A Fosmid-Based System for the Generation of Recombinant Cercopithecine Alphaherpesvirus 2 Encoding Reporter Genes

Viruses. 2019 Nov 5;11(11):1026. doi: 10.3390/v11111026.

Abstract

The transmission of Macacine alphaherpesvirus 1 (McHV-1) from macaques, the natural host, to humans causes encephalitis. In contrast, human infection with Cercopithecine alphaherpesvirus 2 (CeHV-2), a closely related alphaherpesvirus from African vervet monkeys and baboons, has not been reported and it is believed that CeHV-2 is apathogenic in humans. The reasons for the differential neurovirulence of McHV-1 and CeHV-2 have not been explored on a molecular level, in part due to the absence of systems for the production of recombinant viruses. Here, we report the generation of a fosmid-based system for rescue of recombinant CeHV-2. Moreover, we show that, in this system, recombineering can be used to equip CeHV-2 with reporter genes. The recombinant CeHV-2 viruses replicated with the same efficiency as uncloned, wt virus and allowed the identification of cell lines that are highly susceptible to CeHV-2 infection. Collectively, we report a system that allows rescue and genetic modification of CeHV-2 and likely other alphaherpesviruses. This system should aid future analysis of CeHV-2 biology.

Keywords: Cercopithecine alphaherpesvirus 2; fosmid; recombineering.

MeSH terms

  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • DNA, Viral / genetics
  • Genes, Reporter*
  • Genetic Engineering
  • Genome, Viral / genetics
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Simplexvirus / genetics*
  • Simplexvirus / physiology
  • Vero Cells
  • Viral Proteins / genetics
  • Viral Tropism
  • Virus Replication

Substances

  • DNA, Viral
  • Luminescent Proteins
  • Viral Proteins