title: [Construction and sequencing of full-length cDNA of peste des petits ruminants virus].
abstract: To develop a reverse genetics system of Peste des petits ruminants virus(PPRV), five pairs of oligonucleotide primers were designed on the basis of the full-length genomic sequence of PPRV Nigeria 75/ 1 strain. Using RT-PCR technique, five over-lapping cDNA fragments, designated as JF1, JF2, JF3, JF4 and JF5, respectively, were amplified, followed by cloning into pcDNA3.1(+)vector. An AscI restriction enzyme site and a T7 promoter sequence were introduced immediately upstream of 5'-end, while a PacI restriction enzyme site was engineered downstream of 3'-end. Using pok12 as a plasmid vector, the full-length cDNA clone pok12-PPRV of Nigeria 75/1 was assembled by connecting the five cDNA fragments via the unique restriction endonuclease site of PPRV genome. The resultant nucleotide sequence of the PPRV Nigeria 75/1 strain in the study was compared with other members of genus morbillivirus, and phylogenetic analysis was used to examine the evolutionary relationships. The results showed that PPRV Nigeria 75/ 1 was antigenically closely related to Rinderpest virus and Measles virus. Successful construction of full-length cDNA clone of PPRV Nigeria 75/1 strain lays the basis rescuing PPRV effectively and enables further research of PPRV at molecular level.
authors.affiliation: Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Gansu 730046, China.
authors.forename: Jun-Jun
authors.initials: JJ
authors.lastname: Zhai
authors.forename: Yong-Xi
authors.initials: YX
authors.lastname: Dou
authors.forename: Hai-Rui
authors.initials: HR
authors.lastname: Zhang
authors.forename: Li
authors.initials: L
authors.lastname: Mao
authors.forename: Xue-Lian
authors.initials: XL
authors.lastname: Meng
authors.forename: Xuo-Nong
authors.initials: XN
authors.lastname: Luo
authors.forename: Xue-Peng
authors.initials: XP
authors.lastname: Cai
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D016679:Genome, Viral; D008969:Molecular Sequence Data; D029021:Peste-des-Petits-Ruminants / Q000821:virology; D018107:Peste-des-petits-ruminants virus / Q000145:classification / Q000235:genetics*; D010802:Phylogeny; D017422:Sequence Analysis, DNA
chemical_list: D018076:DNA, Complementary