Reverse Genetics for Porcine Reproductive and Respiratory Syndrome Virus

Methods Mol Biol. 2017:1602:29-46. doi: 10.1007/978-1-4939-6964-7_3.

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.

Keywords: Arterivirus; Infectious clones; Nidovirus; PRRS; Porcine reproductive and respiratory syndrome virus; Reverse genetics.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • DNA, Complementary
  • Genome, Viral
  • Plasmids / genetics
  • Porcine Reproductive and Respiratory Syndrome / virology
  • Porcine respiratory and reproductive syndrome virus / genetics*
  • Promoter Regions, Genetic
  • RNA, Viral
  • Reverse Genetics* / methods
  • Swine
  • Transfection
  • Virus Replication

Substances

  • DNA, Complementary
  • RNA, Viral