The deletion of an extra six nucleotides in the 5' -untranslated region of the nucleoprotein gene of Newcastle disease virus NA-1 decreases virulence

BMC Vet Res. 2014 Dec 21:10:964. doi: 10.1186/s12917-014-0305-5.

Abstract

Background: The virulent Newcastle disease virus (NDV) strain NA-1 (genotype VII) was isolated from an epizootic involving a flock of geese in Jilin Province, Northeast China, in 1999. Compared with the classical NDV strains, which have a genome size of 15,186 bp, the more recently isolated NDV strains, including that involved in the goose outbreak, have an extra six nucleotides in the 5'-untranslated region (UTR) of the nucleoprotein (NP) gene. This extra sequence, TCCCAC, is highly conserved and has been found in multiple NDV strains, including ZJ-1, WF00G, JSG0210, and NA-1. In the current study, an infectious clone from strain NA-1 was isolated and designated rNA-1. Subsequently, strain rNA-1 was mutated to delete the six-nucleotide insertion, producing strain rNA-1(-). Virulence of the recombinant virus was then assayed in chickens and geese.

Results: The recombinant virus rNA-1(-) showed similar biological characteristics to the parental NA-1 strain in DF-1 chicken fibroblast cells. However, the virulence of rNA-1(-) in 9-day-old embryonated chicken eggs and 1-day-old specific pathogen-free (SPF) chickens was decreased compared with the rNA-1 control. Furthermore, the virulence of the recombinant strain was slightly decreased in 1-day-old SPF chickens when compared with that in 1-day-old geese.

Conclusion: Following deletion of six nucleotides in the 5'-UTR of the NP gene of NDV strain NA-1, the virulence of the rNA-1(-) recombinant strain was decreased in both chickens and geese. However, rNA-1(-) was more virulent in chickens than geese, as seen by the higher intracerebral pathogenicity index value.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Untranslated Regions / genetics
  • Animals
  • Blotting, Western / veterinary
  • Chickens / virology
  • Cloning, Molecular
  • Geese / virology
  • Gene Deletion*
  • Newcastle Disease / virology
  • Newcastle disease virus / genetics*
  • Newcastle disease virus / pathogenicity
  • Nucleoproteins / genetics*
  • Real-Time Polymerase Chain Reaction / veterinary
  • Virulence / genetics

Substances

  • 5' Untranslated Regions
  • Nucleoproteins