Sequence analysis of the large (L) polymerase gene and trailer of the peste des petits ruminants virus vaccine strain Nigeria 75/1: expression and use of the L protein in reverse genetics

Virus Res. 2009 Oct;145(1):9-17. doi: 10.1016/j.virusres.2009.06.002. Epub 2009 Jun 18.

Abstract

The large (L) polymerase gene and the 5'-terminal UTR of the genome of peste des petits ruminants virus (PPRV), vaccine strain Nigeria 75/1, were cloned and sequenced. The L protein was also expressed in eukaryotic cells and its polymerase activity was quantitatively measured in a PPR reverse genetics assay using a reporter minigenome. Comparative sequence analysis of this functional L gene with corresponding genes of other morbilliviruses showed a degree of conservation exceeding 70%. The multiple sequence alignment and the phylogenetic study of L gene discriminated the morbilliviruses in 6 clusters, which are more closely related to Tupaia and Henipaviruses than to other paramyxoviruses. Important protein domains and functional motifs of the L polymerase of the PPRV Nigeria 75/1 vaccine were also identified by using different bioinformatics tools.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chlorocebus aethiops
  • DNA-Directed RNA Polymerases / biosynthesis
  • DNA-Directed RNA Polymerases / genetics
  • Genes, Viral*
  • Genome, Viral
  • Molecular Sequence Data
  • Peste-des-Petits-Ruminants / metabolism
  • Peste-des-Petits-Ruminants / virology*
  • Peste-des-petits-ruminants virus / genetics*
  • Peste-des-petits-ruminants virus / metabolism
  • Phylogeny
  • RNA, Viral / analysis
  • RNA, Viral / genetics
  • Sequence Alignment
  • Sequence Analysis, RNA
  • Vero Cells
  • Viral Proteins / genetics*

Substances

  • RNA, Viral
  • Viral Proteins
  • DNA-Directed RNA Polymerases