Virus Research 136 (2008) 192–197 Contents lists available at ScienceDirect Virus Research journal homepage: www.elsevier.com/locate/virusres Short communication Full genome sequences of two virulent strains of peste-des-petits ruminants virus, the C ˆ ote d’Ivoire 1989 and Nigeria 1976 strains Louisa S. Chard a,∗ , Dalan S. Bailey b , Pradyot Dash c , Ashley C. Banyard a , Thomas Barrett a a Pirbright Laboratory, Institute for Animal Health, Ash Road, Woking, Surrey GU24 0NF, United Kingdom b Department of Virology, Faculty of Medicine, Imperial College London, St Mary’s Campus, Norfolk Place, London W2 IPG, United Kingdom c Department of Immunology, Doherty Laboratory, St. Jude Children’s Research Hospital, 332 N Lauderdale, Mail Stop 351, Memphis, TN, 38105, USA article info Article history: Received 7 December 2007 Received in revised form 7 April 2008 Accepted 22 April 2008 Available online 9 June 2008 Keywords: Morbillivirus Pestes-des-petits ruminants Genome Promoters Proteins abstract Peste-des-petits ruminants virus (PPRV) causes acute febrile illness in both farmed and wild small rumi- nants, with associated mortality rates of 50–80%. PPRV is a member of the Morbillivirus genus within the Paramyxovirus family and although there are many full length genome sequences available for members of this family, their availability for PPRV in particular is limited. We have determined the full length sequences representing two virulent strains of PPRV, the Cˆ ote d’Ivoire 1989 (CI/89) and Nigeria 1976 (Ng76/1) strains. We present an alignment of the promoter regions of these viruses with other available PPRV promoter sequences and have identified domains in PPRV proteins believed to be critical for paramyxovirus pro- moter attenuation. We have also analysed the proteins of these viruses, comparing them to other available PPRV protein sequences and identified motifs that were previously recognised as being required for the function of other paramyxovirus proteins. © 2008 Elsevier B.V. All rights reserved. Peste-des-petits ruminants (PPR) is a highly contagious dis- ease of small ruminants that is widespread throughout some of the poorest developing regions of the world. It was first described in west-Africa in the 1940s (Gargadennec and Lalanne, 1942) and continues to circulate in sub-saharan Africa, the middle-east and southern Asia. More recently an outbreak has been reported in the Tibetan region of China. The disease poses a serious threat to the livelihood of sheep and goat farmers in these regions, thus its control and eradication is a priority in order to ease poverty. The causal agent of PPR is the peste-des-petits ruminants virus (PPRV), which along with rinderpest virus (RPV), measles virus (MeV) and canine distemper virus (CDV) is a member of the Morbillivirus genus within the Paramyxovirus family. These viruses have single stranded negative sense RNA genomes, which contain six transcrip- tion units encoding eight proteins. The nucleocapsid protein (N), phosphoprotein (P) and large polymerase (L) form the minimal replicative unit of the virus, along with the negative sense RNA, and are required for both transcription and replication of the viral RNA. The fusion (F) and hemagglutinin (H) proteins are embedded in the host cell-derived envelope and are required for attachment to cellular receptors and fusion of the viral envelope with host cell membranes. The matrix (M) protein is found inside the virion and ∗ Corresponding author. Tel.: +44 1483231066; fax: +44 1483 231448. E-mail address: Louisa.Chard@bbsrc.ac.uk (L.S. Chard). is believed to be vital for virus particle assembly. The P transcrip- tion unit also encodes two non-structural proteins, C and V. These two proteins are thought to be involved in viral evasion of the host immune responses, specifically by blocking interferon production during infection (Nanda and Baron, 2006). Currently four lineages of PPRV have been identified and, although these do not represent distinct serotypes, they do correspond with geographical distribu- tion of the virus. Here we present the full genome sequences of two virulent strains of PPRV, the Cˆ ote d’Ivoire 1989 strain (CI/89) (lin- eage 2) and the Nigeria 1976 strain (Ng76/1) (lineage 1), the latter being closely related to the Nigeria 1975 (Ng75/1) vaccine strain of PPRV. CI/89 virus was initially isolated from an infected goat from the Cˆ ote d’Ivoire and was kindly provided to us by Dr. A. Diallo (IAEA laboratory, Vienna). This virus was passaged once in our laboratory in primary lamb kidney cells. Ng76/1 was passaged six times on bovine kidney cells and four times on vero cells before being used to infect a flask of Chinese hamster ovary (CHO) cells. Total cellu- lar RNA was extracted from the cells using the Trizol ® technique CI/89 or Ng76/1 cDNA fragments were synthesised from total RNA using Superscript II reverse transcriptase (Invitrogen). PCR reac- tions were performed using the KOD hi-fidelity DNA polymerase (Novagen) using PCR primers based on an available PPRV sequence of the lineage 4 Turkey 2000 (Tu/00) strain (Bailey et al., 2005). The genome promoter and antigenome promoter regions were gener- ated using the RACE technique (rapid amplification of cDNA ends) 0168-1702/$ – see front matter © 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.virusres.2008.04.018