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《Chinese Journal of Virology》 2010-04
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Construction and Sequencing of Full-length cDNA of Peste des Petits Ruminants Virus

ZHAI Jun-jun,DOU Yong-xi,ZHANG Hai-rui,MAO Li,MENG Xue-lian,LUO Xuo-nong,CAI Xue-peng(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)  
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 75/1strain lays the basis rescuing PPRV effectively and enables further research of PPRV at molecular level.
【Fund】: 农业公益性行业科研专项(200803018);; 甘肃省国际合作项目(0804WCGA133)
【CateGory Index】: S852.659.5
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