sci USA 100(22):12995–13000. doi:10.1073/pnas.1735582100

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This project was supported by the Agriculture and Food Research Initiative (AFRI) Competitive Grant (2013-67015-21243) of USDA NIFA (National Institute of Food and Agriculture).

Author information

Authors and Affiliations

  1. Department of Pathobiology, University of Illinois at Urbana-Champaign, Urbana, IL, 61802, USA

    Mingyuan Han, Hanzhong Ke, Yijun Du, Qingzhan Zhang & Dongwan Yoo

  2. Department of Pediatrics and Communicable Diseases, University of Michigan Medical School, Ann Arbor, MI, 48109, USA

    Mingyuan Han

  3. Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China

    Yijun Du

Authors
  1. Mingyuan Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hanzhong Ke
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yijun Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qingzhan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dongwan Yoo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dongwan Yoo .

Editor information

Editors and Affiliations

  1. Department of Population Health, Poultry Diagnostic and Research Center, College of Veterinary Medicine, University of Georgia, Athens, Georgia, USA

    Daniel R. Perez

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Han, M., Ke, H., Du, Y., Zhang, Q., Yoo, D. (2017). Reverse Genetics for Porcine Reproductive and Respiratory Syndrome Virus. In: Perez, D. (eds) Reverse Genetics of RNA Viruses. Methods in Molecular Biology, vol 1602. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6964-7_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6964-7_3

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6962-3

  • Online ISBN: 978-1-4939-6964-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation