ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (2): 213-220.doi:

• 预防兽医 • Previous Articles     Next Articles

Construction and Identification of the Full-length Genomic cDNA Clone of FMDV China99/S Isolated from Swine

CHANG Yan-yan1，ZHENG Hai-xue1，2，JIN Ye1，WANG Guang-xiang1，
SHANG You-jun1，LIU Xiang-tao1*   

  1. 1.Key Laboratory of Animal Virology of Ministry of Agriculture， National FootandMouth Disease Reference Laboratory of China， State Key Laboratory of Veterinary Etiological Biology， Lanzhou Veterinary Research Institute， Chinese Academy of Agricultural Sciences， Lanzhou 730046，China；2. Guangdong Academy of Agricultural Sciences， Guangzhou 510640， China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-02-24 Published:2009-02-24

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Abstract: Total RNA was extracted from tissue samples of FMDV China99/S strain and four overlapping fragments (A， B， C and E) were amplified. The pPⅠcDNA3.1（+）vector with RNA polymerase Ⅰand A， B， C， E fragments were digested respectively by restriction endonucleases(A(BsmBⅠ/XmaⅠ)，B(XmaⅠ/ BamHⅠ)， C(HpaⅠ/KpnⅠ)，E(BamHⅠ/NotⅠ))， and then ligated to produce recombinant plasmid of pPⅠFMDVcDNA3.1（+）. The analysis of nucleotide sequences showed that the genome of swine FMDV China99/S strain is 8 116 nucleotides in length(including Poly(C) and Poly(A) tracts)， and the open reading frame (ORF， 6 318 nucleotides which encode 2 106 amino acids) is flanked by a 5′non-coding region (5′NCR) of 1 064 nucleotides (including 41 C residues Poly(C) tract) and a 3′NCR of 131 nucleotides (including the Poly(A) tail at least 38 A residues). Recombinant plasmid of pPⅠFMDVcDNA3.1（+） was transferred into BHK-21 cells with liposome and typical FMDV pathological changes were observed in subculture. The electron microscope,CPE,animal test,RT-PCR and sequencing analysis verified that FMDV China99/S was rescued.