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.
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.
Minet
C
C
CIRAD, UMR CIRAD/INRA Contrôle des Maladies, Montpellier, France.
Yami
M
M
Egzabhier
B
B
Gil
P
P
Tangy
F
F
Brémont
M
M
Libeau
G
G
Diallo
A
A
Albina
E
E
D000595:Amino Acid Sequence; D000818:Animals; D002522:Chlorocebus aethiops; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis / Q000235:genetics; D005814:Genes, Viral; D016679:Genome, Viral; D008969:Molecular Sequence Data; D029021:Peste-des-Petits-Ruminants / Q000378:metabolism / Q000821:virology*; D018107:Peste-des-petits-ruminants virus / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D016415:Sequence Alignment; D017423:Sequence Analysis, RNA; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics*
D012367:RNA, Viral; D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases